Identification and expression analyses of two genes encoding putative low-affinity nitrate transporters from Nicotiana plumbaginifolia.

PubMed

Fraisier, V; Dorbe, M F; Daniel-Vedele, F

2001-01-01

Higher plants have both high- and low-affinity nitrate uptake systems (HATS and LATS respectively). Here we report the isolation and characterization of two genes, NpNRT1.1 and NpNRT1.2, from Nicotiana plumbaginifolia whose structural features suggest that they both belong to the NRT1 gene family, which is involved in the LATS. Amino acid sequence alignment showed that the N. plumbaginifolia proteins have greater similarity to their corresponding tomato homologues than to each other. Genomic Southern blot analysis indicates that there are probably more than two members of this family in N. plumbaginifolia. Northern blot analysis shows that NpNRT1.2 expression is restricted strictly to roots, whereas NpNRT1.1, in addition to roots, is expressed at a basal level in all other plant organs. Likewise, differential expression in response to external treatments with various N sources was observed for these two genes: NpNRT1.1 can be considered as a constitutively expressed gene whereas NpNRT1.2 expression is dependent strictly on high nitrate concentrations. Finally, over-expression of a gene involved in the HATS does not lead to any modification of LATS gene expression.

The Nitrate Transporter MtNPF6.8 (MtNRT1.3) Transports Abscisic Acid and Mediates Nitrate Regulation of Primary Root Growth in Medicago truncatula1[W

PubMed Central

Pellizzaro, Anthoni; Clochard, Thibault; Cukier, Caroline; Bourdin, Céline; Juchaux, Marjorie; Montrichard, Françoise; Thany, Steeve; Raymond, Valérie; Planchet, Elisabeth; Morère-Le Paven, Marie-Christine

2014-01-01

Elongation of the primary root during postgermination of Medicago truncatula seedlings is a multigenic trait that is responsive to exogenous nitrate. A quantitative genetic approach suggested the involvement of the nitrate transporter MtNPF6.8 (for Medicago truncatula NITRATE TRANSPORTER1/PEPTIDE TRANSPORTER Family6.8) in the inhibition of primary root elongation by high exogenous nitrate. In this study, the inhibitory effect of nitrate on primary root elongation, via inhibition of elongation of root cortical cells, was abolished in npf6.8 knockdown lines. Accordingly, we propose that MtNPF6.8 mediates nitrate inhibitory effects on primary root growth in M. truncatula. pMtNPF6.8:GUS promoter-reporter gene fusion in Agrobacterium rhizogenes-generated transgenic roots showed the expression of MtNPF6.8 in the pericycle region of primary roots and lateral roots, and in lateral root primordia and tips. MtNPF6.8 expression was insensitive to auxin and was stimulated by abscisic acid (ABA), which restored the inhibitory effect of nitrate in npf6.8 knockdown lines. It is then proposed that ABA acts downstream of MtNPF6.8 in this nitrate signaling pathway. Furthermore, MtNPF6.8 was shown to transport ABA in Xenopus spp. oocytes, suggesting an additional role of MtNPF6.8 in ABA root-to-shoot translocation. 15NO3−-influx experiments showed that only the inducible component of the low-affinity transport system was affected in npf6.8 knockdown lines. This indicates that MtNPF6.8 is a major contributor to the inducible component of the low-affinity transport system. The short-term induction by nitrate of the expression of Nitrate Reductase1 (NR1) and NR2 (genes that encode two nitrate reductase isoforms) was greatly reduced in the npf6.8 knockdown lines, supporting a role of MtNPF6.8 in the primary nitrate response in M. truncatula. PMID:25367858

Toxic effects of lead and nickel nitrate on rat liver chromatin components.

PubMed

Rabbani-Chadegani Iii, Azra; Fani, Nesa; Abdossamadi, Sayeh; Shahmir, Nosrat

2011-01-01

The biological activity of heavy metals is related to their physicochemical interaction with biological receptors. In the present study, the effect of low concentrations of nickel nitrate and lead nitrate (<0.3 mM) on rat liver soluble chromatin and histone proteins was examined. The results showed that addition of various concentrations of metals to chromatin solution preceded the chromatin into aggregation and precipitation in a dose-dependant manner; however, the extent of absorbance changes at 260 and 400 nm was different between two metals. Gel electrophoresis of histone proteins and DNA of the supernatants obtained from the metal-treated chromatin and the controls revealed higher affinity of lead nitrate to chromatin compared to nickel nitrate. Also, the binding affinity of lead nitrate to histone proteins free in solution was higher than nickel. On the basis of the results, it is concluded that lead reacts with chromatin components even at very low concentrations and induce chromatin aggregation through histone-DNA cross-links. Whereas, nickel nitrate is less effective on chromatin at low concentrations, suggesting higher toxicity of lead nitrate on chromatin compared to nickel. Copyright © 2010 Wiley Periodicals, Inc.

Kinetic analysis of a complete nitrifier reveals an oligotrophic lifestyle

PubMed Central

Kits, K. Dimitri; Sedlacek, Christopher J.; Lebedeva, Elena V.; Han, Ping; Bulaev, Alexandr; Pjevac, Petra; Daebeler, Anne; Romano, Stefano; Albertsen, Mads; Stein, Lisa Y.; Daims, Holger; Wagner, Michael

2017-01-01

Summary paragraph Nitrification, the oxidation of ammonia (NH3) via nitrite (NO2-) to nitrate (NO3-), is a key process of the biogeochemical nitrogen cycle. For decades, ammonia and nitrite oxidation were thought to be separately catalyzed by ammonia-oxidizing bacteria (AOB) and archaea (AOA), and by nitrite-oxidizing bacteria (NOB). The recent discovery of complete ammonia oxidizers (comammox) in the NOB genus Nitrospira1,2, which alone convert ammonia to nitrate, raised questions about the ecological niches where comammox Nitrospira successfully compete with canonical nitrifiers. Here we isolated the first pure culture of a comammox bacterium, Nitrospira inopinata, and show that it is adapted to slow growth in oligotrophic and dynamic habitats based on a high affinity for ammonia, low maximum rate of ammonia oxidation, high growth yield compared to canonical nitrifiers, and genomic potential for alternative metabolisms. The nitrification kinetics of four AOA from soil and hot springs were determined for comparison. Their surprisingly poor substrate affinities and lower growth yields reveal that, in contrast to earlier assumptions, not all AOA are most competitive in strongly oligotrophic environments and that N. inopinata has the highest substrate affinity of all analyzed ammonia oxidizer isolates except the marine AOA Nitrosopumilus maritimus SCM13. These results suggest a role of comammox organisms for nitrification under oligotrophic and dynamic conditions. PMID:28847001

Glucose Elevates NITRATE TRANSPORTER2.1 Protein Levels and Nitrate Transport Activity Independently of Its HEXOKINASE1-Mediated Stimulation of NITRATE TRANSPORTER2.1 Expression1[W][OPEN

PubMed Central

de Jong, Femke; Thodey, Kate; Lejay, Laurence V.; Bevan, Michael W.

2014-01-01

Mineral nutrient uptake and assimilation is closely coordinated with the production of photosynthate to supply nutrients for growth. In Arabidopsis (Arabidopsis thaliana), nitrate uptake from the soil is mediated by genes encoding high- and low-affinity transporters that are transcriptionally regulated by both nitrate and photosynthate availability. In this study, we have studied the interactions of nitrate and glucose (Glc) on gene expression, nitrate transport, and growth using glucose-insensitive2-1 (gin2-1), which is defective in sugar responses. We confirm and extend previous work by showing that HEXOKINASE1-mediated oxidative pentose phosphate pathway (OPPP) metabolism is required for Glc-mediated NITRATE TRANSPORTER2.1 (NRT2.1) expression. Treatment with pyruvate and shikimate, two products derived from intermediates of the OPPP that are destined for amino acid production, restores wild-type levels of NRT2.1 expression, suggesting that metabolites derived from OPPP metabolism can, together with Glc, directly stimulate high levels of NRT2.1 expression. Nitrate-mediated NRT2.1 expression is not influenced by gin2-1, showing that Glc does not influence NRT2.1 expression through nitrate-mediated mechanisms. We also show that Glc stimulates NRT2.1 protein levels and transport activity independently of its HEXOKINASE1-mediated stimulation of NRT2.1 expression, demonstrating another possible posttranscriptional mechanism influencing nitrate uptake. In gin2-1 plants, nitrate-responsive biomass growth was strongly reduced, showing that the supply of OPPP metabolites is essential for assimilating nitrate for growth. PMID:24272701

Genes responding to water deficit in apple (Malus × domestica Borkh.) roots.

PubMed

Bassett, Carole Leavel; Baldo, Angela M; Moore, Jacob T; Jenkins, Ryan M; Soffe, Doug S; Wisniewski, Michael E; Norelli, John L; Farrell, Robert E

2014-07-08

Individual plants adapt to their immediate environment using a combination of biochemical, morphological and life cycle strategies. Because woody plants are long-lived perennials, they cannot rely on annual life cycle strategies alone to survive abiotic stresses. In this study we used suppression subtractive hybridization to identify genes both up- and down-regulated in roots during water deficit treatment and recovery. In addition we followed the expression of select genes in the roots, leaves, bark and xylem of 'Royal Gala' apple subjected to a simulated drought and subsequent recovery. In agreement with studies from both herbaceous and woody plants, a number of common drought-responsive genes were identified, as well as a few not previously reported. Three genes were selected for more in depth analysis: a high affinity nitrate transporter (MdNRT2.4), a mitochondrial outer membrane translocase (MdTOM7.1), and a gene encoding an NPR1 homolog (MpNPR1-2). Quantitative expression of these genes in apple roots, bark and leaves was consistent with their roles in nutrition and defense. Additional genes from apple roots responding to drought were identified using suppression subtraction hybridization compared to a previous EST analysis from the same organ. Genes up- and down-regulated during drought recovery in roots were also identified. Elevated levels of a high affinity nitrate transporter were found in roots suggesting that nitrogen uptake shifted from low affinity transport due to the predicted reduction in nitrate concentration in drought-treated roots. Suppression of a NPR1 gene in leaves of drought-treated apple trees may explain in part the increased disease susceptibility of trees subjected to dehydrative conditions.

High-affinity nitrate/nitrite transporters NrtA and NrtB of Aspergillus nidulans exhibit high specificity and different inhibitor sensitivity

PubMed Central

Akhtar, Naureen; Karabika, Eugenia; Kinghorn, James R.; Glass, Anthony D.M.; Unkles, Shiela E.

2015-01-01

The NrtA and NrtB nitrate transporters are paralogous members of the major facilitator superfamily in Aspergillus nidulans. The availability of loss-of-function mutations allowed individual investigation of the specificity and inhibitor sensitivity of both NrtA and NrtB. In this study, growth response tests were carried out at a growth-limiting concentration of nitrate (1 mM) as the sole nitrogen source, in the presence of a number of potential nitrate analogues at various concentrations, to evaluate their effect on nitrate transport. Both chlorate and chlorite inhibited fungal growth, with chlorite exerting the greater inhibition. The main transporter of nitrate, NrtA, proved to be more sensitive to chlorate than the minor transporter, NrtB. Similarly, the cation caesium was shown to exert differential effects, strongly inhibiting the activity of NrtB, but not NrtA. In contrast, no inhibition of nitrate uptake by NrtA or NrtB transporters was observed in either growth tests or uptake assays in the presence of bicarbonate, formate, malonate or oxalate (sulphite could not be tested in uptake assays owing to its reaction with nitrate), indicating significant specificity of nitrate transport. Kinetic analyses of nitrate uptake revealed that both chlorate and chlorite inhibited NrtA competitively, while these same inhibitors inhibited NrtB in a non-competitive fashion. The caesium ion appeared to inhibit NrtA in a non-competitive fashion, while NrtB was inhibited uncompetitively. The results provide further evidence of the distinctly different characteristics as well as the high specificity of nitrate uptake by these two transporters. PMID:25855763

Disruption of the rice nitrate transporter OsNPF2.2 hinders root-to-shoot nitrate transport and vascular development

PubMed Central

Li, Yuge; Ouyang, Jie; Wang, Ya-Yun; Hu, Rui; Xia, Kuaifei; Duan, Jun; Wang, Yaqin; Tsay, Yi-Fang; Zhang, Mingyong

2015-01-01

Plants have evolved to express some members of the nitrate transporter 1/peptide transporter family (NPF) to uptake and transport nitrate. However, little is known of the physiological and functional roles of this family in rice (Oryza sativa L.). Here, we characterized the vascular specific transporter OsNPF2.2. Functional analysis using cDNA-injected Xenopus laevis oocytes revealed that OsNPF2.2 is a low-affinity, pH-dependent nitrate transporter. Use of a green fluorescent protein tagged OsNPF2.2 showed that the transporter is located in the plasma membrane in the rice protoplast. Expression analysis showed that OsNPF2.2 is nitrate inducible and is mainly expressed in parenchyma cells around the xylem. Disruption of OsNPF2.2 increased nitrate concentration in the shoot xylem exudate when nitrate was supplied after a deprivation period; this result suggests that OsNPF2.2 may participate in unloading nitrate from the xylem. Under steady-state nitrate supply, the osnpf2.2 mutants maintained high levels of nitrate in the roots and low shoot:root nitrate ratios; this observation suggests that OsNPF2.2 is involved in root-to-shoot nitrate transport. Mutation of OsNPF2.2 also caused abnormal vasculature and retarded plant growth and development. Our findings demonstrate that OsNPF2.2 can unload nitrate from the xylem to affect the root-to-shoot nitrate transport and plant development. PMID:25923512

Constitutive expression of a putative high-affinity nitrate transporter in Nicotiana plumbaginifolia: evidence for post-transcriptional regulation by a reduced nitrogen source.

PubMed

Fraisier, V; Gojon, A; Tillard, P; Daniel-Vedele, F

2000-08-01

The NpNRT2.1 gene encodes a putative inducible component of the high-affinity nitrate (NO3-) uptake system in Nicotiana plumbaginifolia. Here we report functional and physiological analyses of transgenic plants expressing the NpNRT2.1 coding sequence fused to the CaMV 35S or rolD promoters. Irrespective of the level of NO3- supplied, NO3- contents were found to be remarkably similar in wild-type and transgenic plants. Under specific conditions (growth on 10 mM NO3-), the steady-state NpNRT2. 1 mRNA level resulting from the deregulated transgene expression was accompanied by an increase in 15NO3- influx measured in the low concentration range. This demonstrates for the first time that the NRT2.1 sequence codes a limiting element of the inducible high-affinity transport system. Both 15NO3- influx and mRNA levels decreased in the wild type after exposure to ammonium, in agreement with previous results from many species. Surprisingly, however, influx was also markedly decreased in transgenic plants, despite stable levels of transgene expression in independent transformants after ammonium addition. We conclude that the conditions associated with the supply of a reduced nitrogen source such as ammonium, or with the generation of a further downstream metabolite, probably exert a repressive effect on NO3- influx at both transcriptional and post-transcriptional levels.

Two phloem nitrate transporters, NRT1.11 and NRT1.12, are important for redistributing xylem-borne nitrate to enhance plant growth.

PubMed

Hsu, Po-Kai; Tsay, Yi-Fang

2013-10-01

This study of the Arabidopsis (Arabidopsis thaliana) nitrate transporters NRT1.11 and NRT1.12 reveals how the interplay between xylem and phloem transport of nitrate ensures optimal nitrate distribution in leaves for plant growth. Functional analysis in Xenopus laevis oocytes showed that both NRT1.11 and NRT1.12 are low-affinity nitrate transporters. Quantitative reverse transcription-polymerase chain reaction and immunoblot analysis showed higher expression of these two genes in larger expanded leaves. Green fluorescent protein and β-glucuronidase reporter analyses indicated that NRT1.11 and NRT1.12 are plasma membrane transporters expressed in the companion cells of the major vein. In nrt1.11 nrt1.12 double mutants, more root-fed (15)NO3(-) was translocated to mature and larger expanded leaves but less to the youngest tissues, suggesting that NRT1.11 and NRT1.12 are required for transferring root-derived nitrate into phloem in the major veins of mature and larger expanded leaves for redistributing to the youngest tissues. Distinct from the wild type, nrt1.11 nrt1.12 double mutants show no increase of plant growth at high nitrate supply. These data suggested that NRT1.11 and NRT1.12 are involved in xylem-to-phloem transfer for redistributing nitrate into developing leaves, and such nitrate redistribution is a critical step for optimal plant growth enhanced by increasing external nitrate.

Nitrate supplementation and human exercise performance: too much of a good thing?

PubMed

Poortmans, Jacques R; Gualano, Bruno; Carpentier, Alain

2015-11-01

Ergogenic supplements in sport events are widely used by popular and competitive athletes to enhance performance and reduce oxygen cost. Beetroot juice and nitrate salts have been increasingly used for the past 5-6 years. The present review discusses the scientific background, the efficiency and potential adverse effects of excessive nitrate supplementation. There is clear evidence that nitrate from different food ingredients (such as beetroot juice and other vegetables) is converted into nitrite and possibly into nitric oxide, which may promote vasodilation, angiogenesis and mitochondrial biogenesis. The high affinity of nitric oxide towards different enzyme pathways inhibits excessive mitochondrial respiration and, therefore, tissue oxygen consumption. In addition, L-arginine supplements are proposed to stimulate nitric oxide synthesis in the endothelium. On the basis of these biochemical properties, nitrate supplementation has been suggested to athletes to enhance exercise performance. The recent publications in human individuals based on L-arginine, beetroot juice or nitrate supplementation revealed either a minor positive effect or no systematic effect on exercise performance, especially in trained athletes. Of note, the sugar content of whole beetroot juice might induce a slightly more pronounced effect. Although reasonable intake of nitrate salts (up to 1 g/day) has no detrimental effect on kidney function, the risk and benefit of higher nitrate intake needs to be evaluated to define the optimal range of supplementation.

Effects of Atrazine, Metolachlor, Carbaryl and Chlorothalonil on Benthic Microbes and Their Nutrient Dynamics

PubMed Central

Elias, Daniel; Bernot, Melody J.

2014-01-01

Atrazine, metolachlor, carbaryl, and chlorothalonil are detected in streams throughout the U.S. at concentrations that may have adverse effects on benthic microbes. Sediment samples were exposed to these pesticides to quantify responses of ammonium, nitrate, and phosphate uptake by the benthic microbial community. Control uptake rates of sediments had net remineralization of nitrate (−1.58 NO3 µg gdm−1 h−1), and net assimilation of phosphate (1.34 PO4 µg gdm−1 h−1) and ammonium (0.03 NH4 µg gdm−1 h−1). Metolachlor decreased ammonium and phosphate uptake. Chlorothalonil decreased nitrate remineralization and phosphate uptake. Nitrate, ammonium, and phosphate uptake rates are more pronounced in the presence of these pesticides due to microbial adaptations to toxicants. Our interpretation of pesticide availability based on their water/solid affinities supports no effects for atrazine and carbaryl, decreasing nitrate remineralization, and phosphate assimilation in response to chlorothalonil. Further, decreased ammonium and phosphate uptake in response to metolachlor is likely due to affinity. Because atrazine target autotrophs, and carbaryl synaptic activity, effects on benthic microbes were not hypothesized, consistent with results. Metolachlor and chlorothalonil (non-specific modes of action) had significant effects on sediment microbial nutrient dynamics. Thus, pesticides with a higher affinity to sediments and/or broad modes of action are likely to affect sediment microbes' nutrient dynamics than pesticides dissolved in water or specific modes of action. Predicted nutrient uptake rates were calculated at mean and peak concentrations of metolachlor and chlorothalonil in freshwaters using polynomial equations generated in this experiment. We concluded that in natural ecosystems, peak chlorothalonil and metolachlor concentrations could affect phosphate and ammonium by decreasing net assimilation, and nitrate uptake rates by decreasing remineralization, relative to mean concentrations of metolachlor and chlorothalonil. Our regression equations can complement models of nitrogen and phosphorus availability in streams to predict potential changes in nutrient dynamics in response to pesticides in freshwaters. PMID:25275369

Characterization of Tyrosine Nitration and Cysteine Nitrosylation Modifications by Metastable Atom-Activation Dissociation Mass Spectrometry

NASA Astrophysics Data System (ADS)

Cook, Shannon L.; Jackson, Glen P.

2011-02-01

The fragmentation behavior of nitrated and S-nitrosylated peptides were studied using collision induced dissociation (CID) and metastable atom-activated dissociation mass spectrometry (MAD-MS). Various charge states, such as 1+, 2+, 3+, 2-, of modified and unmodified peptides were exposed to a beam of high kinetic energy helium (He) metastable atoms resulting in extensive backbone fragmentation with significant retention of the post-translation modifications (PTMs). Whereas the high electron affinity of the nitrotyrosine moiety quenches radical chemistry and fragmentation in electron capture dissociation (ECD) and electron transfer dissociation (ETD), MAD does produce numerous backbone cleavages in the vicinity of the modification. Fragment ions of nitrosylated cysteine modifications typically exhibit more abundant neutral losses than nitrated tyrosine modifications because of the extremely labile nature of the nitrosylated cysteine residues. However, compared with CID, MAD produced between 66% and 86% more fragment ions, which preserved the labile -NO modification. MAD was also able to differentiate I/L residues in the modified peptides. MAD is able to induce radical ion chemistry even in the presence of strong radical traps and therefore offers unique advantages to ECD, ETD, and CID for determination of PTMs such as nitrated and S-nitrosylated peptides.

Evaluation of the plant growth-promoting activity of Pseudomonas nitroreducens in Arabidopsis thaliana and Lactuca sativa.

PubMed

Trinh, Cao Son; Lee, Hyeri; Lee, Won Je; Lee, Seok Jin; Chung, Namhyun; Han, Juhyeong; Kim, Jongyun; Hong, Suk-Whan; Lee, Hojoung

2018-06-01

Pseudomonas nitroreducens: strain IHB B 13561 (PnIHB) enhances the growth of Arabidopsis thaliana and Lactuca sativa via the stimulation of cell development and nitrate absorption. Plant growth-promoting rhizobacteria (PGPR) enhance plant development through various mechanisms; they improve the uptake of soil resources by plants to greatly promote plant growth. Here, we used Arabidopsis thaliana seedlings and Lactuca sativa to screen the growth enhancement activities of a purified PGPR, Pseudomonas nitroreducens strain IHB B 13561 (PnIHB). When cocultivated with PnIHB, both species of plants exhibited notably improved growth, particularly in regard to biomass. Quantitative reverse transcription polymerase chain reaction analysis indicated high expression levels of the nitrate transporter genes, especially NRT2.1, which plays a major role in the high-affinity nitrate transport system in roots. Moreover, enhanced activity of the cyclin-B1 promoter was observed when wild-type 'Columbia-0' Arabidopsis seedlings were exposed to PnIHB, whereas upregulation of cyclin-B also occurred in the inoculated lettuce seedlings. Overall, these results suggest that PnIHB improves A. thaliana and L. sativa growth via specific pathways involved in the promotion of cell development and enhancement of nitrate uptake.

Biotic and environmental stress induces nitration and changes in structure and function of the sea urchin major yolk protein toposome.

PubMed

Castellano, Immacolata; Migliaccio, Oriana; Ferraro, Giarita; Maffioli, Elisa; Marasco, Daniela; Merlino, Antonello; Zingone, Adriana; Tedeschi, Gabriella; Palumbo, Anna

2018-03-15

The major yolk protein toposome plays crucial roles during gametogenesis and development of sea urchins. We previously found that nitration of toposome increases in the gonads of a Paracentrotus lividus population living in a marine protected area affected by toxic blooms of Ostreospsis cf. ovata, compared to control populations. This modification is associated with ovatoxin accumulation, high levels of nitric oxide in the gonads, and a remarkable impairment of progeny development. However, nothing is known about the environmental-mediated-regulation of the structure and biological function of toposome. Here, we characterize through wide-ranging biochemical and structural analyses the nitrated toposome of sea urchins exposed to the bloom, and subsequently detoxified. The increased number of nitrated tyrosines in toposome of sea urchins collected during algal bloom induced structural changes and improvement of the Ca 2+ -binding affinity of the protein. After 3 months' detoxification, ovatoxin was undetectable, and the number of nitric oxide-modified tyrosines was reduced. However, the nitration of specific residues was irreversible and occurred also in embryos treated with metals, used as a proxy of environmental pollutants. The structural and functional changes of toposome caused by nitration under adverse environmental conditions may be related to the defective development of sea urchins' progeny.

Composting-derived organic coating on biochar enhances its affinity to nitrate

NASA Astrophysics Data System (ADS)

Hagemann, Nikolas; Joseph, Stephen; Conte, Pellegrino; Albu, Mihaela; Obst, Martin; Borch, Thomas; Orsetti, Silvia; Subdiaga, Edisson; Behrens, Sebastian; Kappler, Andreas

2017-04-01

Biochar is defined charcoal that is produced by the thermal treatment of biomass in the (partial) absence of oxygen (pyrolysis) for non-oxidative applications, especially in agriculture. Due to its high surface area and porous structure, it is suggested as a beneficial soil amendment to increase crop yields and to tailor biogeochemical cycles in agro-ecosystems to reduce both greenhouse gas emissions and nutrient leaching. While early research focused on single applications of large amounts of biochar (>10 t ha-1), economic and ecological boundaries as well as practical considerations and recent findings shifted the focus towards low-dose (˜1 t ha-1) and potentially repeated applications of nutrient-enriched biochars, i.e. biochar-based fertilizers in the root-zone. Thus, biochar must be "loaded" with nutrients prior to its use as a root-zone amendment. Co-composting is suggested as a superior method, as co-composted biochar was shown to promote plant growth and showed the desired slow release of nutrients such as nitrate ("nitrate capture", Kammann et al., 2015 SR5:11080). However, the underlying mechanisms are not understood and nitrate capture has been quantified only for isolated biochars but not for e.g. biochar-amended composts without prior separation of the biochar. In the present study, we used repeated extractions with 2 M KCl and found that up to 30% of the nitrate present in a biochar-amended compost is captured in biochar, although biochar was amended to the initial composting feedstock (manure) only at 4% (w/w). Additionally, we quantified nitrate capture by pristine biochar after soaking the biochar in NH4NO3 solution in the absence of any additional organic carbon and nitrate capture of separated co-composted biochar. Assuming pseudo-first order kinetics for biochar nitrate release, we found an increase of biochar's affinity to nitrate after co-composting. Spectro-microscopical investigations (scanning transmission electron microscopy with electron energy loss spectroscopy - STEM-EELS, scanning transmission X-ray microscopy STXM) revealed the formation of a nano-porous organic coating on co-composted biochar. This coating alters the interaction of biochar with water as evidenced by proton fast field cycling nuclear magnetic resonance (1H FFC NMR) relaxometry and might explain its distinct characteristics. Our findings offer a roadmap for future research to design sustainable slow-release nitrogen fertilizers based on biochar to reduce the environmental impact of agriculture. Further microscopic studies are necessary to understand the preconditions of the formation of organic coatings on biochar on a holistic basis to design biochar post-production treatments.

Knockdown of a Rice Stelar Nitrate Transporter Alters Long-Distance Translocation But Not Root Influx1[W][OA

PubMed Central

Tang, Zhong; Fan, Xiaorong; Li, Qing; Feng, Huimin; Miller, Anthony J.; Shen, Qirong; Xu, Guohua

2012-01-01

Root nitrate uptake is well known to adjust to the plant’s nitrogen demand for growth. Long-distance transport and/or root storage pools are thought to provide negative feedback signals regulating root uptake. We have characterized a vascular specific nitrate transporter belonging to the high-affinity Nitrate Transporter2 (NRT2) family, OsNRT2.3a, in rice (Oryza sativa ssp. japonica ‘Nipponbare’). Localization analyses using protoplast expression, in planta promoter-β-glucuronidase assay, and in situ hybridization showed that OsNRT2.3a was located in the plasma membrane and mainly expressed in xylem parenchyma cells of the stele of nitrate-supplied roots. Knockdown expression of OsNRT2.3a by RNA interference (RNAi) had impaired xylem loading of nitrate and decreased plant growth at low (0.5 mm) nitrate supply. In comparison with the wild type, the RNAi lines contained both nitrate and total nitrogen significantly higher in the roots and lower in the shoots. The short-term [15N]NO3− influx (5 min) in entire roots and NO3− fluxes in root surfaces showed that the knockdown of OsNRT2.3a in comparison with the wild type did not affect nitrate uptake by roots. The RNAi plants showed no significant changes in the expression of some root nitrate transporters (OsNRT2.3b, OsNRT2.4, and OsNAR2.1), but transcripts for nia1 (nitrate reductase) had increased and OsNRT2.1 and OsNRT2.2 had decreased when the plants were supplied with nitrate. Taken together, the data demonstrate that OsNRT2.3a plays a key role in long-distance nitrate transport from root to shoot at low nitrate supply level in rice. PMID:23093362

Dual regulation of root hydraulic conductivity and plasma membrane aquaporins by plant nitrate accumulation and high-affinity nitrate transporter NRT2.1.

PubMed

Li, Guowei; Tillard, Pascal; Gojon, Alain; Maurel, Christophe

2016-04-01

The water status and mineral nutrition of plants critically determine their growth and development. Nitrate (NO3(-)), the primary nitrogen source of higher plants, is known to impact the water transport capacity of roots (root hydraulic conductivity, Lpr). To explore the effects and mode of action of NO3(-) on Lpr, we used an extended set of NO3(-) transport (nrt1.1, nrt1.2, nrt1.5 and nrt2.1), signaling (nrt1.1 and nrt2.1) and metabolism (nia) mutants in Arabidopsis, grown under various NO3(-) conditions. First, a strong positive relationship between Lpr and NO3(-) accumulation, in shoots rather than in roots, was revealed. Secondly, a specific 30% reduction of Lpr in nrt2.1 plants unraveled a major role for the high-affinity NO3(-) transporter NRT2.1 in increasing Lpr These results indicate that NO3(-)signaling rather than nitrogen assimilation products governs Lpr in Arabidopsis. Quantitative real-time reverse transcription-PCR and enzyme-linked immunosorbent assays (ELISAs) were used to investigate the effects of NO3(-) availability on plasma membrane aquaporin (plasma membrane intrinsic protein; PIP) expression. Whereas PIP regulation mostly occurs at the post-translational level in wild-type plants, a regulation of PIPs at both the transcriptional and translational levels was uncovered in nrt2.1 plants. In conclusion, this work reveals that control of Arabidopsis Lpr and PIP functions by NO3(-) involves novel shoot to root signaling and NRT2.1-dependent functions. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Purification of nitrate reductase from Nicotiana plumbaginifolia by affinity chromatography using 5'AMP-sepharose and monoclonal antibodies.

PubMed

Moureaux, T; Leydecker, M T; Meyer, C

1989-02-15

Nitrate reductase was purified from leaves of Nicotiana plumbaginifolia using either 5'AMP-Sepharose chromatography or two steps of immunoaffinity chromatography involving monoclonal antibodies directed against nitrate reductase from maize and against ribulose-1,5-bisphosphate carboxylase from N. plumbaginifolia. Nitrate reductase obtained by the first method was purified 1000-fold to a specific activity of 9 units/mg protein. The second method produced an homogenous enzyme, purified 21,000-fold to a specific activity of 80 units/mg protein. SDS/PAGE of nitrate reductase always resulted in two bands of 107 and 99.5 kDa. The 107-kDa band was the nitrate reductase subunit of N. plumbaginifolia; the smaller one of 99.5 kDa is thought, as commonly reported, to result from proteolysis of the larger protein. The molecular mass of 107 kDa is close to the values calculated from the coding sequences of the two nitrate reductase genes recently cloned from tobacco (Nicotiana tabacum cv Xanthi).

Energy coupling to nitrate uptake into the denitrifying cells of Paracoccus denitrificans.

PubMed

Kucera, Igor

2005-09-05

This study deals with the effects of the agents that dissipate the individual components of the proton motive force (short-chain fatty acids, nigericin, and valinomycin) upon the methyl viologen-coupled nitrate reductase activity in intact cells. Substitution of butyrate or acetate for chloride in Tris-buffered assay media resulted in a marked inhibition at pH 7. In a Tris--chloride buffer of neutral pH, the reaction was almost fully inhibitable by nigericin. Alkalinisation increased the IC(50) value for nigericin and decreased the maximal inhibition attained. Both types of inhibitions could be reversed by the permeabilisation of cells or by the addition of nitrite, and that caused by nigericin disappeared at high extracellular concentrations of potassium. These data indicate that nitrate transport step relies heavily on the pH gradient at neutral pH. Since the affinity of cells for nitrate was strongly diminished by imposing an inside-positive potassium (or lithium) diffusion potential at alkaline external pH, a potential dependent step may be of significance in the transporter cycle under these conditions. Experiments with sodium-depleted media provided no hints for Na(+) as a possible H(+) substitute.

RAPID NITRATE UPTAKE RATES AND LARGE SHORT-TERM STORAGE CAPACITIES MAY EXPLAIN WHY OPPORTUNISTIC GREEN MACROALGAE DOMINATE SHALLOW EUTROPHIC ESTUARIES1.

PubMed

Kennison, Rachel L; Kamer, Krista; Fong, Peggy

2011-06-01

We quantified the effects of initial macroalgal tissue nitrogen (N) status (depleted and enriched) and varying pulses of nitrate (NO 3 - ) concentration on uptake and storage of nitrogen in Ulva intestinalis L. and Ulva expansa (Setch.) Setch. et N. L. Gardner using mesocosms modeling shallow coastal estuaries in Mediterranean climates. Uptake of NO 3 - (μmol · g dry weight [dwt] -1  · h -1 ) was measured as loss from the water after 1, 2, 4, 8, 12, and 24 h and storage as total tissue nitrogen (% dwt) and nitrate (ppm). Both species of algae exhibited a high affinity for NO 3 - across all N pulses and initial tissue contents. There was greater NO 3 - removal from the water for depleted than enriched algae across all time intervals. In the low-N-pulse treatment, U. intestinalis and U. expansa removed all measurable NO 3 - within 8 and 12 h, respectively, and in the medium and high treatments, removal was high and then decreased over time. Maximum mean uptake rates of nitrate were greater for U. expansa (∼300 μmol · g dwt -1  · h -1 ) than U. intestinalis (∼100 μmol · g dwt -1  · h -1 ); however, uptake rates were highly variable over time. Overall, U. expansa uptake rates were double those of U. intestinalis. Maximum tissue NO 3 - for U. expansa was >1,000 ppm, five times that of U. intestinalis, suggesting that U. expansa has a greater storage capacity in this cellular pool. These results showed that opportunistic green algae with differing tissue nutrient histories were able to efficiently remove nitrate from the water across a wide range of N pulses; thus, both are highly adapted to proliferate in estuarine environments with pulsed nutrient supplies. © 2011 Phycological Society of America.

Mechanisms of direct inhibition of the respiratory sulfate-reduction pathway by (per)chlorate and nitrate.

PubMed

Carlson, Hans K; Kuehl, Jennifer V; Hazra, Amrita B; Justice, Nicholas B; Stoeva, Magdalena K; Sczesnak, Andrew; Mullan, Mark R; Iavarone, Anthony T; Engelbrektson, Anna; Price, Morgan N; Deutschbauer, Adam M; Arkin, Adam P; Coates, John D

2015-06-01

We investigated perchlorate (ClO(4)(-)) and chlorate (ClO(3)(-)) (collectively (per)chlorate) in comparison with nitrate as potential inhibitors of sulfide (H(2)S) production by mesophilic sulfate-reducing microorganisms (SRMs). We demonstrate the specificity and potency of (per)chlorate as direct SRM inhibitors in both pure cultures and undefined sulfidogenic communities. We demonstrate that (per)chlorate and nitrate are antagonistic inhibitors and resistance is cross-inducible implying that these compounds share at least one common mechanism of resistance. Using tagged-transposon pools we identified genes responsible for sensitivity and resistance in Desulfovibrio alaskensis G20. We found that mutants in Dde_2702 (Rex), a repressor of the central sulfate-reduction pathway were resistant to both (per)chlorate and nitrate. In general, Rex derepresses its regulon in response to increasing intracellular NADH:NAD(+) ratios. In cells in which respiratory sulfate reduction is inhibited, NADH:NAD(+) ratios should increase leading to derepression of the sulfate-reduction pathway. In support of this, in (per)chlorate or nitrate-stressed wild-type G20 we observed higher NADH:NAD(+) ratios, increased transcripts and increased peptide counts for genes in the core Rex regulon. We conclude that one mode of (per)chlorate and nitrate toxicity is as direct inhibitors of the central sulfate-reduction pathway. Our results demonstrate that (per)chlorate are more potent inhibitors than nitrate in both pure cultures and communities, implying that they represent an attractive alternative for controlling sulfidogenesis in industrial ecosystems. Of these, perchlorate offers better application logistics because of its inhibitory potency, solubility, relative chemical stability, low affinity for mineral cations and high mobility in environmental systems.

Nitrous oxide production kinetics during nitrate reduction in river sediments.

PubMed

Laverman, Anniet M; Garnier, Josette A; Mounier, Emmanuelle M; Roose-Amsaleg, Céline L

2010-03-01

A significant amount of nitrogen entering river basins is denitrified in riparian zones. The aim of this study was to evaluate the influence of nitrate and carbon concentrations on the kinetic parameters of nitrate reduction as well as nitrous oxide emissions in river sediments in a tributary of the Marne (the Seine basin, France). In order to determine these rates, we used flow-through reactors (FTRs) and slurry incubations; flow-through reactors allow determination of rates on intact sediment slices under controlled conditions compared to sediment homogenization in the often used slurry technique. Maximum nitrate reduction rates (R(m)) ranged between 3.0 and 7.1microg Ng(-1)h(-1), and affinity constant (K(m)) ranged from 7.4 to 30.7mg N-NO(3)(-)L(-1). These values were higher in slurry incubations with an R(m) of 37.9microg Ng(-1)h(-1) and a K(m) of 104mg N-NO(3)(-)L(-1). Nitrous oxide production rates did not follow Michaelis-Menten kinetics, and we deduced a rate constant with an average of 0.7 and 5.4ng Ng(-1)h(-1) for FTR and slurry experiments respectively. The addition of carbon (as acetate) showed that carbon was not limiting nitrate reduction rates in these sediments. Similar rates were obtained for FTR and slurries with carbon addition, confirming the hypothesis that homogenization increases rates due to release of and increasing access to carbon in slurries. Nitrous oxide production rates in FTR with carbon additions were low and represented less than 0.01% of the nitrate reduction rates and were even negligible in slurries. Maximum nitrate reduction rates revealed seasonality with high potential rates in fall and winter and low rates in late spring and summer. Under optimal conditions (anoxia, non-limiting nitrate and carbon), nitrous oxide emission rates were low, but significant (0.01% of the nitrate reduction rates). Copyright 2009 Elsevier Ltd. All rights reserved.

Mutation of the Arabidopsis NRT1.5 nitrate transporter causes defective root-to-shoot nitrate transport.

PubMed

Lin, Shan-Hua; Kuo, Hui-Fen; Canivenc, Geneviève; Lin, Choun-Sea; Lepetit, Marc; Hsu, Po-Kai; Tillard, Pascal; Lin, Huey-Ling; Wang, Ya-Yun; Tsai, Chyn-Bey; Gojon, Alain; Tsay, Yi-Fang

2008-09-01

Little is known about the molecular and regulatory mechanisms of long-distance nitrate transport in higher plants. NRT1.5 is one of the 53 Arabidopsis thaliana nitrate transporter NRT1 (Peptide Transporter PTR) genes, of which two members, NRT1.1 (CHL1 for Chlorate resistant 1) and NRT1.2, have been shown to be involved in nitrate uptake. Functional analysis of cRNA-injected Xenopus laevis oocytes showed that NRT1.5 is a low-affinity, pH-dependent bidirectional nitrate transporter. Subcellular localization in plant protoplasts and in planta promoter-beta-glucuronidase analysis, as well as in situ hybridization, showed that NRT1.5 is located in the plasma membrane and is expressed in root pericycle cells close to the xylem. Knockdown or knockout mutations of NRT1.5 reduced the amount of nitrate transported from the root to the shoot, suggesting that NRT1.5 participates in root xylem loading of nitrate. However, root-to-shoot nitrate transport was not completely eliminated in the NRT1.5 knockout mutant, and reduction of NRT1.5 in the nrt1.1 background did not affect root-to-shoot nitrate transport. These data suggest that, in addition to that involving NRT1.5, another mechanism is responsible for xylem loading of nitrate. Further analyses of the nrt1.5 mutants revealed a regulatory loop between nitrate and potassium at the xylem transport step.

Interaction of Yna1 and Yna2 Is Required for Nuclear Accumulation and Transcriptional Activation of the Nitrate Assimilation Pathway in the Yeast Hansenula polymorpha.

PubMed

Silvestrini, Lucia; Rossi, Beatrice; Gallmetzer, Andreas; Mathieu, Martine; Scazzocchio, Claudio; Berardi, Enrico; Strauss, Joseph

2015-01-01

A few yeasts, including Hansenula polymorpha are able to assimilate nitrate and use it as nitrogen source. The genes necessary for nitrate assimilation are organised in this organism as a cluster comprising those encoding nitrate reductase (YNR1), nitrite reductase (YNI1), a high affinity transporter (YNT1), as well as the two pathway specific Zn(II)2Cys2 transcriptional activators (YNA1, YNA2). Yna1p and Yna2p mediate induction of the system and here we show that their functions are interdependent. Yna1p activates YNA2 as well as its own (YNA1) transcription thus forming a nitrate-dependent autoactivation loop. Using a split-YFP approach we demonstrate here that Yna1p and Yna2p form a heterodimer independently of the inducer and despite both Yna1p and Yna2p can occupy the target promoter as mono- or homodimer individually, these proteins are transcriptionally incompetent. Subsequently, the transcription factors target genes containing a conserved DNA motif (termed nitrate-UAS) determined in this work by in vitro and in vivo protein-DNA interaction studies. These events lead to a rearrangement of the chromatin landscape on the target promoters and are associated with the onset of transcription of these target genes. In contrast to other fungi and plants, in which nuclear accumulation of the pathway-specific transcription factors only occur in the presence of nitrate, Yna1p and Yna2p are constitutively nuclear in H. polymorpha. Yna2p is needed for this nuclear accumulation and Yna1p is incapable of strictly positioning in the nucleus without Yna2p. In vivo DNA footprinting and ChIP analyses revealed that the permanently nuclear Yna1p/Yna2p heterodimer only binds to the nitrate-UAS when the inducer is present. The nitrate-dependent up-regulation of one partner protein in the heterodimeric complex is functionally similar to the nitrate-dependent activation of nuclear accumulation in other systems.

Physiological evidence for a sodium-dependent high-affinity phosphate and nitrate transport at the plasma membrane of leaf and root cells of Zostera marina L.

PubMed

Rubio, L; Linares-Rueda, A; García-Sánchez, M J; Fernández, J A

2005-02-01

Zostera marina L. is an angiosperm that grows in a medium in which inorganic phosphate (P(i)) and nitrate (NO(3)(-)) are present in micromolar concentrations and must be absorbed against a steep electrochemical potential gradient. The operation of a Na(+)-dependent NO(3)(-) transport was previously demonstrated in leaf cells of this plant, suggesting that other Na(+)-coupled systems could mediate the uptake of anions. To address this question, P(i) transport was studied in leaves and roots of Z. marina, as well as NO(3)(-) uptake in roots. Electrophysiological studies demonstrated that micromolar concentrations of P(i) induced depolarizations of the plasma membrane of root cells. However, this effect was not observed in leaf cells. P(i)-induced depolarizations showed Michaelis-Menten kinetics (K(m)=1.5+/-0.6 microM P(i); D(max)=7.8+/-0.8 mV), and were not observed in the absence of Na(+). However, depolarizations were restored when Na(+) was resupplied. NO(3)(-) additions also evoked depolarizations of the plasma membrane of root cells only in the presence of Na(+). Both NO(3)(-)- and P(i)-induced depolarizations were accompanied by an increase in cytoplasmic Na(+) activity, detected by Na(+)-sensitive microelectrodes. P(i) net uptake (measured in depletion experiments) was stimulated by Na(+). These results strongly suggest that P(i) uptake in roots of Z. marina is mediated by a high-affinity Na(+)-dependent transport system. Both NO(3)(-) and P(i) transport systems exploit the steep inwardly directed electrochemical potential gradient for Na(+), considering the low cytoplasmic Na(+) activity (10.7+/-3.3 mM Na(+)) and the high external Na(+) concentration (500 mM Na(+)).

Knockdown of the partner protein OsNAR2.1 for high-affinity nitrate transport represses lateral root formation in a nitrate-dependent manner

PubMed Central

Huang, Shuangjie; Chen, Si; Liang, Zhihao; Zhang, Chenming; Yan, Ming; Chen, Jingguang; Xu, Guohua; Fan, Xiaorong; Zhang, Yali

2015-01-01

The morphological plasticity of root systems is critical for plant survival, and understanding the mechanisms underlying root adaptation to nitrogen (N) fluctuation is critical for sustainable agriculture; however, the molecular mechanism of N-dependent root growth in rice remains unclear. This study aimed to identify the role of the complementary high-affinity NO3− transport protein OsNAR2.1 in NO3−-regulated rice root growth. Comparisons with wild-type (WT) plants showed that knockdown of OsNAR2.1 inhibited lateral root (LR) formation under low NO3− concentrations, but not under low NH4+ concentrations. 15N-labelling NO3− supplies (provided at concentrations of 0–10 mM) demonstrated that (i) defects in LR formation in mutants subjected to low external NO3− concentrations resulted from impaired NO3− uptake, and (ii) the mutants had significantly fewer LRs than the WT plants when root N contents were similar between genotypes. LR formation in osnar2.1 mutants was less sensitive to localised NO3− supply than LR formation in WT plants, suggesting that OsNAR2.1 may be involved in a NO3−-signalling pathway that controls LR formation. Knockdown of OsNAR2.1 inhibited LR formation by decreasing auxin transport from shoots to roots. Thus, OsNAR2.1 probably functions in both NO3− uptake and NO3−-signalling. PMID:26644084

High-Affinity Methanotrophy Informed by Genome-Wide Analysis of Upland Soil Cluster Alpha (USCα) from Axel Heiberg Island, Canadian High Arctic

NASA Astrophysics Data System (ADS)

Rusley, C.; Onstott, T. C.; Lau, M.

2017-12-01

Methane (CH4) is a potent greenhouse gas whose proper budgeting is vital to climate predictions. Recent studies have identified upland Arctic mineral cryosols as consistent CH4 sinks, drawing CH4 from both the atmosphere and underlying anaerobic soil layers. Global atmospheric CH4 uptake is proposed to be mediated by high-affinity methanotrophs based on the detection of the marker gene pmoA (particulate methane monooxygenase beta subunit). However, a lack of pure cultures and scarcity of genomic information have hindered our understanding of their metabolic capabilities and versatility. Together with the missing genetic linkage between its pmoA and 16S ribosomal RNA (rRNA) gene, the factors that control the distribution and magnitude of high-affinity methanotrophy in the Arctic permafrost-affected region have remained elusive. Using 21 metagenomic datasets of surface soils obtained from long-term core incubation experiments,1 this bioinformatics study aimed to reconstruct the draft genome of the Upland Soil Cluster α-proteobacteria (USCα), the high-affinity methanotroph previously detected in the samples,2 and to determine its phylogeny and metabolic requirements. We obtained a genome bin containing the high-affinity form of the USCα-like pmoA gene. The 3.03 Mbp assembly is 91.6% complete with a unique set of single-copy marker genes. The 16S rRNA gene fragment of USCα belongs to the α-proteobacterial family Beijerinckiaceae. Genome annotation indicates possible formaldehyde oxidation via tetrahydromethanopterin-linked C1 transfer pathways, acetate utilization, carbon fixation via the Calvin-Benson-Bassham cycle, and glycogen production. Notably, the key enzymes for formaldehyde assimilation via the serine and ribulose monophosphate pathways are missing. The presence of genes encoding nitrate reductase and hemoglobin suggests adaptation to low O2 under water-logged conditions. Since USCα has versatile carbon metabolisms, it may not be an obligate methanotroph. Stackhouse, B. T. et al. J. Geophys. Res. Biogeosciences 120, 1764-1784 (2015). Lau, M. C. Y. et al. ISME J. 9, 1880-1891 (2015).

Nitrate assimilation pathway (NAP): role of structural (nit) and transporter (ntr1) genes in Fusarium oxysporum f.sp. lycopersici growth and pathogenicity.

PubMed

Gomez-Gil, Lucia; Camara Almiron, Jesus; Rodriguez Carrillo, Patricia Lizett; Olivares Medina, Cindy Nayely; Bravo Ruiz, Gustavo; Romo Rodriguez, Pamela; Corrales Escobosa, Alma Rosa; Gutierrez Corona, Felix; Roncero, M Isabel

2018-04-01

The tomato pathogen Fusarium oxysporum f.sp. lycopersici possesses the capability to use nitrate as the only nitrogen source under aerobic and anaerobic conditions and to activate virulence-related functions when cultivated in the presence of nitrate, but not in ammonium. The genome of F. oxysporum f.sp. lycopersici encodes three paralogs nitrate reductase (NR) genes (nit1, nit2 and nit3) and one predicted ortholog of the Aspergillus nidulans high-affinity nitrate/nitrite transporters NtrA and NtrB, named ntr1. We set out to clarify the role of nit1, nit2, nit3 and ntr1 genes in nitrate assimilation and in the virulence of F. oxysporum f.sp. lycopersici. Quantitative RT-PCR analysis revealed that only nit1, nit2 and ntr1 are expressed at significant levels during growth in nitrate as the only nitrogen source. Targeted deletion of nit1 and ntr1, but not of nit2 or nit3, severely impaired growth of F. oxysporum on nitrate as nitrogen source, indicating that Nit1 and Ntr1 proteins are involved in nitrate assimilation by the fungus; biochemical analysis of nit mutants indicated that Nit1 and Nit2 enzymes contribute to about 50 and 30% of the total NR activity, respectively. In addition, a spontaneous chlorate-resistant mutant derived from F. oxysporum 4287, denoted NitFG, was characterized, showing inability to grow in nitrate under aerobic and anaerobic conditions and low levels of NR activity, in spite of its increased transcription levels of nit1 and nit2 genes. Tomato plant infection assays showed that NitFG and ∆ntr1 mutants induced an earlier death in tomato plants, whereas the single mutants ∆nit1, ∆nit2 and ∆nit1∆nit2 double mutant showed a mortality rate similar to the wild-type strain. Taken together, these results indicate that the Nit1 and Ntr1 proteins are important for nitrate assimilation of F. oxysporum f.sp. lycopersici incubated under aerobic and anaerobic conditions and that this metabolic process is not essential for the virulence of the fungus. These observations open new questions about the role of Nit1, Nit2, and Nit3 proteins in other routes of nitrate metabolism in this pathogenic fungus and in the possible regulatory role that can be exerted by the AreA protein in these routes.

Elucidating the Role of Site-Specific Nitration of α-Synuclein in the Pathogenesis of Parkinson's Disease via Protein Semisynthesis and Mutagenesis.

PubMed

Burai, Ritwik; Ait-Bouziad, Nadine; Chiki, Anass; Lashuel, Hilal A

2015-04-22

Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra and the presence of intraneuronal inclusions consisting of aggregated and post-translationally modified α-synuclein (α-syn). Despite advances in the chemical synthesis of α-syn and other proteins, the generation of site-specifically nitrated synthetic proteins has not been reported. Consequently, it has not been possible to determine the roles of nitration at specific residues in regulating the physiological and pathogenic properties of α-syn. Here we report, for the first time, the site-specific incorporation of 3-nitrotyrosine at different regions of α-syn using native chemical ligation combined with a novel desulfurization strategy. This strategy enabled us to investigate the role of nitration at single or multiple tyrosine residues in regulating α-syn structure, membrane binding, oligomerization, and fibrils formation. We demonstrate that different site-specifically nitrated α-syn species exhibit distinct structural and aggregation properties and exhibit reduced affinity to negatively charged vesicle membranes. We provide evidence that intermolecular interactions between the N- and C-terminal regions of α-syn play critical roles in mediating nitration-induced α-syn oligomerization. For example, when Y39 is not available for nitration (Y39F and Y39/125F), the extent of cross-linking is limited mostly to dimer formation, whereas mutants in which Y39 along with one or multiple C-terminal tyrosines (Y125F, Y133F, Y136F and Y133/136F) can still undergo nitration readily to form higher-order oligomers. Our semisynthetic strategy for generating site-specifically nitrated proteins opens up new possibilities for investigating the role of nitration in regulating protein structure and function in health and disease.

Nitrification and Autotrophic Nitrifying Bacteria in a Hydrocarbon-Polluted Soil

PubMed Central

Deni, Jamal; Penninckx, Michel J.

1999-01-01

In vitro ammonia-oxidizing bacteria are capable of oxidizing hydrocarbons incompletely. This transformation is accompanied by competitive inhibition of ammonia monooxygenase, the first key enzyme in nitrification. The effect of hydrocarbon pollution on soil nitrification was examined in situ. In a microcosm study, adding diesel fuel hydrocarbon to an uncontaminated soil (agricultural unfertilized soil) treated with ammonium sulfate dramatically reduced the amount of KCl-extractable nitrate but stimulated ammonium consumption. In a soil with long history of pollution that was treated with ammonium sulfate, 90% of the ammonium was transformed into nitrate after 3 weeks of incubation. Nitrate production was twofold higher in the contaminated soil than in the agricultural soil to which hydrocarbon was not added. To assess if ammonia-oxidizing bacteria acquired resistance to inhibition by hydrocarbon, the contaminated soil was reexposed to diesel fuel. Ammonium consumption was not affected, but nitrate production was 30% lower than nitrate production in the absence of hydrocarbon. The apparent reduction in nitrification resulted from immobilization of ammonium by hydrocarbon-stimulated microbial activity. These results indicated that the hydrocarbon inhibited nitrification in the noncontaminated soil (agricultural soil) and that ammonia-oxidizing bacteria in the polluted soil acquired resistance to inhibition by the hydrocarbon, possibly by increasing the affinity of nitrifying bacteria for ammonium in the soil. PMID:10473409

Airway Peroxidases Catalyze Nitration of the β2-Agonist Salbutamol and Decrease Its Pharmacological Activity

PubMed Central

Sallans, Larry; Macha, Stephen; Brown, Kari; McGraw, Dennis W.; Kovacic, Melinda Butsch; Britigan, Bradley E.

2011-01-01

β2-Agonists are the most effective bronchodilators for the rapid relief of asthma symptoms, but for unclear reasons, their effectiveness may be decreased during severe exacerbations. Because peroxidase activity and nitrogen oxides are increased in the asthmatic airway, we examined whether salbutamol, a clinically important β2-agonist, is subject to potentially inactivating nitration. When salbutamol was exposed to myeloperoxidase, eosinophil peroxidase or lactoperoxidase in the presence of hydrogen peroxide (H2O2) and nitrite (NO2−), both absorption spectroscopy and mass spectrometry indicated formation of a new metabolite with features expected for the nitrated drug. The new metabolites showed an absorption maximum at 410 nm and pKa of 6.6 of the phenolic hydroxyl group. In addition to nitrosalbutamol (m/z 285.14), a salbutamol-derived nitrophenol, formed by elimination of the formaldehyde group, was detected (m/z 255.13) by mass spectrometry. It is noteworthy that the latter metabolite was detected in exhaled breath condensates of asthma patients receiving salbutamol but not in unexposed control subjects, indicating the potential for β2-agonist nitration to occur in the inflamed airway in vivo. Salbutamol nitration was inhibited in vitro by ascorbate, thiocyanate, and the pharmacological agents methimazole and dapsone. The efficacy of inhibition depended on the nitrating system, with the lactoperoxidase/H2O2/NO2− being the most affected. Functionally, nitrated salbutamol showed decreased affinity for β2-adrenergic receptors and impaired cAMP synthesis in airway smooth muscle cells compared with the native drug. These results suggest that under inflammatory conditions associated with asthma, phenolic β2-agonists may be subject to peroxidase-catalyzed nitration that could potentially diminish their therapeutic efficacy. PMID:20974700

Impact of iron on silicon utilization by diatoms in the Southern Ocean: A case study of Si/N cycle decoupling in a naturally iron-enriched area

NASA Astrophysics Data System (ADS)

Mosseri, Julie; Quéguiner, Bernard; Armand, Leanne; Cornet-Barthaux, Véronique

2008-03-01

Biogenic silica stocks and fluxes were investigated in austral summer over the naturally iron-fertilized Kerguelen Plateau and in nearby high-nutrient, low-chlorophyll (HNLC) off-plateau surface waters. The Kerguelen Plateau hosted a large-diatom bloom, with high levels of biogenic silica (BSi) but relatively low silicic acid (Si(OH) 4) uptake rates (1100±600 mmol m -2 and 8±4 mmol m -2 d -1, respectively). Diatoms of the naturally iron-enriched area presented high affinities for silicic acid, allowing them in combination with a beneficial nutrient vertical supply to grow in low silicic acid waters (<2 μM). Si(OH) 4 acid uptake rates were also compared with carbon and nitrogen uptake rates. As expected for diatoms growing in favourable nutrient conditions, and from previous artificial iron-enrichment experiments, Si:C and Si:NO 3 elemental uptake ratios of the natural diatom community of the plateau were close to 0.13 and 1, respectively. In contrast, diatom communities in the HNLC waters were composed of strongly silicified (high Si:C, Si:NO 3 uptake ratios) diatoms with low affinities for Si(OH) 4. Although the Si:NO 3 uptake ratio in the surface waters of the plateau was close to 1, the apparent consumption of nitrate on a seasonal basis was much lower (˜5 μM) than the apparent consumption of silicic acid (˜15 μM). This was mainly due to diatoms growing actively on ammonium (i.e. 39-77% of the total nitrogen uptake) produced by an intense heterotrophic activity. Thus we find that while Fe fertilization does increase N uptake with respect to Si uptake, rapid recycling of N decouples nitrogen and carbon export from silica export so that the "silicate pump" remains more efficient than that of N (or P). For this reason an iron-fertilized Southern Ocean is unlikely to experience nitrate exhaustion or export silicic acid to the global ocean.

A new formulation containing calixarene molecules as an emergency treatment of uranium skin contamination.

PubMed

Spagnul, Aurélie; Bouvier-Capely, Céline; Phan, Guillaume; Rebière, François; Fattal, Elias

2010-09-01

Cutaneous contamination represents the second highest contamination pathway in the nuclear industry. Despite that the entry of actinides such as uranium into the body through intact or wounded skin can induce a high internal exposure, no specific emergency treatment for cutaneous contamination exists. In the present work, an innovative formulation dedicated to uranium skin decontamination was developed. The galenic form consists in an oil-in-water nanoemulsion, which contains a tricarboxylic calixarene known for its high uranium affinity and selectivity. The physicochemical characterization of this topical form revealed that calixarene molecules are located at the surface of the dispersed oil droplets of the nanoemulsion, being thus potentially available for uranium chelation. It was demonstrated in preliminary in vitro experiments by using an adapted ultrafiltration method that the calixarene nanoemulsion was able to extract and retain more than 80% of uranium from an aqueous uranyl nitrate contamination solution. First ex vivo experiments carried out in Franz diffusion cells on pig ear skin explants during 24 h showed that the immediate application of the calixarene nanoemulsion on a skin contaminated by a uranyl nitrate solution allowed a uranium transcutaneous diffusion decrease of about 98% through intact and excoriated skins. The calixarene nanoemulsion developed in this study thus seems to be an efficient emergency system for uranium skin decontamination.

Simple guanidinium motif for the selective binding and extraction of sulfate

DOE PAGES

Seipp, Charles A.; Williams, Neil J.; Bryantsev, Vyacheslav S.; ...

2017-06-30

A simple bidentate anion receptor, shown previously to adopt a rigid pseudobicyclic conformation while binding anions in the solid state, selectively binds sulfate in aqueous solutions with logK1 and logK2 values of 3.78 ± 0.12 M-1 and 2.10 ± 0.23 M-1, respectively. This anion receptor has little to no affinity for nitrate and chloride in the same solutions. A lipophilic derivative was synthesized in four steps to yield an extractant that is capable of partitioning sulfate into 1,2 dichloroethane from water in the presence of large excesses of chloride. This extractant demonstrated D values as high as 2.5 with onlymore » 30 mM of anion receptor.« less

Simple guanidinium motif for the selective binding and extraction of sulfate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seipp, Charles A.; Williams, Neil J.; Bryantsev, Vyacheslav S.

A simple bidentate anion receptor, shown previously to adopt a rigid pseudobicyclic conformation while binding anions in the solid state, selectively binds sulfate in aqueous solutions with logK1 and logK2 values of 3.78 ± 0.12 M-1 and 2.10 ± 0.23 M-1, respectively. This anion receptor has little to no affinity for nitrate and chloride in the same solutions. A lipophilic derivative was synthesized in four steps to yield an extractant that is capable of partitioning sulfate into 1,2 dichloroethane from water in the presence of large excesses of chloride. This extractant demonstrated D values as high as 2.5 with onlymore » 30 mM of anion receptor.« less

Plasma nitrate and nitrite are increased by a high nitrate supplement, but not by high nitrate foods in older adults

PubMed Central

Miller, Gary D.; Marsh, Anthony P.; Dove, Robin W.; Beavers, Daniel; Presley, Tennille; Helms, Christine; Bechtold, Erika; King, S. Bruce; Kim-Shapiro, Daniel

2012-01-01

Little is known about the effect of dietary nitrate on the nitrate/nitrite/NO (nitric oxide) cycle in older adults. We examined the effect of a 3-day control diet vs. high nitrate diet, with and without a high nitrate supplement (beetroot juice), on plasma nitrate and nitrite kinetics, and blood pressure using a randomized four period cross-over controlled design. We hypothesized that the high nitrate diet would show higher levels of plasma nitrate/nitrite and blood pressure compared to the control diet, which would be potentiated by the supplement. Participants were eight normotensive older men and women (5 female, 3 male, 72.5±4.7 yrs) with no overt disease or medications that affect NO metabolism. Plasma nitrate and nitrite levels and blood pressure were measured prior to and hourly for 3 hours after each meal. The mean daily changes in plasma nitrate and nitrite were significantly different from baseline for both control diet+supplement (p<0.001 and =0.017 for nitrate and nitrite, respectively) and high nitrate diet+supplement (p=0.001 and 0.002), but not for control diet (p=0.713 and 0.741) or high nitrate diet (p=0.852 and 0.500). Blood pressure decreased from the morning baseline measure to the three 2 hr post-meal follow-up time-points for all treatments, but there was no main effect for treatment. In healthy older adults, a high nitrate supplement consumed at breakfast elevated plasma nitrate and nitrite levels throughout the day. This observation may have practical utility for the timing of intake of a nitrate supplement with physical activity for older adults with vascular dysfunction. PMID:22464802

Short-term effects of a high nitrate diet on nitrate metabolism in healthy individuals.

PubMed

Bondonno, Catherine P; Liu, Alex H; Croft, Kevin D; Ward, Natalie C; Puddey, Ian B; Woodman, Richard J; Hodgson, Jonathan M

2015-03-12

Dietary nitrate, through the enterosalivary nitrate-nitrite-NO pathway, can improve blood pressure and arterial stiffness. How long systemic nitrate and nitrite remain elevated following cessation of high nitrate intake is unknown. In 19 healthy men and women, the time for salivary and plasma nitrate and nitrite to return to baseline after 7 days increased nitrate intake from green leafy vegetables was determined. Salivary and plasma nitrate and nitrite was measured at baseline [D0], end of high nitrate diet [D7], day 9 [+2D], day 14 [+7D] and day 21 [+14D]. Urinary nitrite and nitrate was assessed at D7 and +14D. Increased dietary nitrate for 7 days resulted in a more than fourfold increase in saliva and plasma nitrate and nitrite (p < 0.001) measured at [D7]. At [+2D] plasma nitrite and nitrate had returned to baseline while saliva nitrate and nitrite were more than 1.5 times higher than at baseline levels. By [+7D] all metabolites had returned to baseline levels. The pattern of response was similar between men and women. Urinary nitrate and nitrate was sevenfold higher at D7 compared to +14D. These results suggest that daily ingestion of nitrate may be required to maintain the physiological changes associated with high nitrate intake.

Vinyl sulfone silica: application of an open preactivated support to the study of transnitrosylation of plant proteins by S-nitrosoglutathione

PubMed Central

2013-01-01

Background S-nitrosylaton is implicated in the regulation of numerous signaling pathways with a diversity of regulatory roles. The high lability of the S-NO bond makes the study of proteins regulated by S-nitrosylation/denitrosylation a challenging task and most studies have focused on already S-nitrosylated proteins. We hypothesize that: i) S-nitrosoglutathione (GSNO) transnitrosylation is a feasible mechanism to account for the physiological S-nitrosylation of rather electropositive sulfur atoms from proteins, ii) affinity chromatography is a suitable approach to isolate proteins that are prone to undergo S-transnitrosylation and iii) vinyl sulfone silica is a suitable chromatographic bead. Results The combination of vinyl sulfone silica with GSNO yielded an affinity resin that withstood high ionic strength without shrinking or deforming and that it was suitable to isolate potential GSNO transnitrosylation target candidates. Fractions eluted at 1500 mM NaCl resulted in a symmetrical peak for both, protein and S-nitrosothiols, supporting the idea of transnitrosylation by GSNO as a selective process that involves strong and specific interactions with the target protein. Proteomic analysis led to the identification of 22 physiological significant enzymes that differ with the tissue analyzed, being regulatory proteins the most abundant group in hypocotyls. The identification of chloroplastidic FBPase, proteasome, GTP-binding protein, heat shock Hsp70, syntaxin, catalase I, thioredoxin peroxidase and cytochrome P450 that have already been reported as S-nitrosylated by other techniques can be considered as internal positive controls that validate our experimental approach. An additional validation was provided by the prediction of the S-nitrosylation sites in 19 of the GSNO transnitrosylation target candidates. Conclusions Vinyl sulfone silica is an open immobilization support that can be turned ad hoc and in a straightforward manner into an affinity resin. Its potential in omic sciences was successfully put to test in the context of the analysis of post-translational modification by S-nitrosylation with two different tissues: mature pea leaves and embryogenic sunflower hypocotyls. The identified proteins reveal an intriguing overlap among S-nitrosylation and both tyrosine nitration and thioredoxin regulation. Chloroplastidic FBPase is a paradigm of such overlap of post-translational modifications since it is reversible modified by thioredoxin and S-nitrosylation and irreversibly by tyrosine nitration. Our results suggest a complex interrelation among different modulation mechanisms mediated by NO-derived molecules. PMID:23586608

Sphagnum Mosses - Masters of Efficient N-Uptake while Avoiding Intoxication

PubMed Central

Fritz, Christian; Lamers, Leon P. M.; Riaz, Muhammad; van den Berg, Leon J. L.; Elzenga, Theo J. T. M.

2014-01-01

Peat forming Sphagnum mosses are able to prevent the dominance of vascular plants under ombrotrophic conditions by efficiently scavenging atmospherically deposited nitrogen (N). N-uptake kinetics of these mosses are therefore expected to play a key role in differential N availability, plant competition, and carbon sequestration in Sphagnum peatlands. The interacting effects of rain N concentration and exposure time on moss N-uptake rates are, however, poorly understood. We investigated the effects of N-concentration (1, 5, 10, 50, 100, 500 µM), N-form (15N - ammonium or nitrate) and exposure time (0.5, 2, 72 h) on uptake kinetics for Sphagnum magellanicum from a pristine bog in Patagonia (Argentina) and from a Dutch bog exposed to decades of N-pollution. Uptake rates for ammonium were higher than for nitrate, and N-binding at adsorption sites was negligible. During the first 0.5 h, N-uptake followed saturation kinetics revealing a high affinity (Km 3.5–6.5 µM). Ammonium was taken up 8 times faster than nitrate, whereas over 72 hours this was only 2 times. Uptake rates decreased drastically with increasing exposure times, which implies that many short-term N-uptake experiments in literature may well have overestimated long-term uptake rates and ecosystem retention. Sphagnum from the polluted site (i.e. long-term N exposure) showed lower uptake rates than mosses from the pristine site, indicating an adaptive response. Sphagnum therefore appears to be highly efficient in using short N pulses (e.g. rainfall in pristine areas). This strategy has important ecological and evolutionary implications: at high N input rates, the risk of N-toxicity seems to be reduced by lower uptake rates of Sphagnum, at the expense of its long-term filter capacity and related competitive advantage over vascular plants. As shown by our conceptual model, interacting effects of N-deposition and climate change (changes in rainfall) will seriously alter the functioning of Sphagnum peatlands. PMID:24416125

Sphagnum mosses--masters of efficient N-uptake while avoiding intoxication.

PubMed

Fritz, Christian; Lamers, Leon P M; Riaz, Muhammad; van den Berg, Leon J L; Elzenga, Theo J T M

2014-01-01

Peat forming Sphagnum mosses are able to prevent the dominance of vascular plants under ombrotrophic conditions by efficiently scavenging atmospherically deposited nitrogen (N). N-uptake kinetics of these mosses are therefore expected to play a key role in differential N availability, plant competition, and carbon sequestration in Sphagnum peatlands. The interacting effects of rain N concentration and exposure time on moss N-uptake rates are, however, poorly understood. We investigated the effects of N-concentration (1, 5, 10, 50, 100, 500 µM), N-form ((15)N-ammonium or nitrate) and exposure time (0.5, 2, 72 h) on uptake kinetics for Sphagnum magellanicum from a pristine bog in Patagonia (Argentina) and from a Dutch bog exposed to decades of N-pollution. Uptake rates for ammonium were higher than for nitrate, and N-binding at adsorption sites was negligible. During the first 0.5 h, N-uptake followed saturation kinetics revealing a high affinity (Km 3.5-6.5 µM). Ammonium was taken up 8 times faster than nitrate, whereas over 72 hours this was only 2 times. Uptake rates decreased drastically with increasing exposure times, which implies that many short-term N-uptake experiments in literature may well have overestimated long-term uptake rates and ecosystem retention. Sphagnum from the polluted site (i.e. long-term N exposure) showed lower uptake rates than mosses from the pristine site, indicating an adaptive response. Sphagnum therefore appears to be highly efficient in using short N pulses (e.g. rainfall in pristine areas). This strategy has important ecological and evolutionary implications: at high N input rates, the risk of N-toxicity seems to be reduced by lower uptake rates of Sphagnum, at the expense of its long-term filter capacity and related competitive advantage over vascular plants. As shown by our conceptual model, interacting effects of N-deposition and climate change (changes in rainfall) will seriously alter the functioning of Sphagnum peatlands.

Adsorptive Removal of Nitrate from Aqueous Solution Using Nitrogen Doped Activated Carbon.

PubMed

Machida, Motoi; Goto, Tatsuru; Amano, Yoshimasa; Iida, Tatsuya

2016-01-01

Activated carbon (AC) has been widely applied for adsorptive removal of organic contaminants from aqueous phase, but not for ionic pollutants. In this study, nitrogen doped AC was prepared to increase the adsorption capacity of nitrate from water. AC was oxidized with (NH 4 ) 2 S 2 O 8 solution to maximize oxygen content for the first step, and then NH 3 gas treatment was carried out at 950°C to aim at forming quaternary nitrogen (N-Q) species on AC surface (Ox-9.5AG). Influence of solution pH was examined so as to elucidate the relationship between surface charge and adsorption amounts of nitrate. The results showed that Ox-9.5AG exhibited about twice higher adsorption capacity than non-treatment AC at any initial nitrate concentration and any equilibrium solution pH (pH e ) investigated. The more decrease in pH e value, the more adsorption amount of negatively charged nitrate ion, because the surface charge of AC and Ox-9.5AG could become more positive in acidic solution. The oxidation and consecutive ammonia treatments lead to increase in nitrogen content from 0.35 to 6.4% and decrease in the pH of the point of zero charge (pH pzc ) from 7.1 to 4.0 implying that positively charged N-Q of a Lewis acid was created on the surface of Ox-9.5AG. Based on a Langmuir data analysis, maximum adsorption capacity attained 0.5-0.6 mmol/g of nitrate and adsorption affinity was 3.5-4.0 L/mmol at pH e 2.5 for Ox-9.5AG.

Spatiotemporal variation of nitrate uptake kinetics within the maize (Zea mays L.) root system is associated with greater nitrate uptake and interactions with architectural phenes.

PubMed

York, Larry M; Silberbush, Moshe; Lynch, Jonathan P

2016-06-01

Increasing maize nitrogen acquisition efficiency is a major goal for the 21st century. Nitrate uptake kinetics (NUK) are defined by I max and K m, which denote the maximum uptake rate and the affinity of transporters, respectively. Because NUK have been studied predominantly at the molecular and whole-root system levels, little is known about the functional importance of NUK variation within root systems. A novel method was created to measure NUK of root segments that demonstrated variation in NUK among root classes (seminal, lateral, crown, and brace). I max varied among root class, plant age, and nitrate deprivation combinations, but was most affected by plant age, which increased I max, and nitrate deprivation time, which decreased I max K m was greatest for crown roots. The functional-structural simulation SimRoot was used for sensitivity analysis of plant growth to root segment I max and K m, as well as to test interactions of I max with root system architectural phenes. Simulated plant growth was more sensitive to I max than K m, and reached an asymptote near the maximum I max observed in the empirical studies. Increasing the I max of lateral roots had the largest effect on shoot growth. Additive effects of I max and architectural phenes on nitrate uptake were observed. Empirically, only lateral root tips aged 20 d operated at the maximum I max, and simulations demonstrated that increasing all seminal and lateral classes to this maximum rate could increase plant growth by as much as 26%. Therefore, optimizing I max for all maize root classes merits attention as a promising breeding goal. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Short-Term Effects of a High Nitrate Diet on Nitrate Metabolism in Healthy Individuals

PubMed Central

Bondonno, Catherine P.; Liu, Alex H.; Croft, Kevin D.; Ward, Natalie C.; Puddey, Ian B.; Woodman, Richard J.; Hodgson, Jonathan M.

2015-01-01

Dietary nitrate, through the enterosalivary nitrate-nitrite-NO pathway, can improve blood pressure and arterial stiffness. How long systemic nitrate and nitrite remain elevated following cessation of high nitrate intake is unknown. In 19 healthy men and women, the time for salivary and plasma nitrate and nitrite to return to baseline after 7 days increased nitrate intake from green leafy vegetables was determined. Salivary and plasma nitrate and nitrite was measured at baseline [D0], end of high nitrate diet [D7], day 9 [+2D], day 14 [+7D] and day 21 [+14D]. Urinary nitrite and nitrate was assessed at D7 and +14D. Increased dietary nitrate for 7 days resulted in a more than fourfold increase in saliva and plasma nitrate and nitrite (p < 0.001) measured at [D7]. At [+2D] plasma nitrite and nitrate had returned to baseline while saliva nitrate and nitrite were more than 1.5 times higher than at baseline levels. By [+7D] all metabolites had returned to baseline levels. The pattern of response was similar between men and women. Urinary nitrate and nitrate was sevenfold higher at D7 compared to +14D. These results suggest that daily ingestion of nitrate may be required to maintain the physiological changes associated with high nitrate intake. PMID:25774606

Npr1 Ser/Thr Protein Kinase Links Nitrogen Source Quality and Carbon Availability with the Yeast Nitrate Transporter (Ynt1) Levels*

PubMed Central

Martín, Yusé; González, Yelvis V.; Cabrera, Elisa; Rodríguez, Celia; Siverio, José M.

2011-01-01

Ynt1, the single high affinity nitrate and nitrite transporter of the yeast Hansenula polymorpha, is regulated by the quality of nitrogen sources. Preferred nitrogen sources cause Ynt1 dephosphorylation, ubiquitinylation, endocytosis, and vacuolar degradation. In contrast, under nitrogen limitation Ynt1 is phosphorylated and sorted to the plasma membrane. We show here the involvement of the Ser/Thr kinase HpNpr1 in Ynt1 phosphorylation and regulation of Ynt1 levels in response to nitrogen source quality and the availability of carbon. In Δnpr1, Ynt1 phosphorylation does not take place, although Ynt1 ubiquitin conjugates increase. As a result, in this strain Ynt1 is sorted to the vacuole, from both plasma membrane and the later biosynthetic pathway in nitrogen-free conditions and nitrate. In contrast, overexpression of NPR1 blocks down-regulation of Ynt1, increasing Ynt1 phosphorylation at Ser-244 and -246 and reducing ubiquitinylation. Furthermore, Npr1 is phosphorylated in response to the preferred nitrogen sources, and indeed it is dephosphorylated in nitrogen-free medium. Under conditions where Npr1 is phosphorylated, Ynt1 is not and vice versa. We show for the first time that carbon starvation leads to Npr1 phosphorylation, whereas Ynt1 is dephosphorylated and degraded in the vacuole. Rapamycin prevents this, indicating a possible role of the target of rapamycin signaling pathway in this process. We concluded that Npr1 plays a key role in adapting Ynt1 levels to the nitrogen quality and availability of a source of carbon. PMID:21652715

Plant Nitrogen Acquisition Under Low Availability: Regulation of Uptake and Root Architecture

PubMed Central

Kiba, Takatoshi; Krapp, Anne

2016-01-01

Nitrogen availability is a major factor determining plant growth and productivity. Plants acquire nitrogen nutrients from the soil through their roots mostly in the form of ammonium and nitrate. Since these nutrients are scarce in natural soils, plants have evolved adaptive responses to cope with the environment. One of the most important responses is the regulation of nitrogen acquisition efficiency. This review provides an update on the molecular determinants of two major drivers of the nitrogen acquisition efficiency: (i) uptake activity (e.g. high-affinity nitrogen transporters) and (ii) root architecture (e.g. low-nitrogen-availability-specific regulators of primary and lateral root growth). Major emphasis is laid on the regulation of these determinants by nitrogen supply at the transcriptional and post-transcriptional levels, which enables plants to optimize nitrogen acquisition efficiency under low nitrogen availability. PMID:27025887

Bis-lactam-1,10-phenanthroline (BLPhen), a New Type of Preorganized Mixed N,O-Donor Ligand That Separates Am(III) over Eu(III) with Exceptionally High Efficiency

DOE PAGES

Jansone-Popova, Santa; Ivanov, Alexander S.; Bryantsev, Vyacheslav S.; ...

2017-05-04

Here, we report a new family of preorganized bislactam- 1,10-phenanthroline (BLPhen) complexants that possess both hard and soft donor atoms within a convergent cavity and show unprecedented extraction strength for the trivalent fblock metal ions. BLPhen ligands with saturated and unsaturated δ-lactam rings have notable differences in their affinity and selectivity for Am(III) over Eu(III), with the latter being the most selective mixed N,O-donor extractant of Am(III) reported to date. Saturated BLPhen was crystallized with five Ln(III) nitrates to form charge-neutral 1:1 complexes in the solid state. DFT calculations further elaborate on the variety of effects that dictate the performancemore » of these preorganized compounds.« less

A comparison of the effect of lead nitrate on rat liver chromatin, DNA and histone proteins in solution.

PubMed

Rabbani-Chadegani, Azra; Abdosamadi, Sayeh; Fani, Nesa; Mohammadian, Shayesteh

2009-06-01

Although lead is widely recognized as a toxic substance in the environment and directly damage DNA, no studies are available on lead interaction with chromatin and histone proteins. In this work, we have examined the effect of lead nitrate on EDTA-soluble chromatin (SE chromatin), DNA and histones in solution using absorption and fluorescence spectroscopy, thermal denaturation and gel electrophoresis techniques. The results demonstrate that lead nitrate binds with higher affinity to chromatin than to DNA and produces an insoluble complex as monitored at 400 nm. Binding of lead to DNA decreases its Tm, increases its fluorescence intensity and exhibits hypochromicity at 210 nm which reveal that both DNA bases and the backbone participate in the lead-DNA interaction. Lead also binds strongly to histone proteins in the absence of DNA. The results suggest that although lead destabilizes DNA structure, in the chromatin, the binding of lead introduces some sort of compaction and aggregation, and the histone proteins play a key role in this aspect. This chromatin condensation, upon lead exposure, in turn may decrease fidelity of DNA, and inhibits DNA and RNA synthesis, the process that introduces lead toxicity at the chromatin level.

Availability of phosphate for phytoplankton and bacteria and of labile organic carbon for bacteria at different pCO2 levels in a mesocosm study

NASA Astrophysics Data System (ADS)

Tanaka, T.; Thingstad, T. F.; Løvdal, T.; Grossart, H.-P.; Larsen, A.; Schulz, K. G.; Riebesell, U.

2007-11-01

Availability of phosphate for phytoplankton and bacteria and of labile organic carbon for bacteria at different pCO2 levels were studied in a mesocosm experiment (PeECE III). Using nutrient-depleted SW Norwegian fjord waters, three different levels of pCO2 (350 μatm: 1×CO2; 750 μatm: 2×CO2; 1050 μatm: 3×CO2) were set up, and nitrate and phosphate were added at the start of the experiment in order to induce a phytoplankton bloom. Despite similar responses of total particulate P concentration and phosphate turnover time at the three different pCO2 levels, the size distribution of particulate P and 33PO4 uptake suggested that phosphate transferred to the >10 μm fraction was greater in the 3×CO2 mesocosm during the first 6-10 days when phosphate concentration was high. During the period of phosphate depletion (after Day 12), specific phosphate affinity and specific alkaline phosphatase activity (APA) suggested a P-deficiency (i.e. suboptimal phosphate supply) but not a P-limitation for the phytoplankton and bacterial community at the three different pCO2 levels. Although specific phosphate affinity and specific APA tended to be higher in 3×CO2 than in 2×CO2 and 1×CO2 mesocosms during the phosphate depletion period, no statistical differences were found. Responses of specific glucose affinity for bacteria were similar at the three different pCO2 levels. Measured specific glucose affinities were consistently much lower than the theoretical maximum predicted from the diffusion-limited model, suggesting that bacterial growth was not limited by the availability of labile dissolved organic carbon. These results suggest that availability of phosphate and glucose was similar at the three different pCO2 levels.

Allostery in a disordered protein: Oxidative modifications to α-Synuclein act distally to regulate membrane binding

PubMed Central

Sevcsik, Eva; Trexler, Adam J.; Dunn, Joanna M.; Rhoades, Elizabeth

2011-01-01

Both oxidative stress and aggregation of the protein α-synuclein (aS) have been implicated as key factors in the etiology of Parkinson’s disease. Specifically, oxidative modifications to aS disrupt its binding to lipid membranes, an interaction considered critical to its native function. Here we seek to provide a mechanistic explanation for this phenomenon by investigating the effects of oxidative nitration of tyrosine residues on the structure of aS and its interaction with lipid membranes. Membrane binding is mediated by the first ~95 residues of aS. We find that nitration of the single tyrosine (Y39) in this domain disrupts binding due to electrostatic repulsion. Moreover, we observe that nitration of the three tyrosines (Y125/133/136) in the C-terminal domain is equally effective in perturbing binding, an intriguing result given that the C-terminus is not thought to interact directly with membranes. Our investigations show that tyrosine nitration results in a change of the conformational states populated by aS in solution, with the most prominent changes occurring in the C-terminal region. These results lead us to suggest that nitration of Y125/133/136 reduces the membrane binding affinity of aS through allosteric coupling by altering the ensemble of conformational states and depopulating those capable of membrane binding. While allostery is a well-established concept for structured proteins, it has only recently been discussed in the context of disordered proteins. We propose that allosteric regulation through modification of specific residues in, or ligand binding to, the C-terminus may even be a general mechanism for modulating aS function. PMID:21491910

MpAMT1;2 from Marchantia polymorpha is a High-Affinity, Plasma Membrane Ammonium Transporter.

PubMed

Guo, Hanqing; Wang, Nu; McDonald, Tami R; Reinders, Anke; Ward, John M

2018-05-01

Plant ammonium transporters in the AMT/MEP/Rh (ammonium transporter/methylammonium and ammonium permease/Rhesus factor) superfamily have only been previously characterized in flowering plants (angiosperms). Plant AMT1s are electrogenic, while plant AMT2s are electroneutral, and MEP and Rh transporters in other organisms are electroneutral. We analyzed the transport function of MpAMT1;2 from the basal land plant Marchantia polymorpha, a liverwort. MpAMT1;2 was shown to localize to the plasma membrane in Marchantia gametophyte thallus by stable transformation using a C-terminal citrine fusion. MpAMT1;2 expression was studied using quantitative real-time PCR and shown to be higher when plants were N deficient and lower when plants were grown on media containing ammonium, nitrate or the amino acid glutamine. Expression in Xenopus oocytes and analysis by electrophysiology revealed that MpAMT1;2 is an electrogenic ammonium transporter with a very high affinity for ammonium (7 µM at pH 5.6 and a membrane potential of -137 mV). A conserved inhibitory phosphorylation site identified in angiosperm AMT1s is also present in all AMT1s in Marchantia. Here we show that a phosphomimetic mutation T475D in MpAMT1;2 completely inhibits ammonium transport activity. The results indicate that MpAMT1;2 may be important for ammonium uptake into cells in the Marchantia thallus.

High temperature two component explosive

DOEpatents

Mars, James E.; Poole, Donald R.; Schmidt, Eckart W.; Wang, Charles

1981-01-01

A two component, high temperature, thermally stable explosive composition comprises a liquid or low melting oxidizer and a liquid or low melting organic fuel. The oxidizer and fuel in admixture are incapable of substantial spontaneous exothermic reaction at temperatures on the order of 475.degree. K. At temperatures on the order of 475.degree. K., the oxidizer and fuel in admixture have an activation energy of at least about 40 kcal/mol. As a result of the high activation energy, the preferred explosive compositions are nondetonable as solids at ambient temperature, and become detonable only when heated beyond the melting point. Preferable oxidizers are selected from alkali or alkaline earth metal nitrates, nitrites, perchlorates, and/or mixtures thereof. Preferred fuels are organic compounds having polar hydrophilic groups. The most preferred fuels are guanidinium nitrate, acetamide and mixtures of the two. Most preferred oxidizers are eutectic mixtures of lithium nitrate, potassium nitrate and sodium nitrate, of sodium nitrite, sodium nitrate and potassium nitrate, and of potassium nitrate, calcium nitrate and sodium nitrate.

Effects of heat shock, stannous chloride, and gallium nitrate on the rat inflammatory response.

PubMed

House, S D; Guidon, P T; Perdrizet, G A; Rewinski, M; Kyriakos, R; Bockman, R S; Mistry, T; Gallagher, R A; Hightower, L E

2001-04-01

Heat and a variety of other stressors cause mammalian cells and tissues to acquire cytoprotection. This transient state of altered cellular physiology is nonproliferative and antiapoptotic. In this study, male Wistar rats were stress conditioned with either stannous chloride or gallium nitrate, which have immunosuppressive effects in vivo and in vitro, or heat shock, the most intensively studied inducer of cytoprotection. The early stages of inflammation in response to topical suffusion of mesentery tissue with formyl-methionyl-leucyl-phenylalanine (FMLP) were monitored using intravital microscopy. Microvascular hemodynamics (venular diameter, red blood cell velocity [Vrbc], white blood cell [WBC] flux, and leukocyte-endothelial adhesion [LEA]) were used as indicators of inflammation, and tissue levels of inducible Hsp70, determined using immunoblot assays, provided a marker of cytoprotection. None of the experimental treatments blocked decreases in WBC flux during FMLP suffusion, an indicator of increased low-affinity interactions between leukocytes and vascular endothelium known as rolling adhesion. During FMLP suffusion LEA, an indicator of firm attachment between leukocytes and vascular endothelial cells increased in placebo and gallium nitrate-treated animals but not in heat- and stannous chloride-treated animals, an anti-inflammatory effect. Hsp70 was not detected in aortic tissue from placebo and gallium nitrate-treated animals, indicating that Hsp70-dependent cytoprotection was not present. In contrast, Hsp70 was detected in aortic tissues from heat- and stannous chloride-treated animals, indicating that these tissues were in a cytoprotected state that was also an anti-inflammatory state.

Determining Roles of Accessory Genes in Denitrification by Mutant Fitness Analyses

PubMed Central

Vaccaro, Brian J.; Thorgersen, Michael P.; Lancaster, W. Andrew; Price, Morgan N.; Wetmore, Kelly M.; Poole, Farris L.; Deutschbauer, Adam; Arkin, Adam P.

2015-01-01

Enzymes of the denitrification pathway play an important role in the global nitrogen cycle, including release of nitrous oxide, an ozone-depleting greenhouse gas. In addition, nitric oxide reductase, maturation factors, and proteins associated with nitric oxide detoxification are used by pathogens to combat nitric oxide release by host immune systems. While the core reductases that catalyze the conversion of nitrate to dinitrogen are well understood at a mechanistic level, there are many peripheral proteins required for denitrification whose basic function is unclear. A bar-coded transposon DNA library from Pseudomonas stutzeri strain RCH2 was grown under denitrifying conditions, using nitrate or nitrite as an electron acceptor, and also under molybdenum limitation conditions, with nitrate as the electron acceptor. Analysis of sequencing results from these growths yielded gene fitness data for 3,307 of the 4,265 protein-encoding genes present in strain RCH2. The insights presented here contribute to our understanding of how peripheral proteins contribute to a fully functioning denitrification pathway. We propose a new low-affinity molybdate transporter, OatABC, and show that differential regulation is observed for two MoaA homologs involved in molybdenum cofactor biosynthesis. We also propose that NnrS may function as a membrane-bound NO sensor. The dominant HemN paralog involved in heme biosynthesis is identified, and a CheR homolog is proposed to function in nitrate chemotaxis. In addition, new insights are provided into nitrite reductase redundancy, nitric oxide reductase maturation, nitrous oxide reductase maturation, and regulation. PMID:26452555

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vaccaro, Brian J.; Thorgersen, Michael P.; Lancaster, W. Andrew

Enzymes of the denitrification pathway play an important role in the global nitrogen cycle, including release of nitrous oxide, an ozone-depleting greenhouse gas. In addition, nitric oxide reductase, maturation factors, and proteins associated with nitric oxide detoxification are used by pathogens to combat nitric oxide release by host immune systems. While the core reductases that catalyze the conversion of nitrate to dinitrogen are well understood at a mechanistic level, there are many peripheral proteins required for denitrification whose basic function is unclear. A bar-coded transposon DNA library fromPseudomonas stutzeristrain RCH2 was grown under denitrifying conditions, using nitrate or nitrite asmore » an electron acceptor, and also under molybdenum limitation conditions, with nitrate as the electron acceptor. Analysis of sequencing results from these growths yielded gene fitness data for 3,307 of the 4,265 protein-encoding genes present in strain RCH2. The insights presented here contribute to our understanding of how peripheral proteins contribute to a fully functioning denitrification pathway. We propose a new low-affinity molybdate transporter, OatABC, and show that differential regulation is observed for two MoaA homologs involved in molybdenum cofactor biosynthesis. We also propose that NnrS may function as a membrane-bound NO sensor. Finally, the dominant HemN paralog involved in heme biosynthesis is identified, and a CheR homolog is proposed to function in nitrate chemotaxis. In addition, new insights are provided into nitrite reductase redundancy, nitric oxide reductase maturation, nitrous oxide reductase maturation, and regulation.« less

Post-anthesis nitrate uptake is critical to yield and grain protein content in Sorghum bicolor.

PubMed

Worland, Belinda; Robinson, Nicole; Jordan, David; Schmidt, Susanne; Godwin, Ian

2017-09-01

Crops only use ∼50% of applied nitrogen (N) fertilizer creating N losses and pollution. Plants need to efficiently uptake and utilize N to meet growing global food demands. Here we investigate how the supply and timing of nitrate affects N status and yield in Sorghum bicolor (sorghum). Sorghum was grown in pots with either 10mM (High) or 1mM (Low) nitrate supply. Shortly before anthesis the nitrate supply was either maintained, increased 10-fold or eliminated. Leaf sheaths of sorghum grown with High nitrate accumulated nitrate in concentrations >3-times higher than leaves. Removal of nitrate supply pre-anthesis resulted in the rapid reduction of stored nitrate in all organs. Plants receiving a 10-fold increase in nitrate supply pre-anthesis achieved similar grain yield and protein content and 29% larger grains than those maintained on High nitrate, despite receiving 24% less nitrate over the whole growth period. In sorghum, plant available N is important throughout development, particularly anthesis and grain filling, for grain yield and grain protein content. Nitrate accumulation in leaf sheaths presents opportunities for the genetic analysis of mechanisms behind nitrate storage and remobilization in sorghum to improve N use efficiency. Copyright © 2017 Elsevier GmbH. All rights reserved.

Habitual Dietary Nitrate Intake in Highly Trained Athletes.

PubMed

Jonvik, Kristin L; Nyakayiru, Jean; van Dijk, Jan-Willem; Wardenaar, Floris C; van Loon, Luc J C; Verdijk, Lex B

2017-04-01

Although beetroot juice, as a nitrate carrier, is a popular ergogenic supplement among athletes, nitrate is consumed through the regular diet as well. We aimed to assess the habitual dietary nitrate intake and identify the main contributing food sources in a large group of highly trained athletes. Dutch highly trained athletes (226 women and 327 men) completed 2-4 web-based 24-hr dietary recalls and questionnaires within a 2- to 4-week period. The nitrate content of food products and food groups was determined systematically based on values found in regulatory reports and scientific literature. These were then used to calculate each athlete's dietary nitrate intake from the web-based recalls. The median[IQR] habitual nitrate intake was 106[75-170] mg/d (range 19-525 mg/d). Nitrate intake correlated with energy intake (ρ = 0.28, p < .001), and strongly correlated with vegetable intake (ρ = 0.78, p < .001). In accordance, most of the dietary nitrate was consumed through vegetables, potatoes and fruit, accounting for 74% of total nitrate intake, with lettuce and spinach contributing most. When corrected for energy intake, nitrate intake was substantially higher in female vs male athletes (12.8[9.2-20.0] vs 9.4[6.2-13.8] mg/MJ; p < .001). This difference was attributed to the higher vegetable intake in female vs male athletes (150[88-236] vs 114[61-183] g/d; p < .001). In conclusion, median daily intake of dietary nitrate in highly trained athletes was 106 mg, with large interindividual variation. Dietary nitrate intake was strongly associated with the intake of vegetables. Increasing the intake of nitrate-rich vegetables in the diet might serve as an alternative strategy for nitrate supplementation.

Specific enrichment of a targeted nitrotyrosine-containing peptide from complex matrices and relative quantification for liquid chromatography-mass spectrometry analysis.

PubMed

Yang, Yun

2017-02-17

Protein tyrosine nitration is considered an important non-enzymatic post-translational modification. In the tyrosine nitration process, 3-nitrotyrosine is formed and recognized as a biomarker of nitrosative/nitrative stress implicated in inflammatory responses and age-related disorders. In view of the complexity of biological samples and the ultra-low abundance of protein-incorporated nitrotyrosine, selective enrichment of nitrotyrosine-containing peptides prior to chromatographic separation is crucial. Herein, I report a simple yet highly specific and efficient enrichment method for nitrotyrosine-containing peptides. After blocking all primary amines in the sample by acetylation with acetic anhydride, I then further converted all nitrotyrosine residues into aminotyrosine residues by reduction with dithiothreitol and hemin. Therefore, I eliminated the side-product with 80Da adduct, since inevitable considerable amount of which was generated in the widely used reduction mediated by sodium dithionite. Both acetylation and reduction yields were close to 100%, and my one-pot sample derivatization applied no solid phase extraction steps or sample transference to avoid sample loss. To capture and release aminotyrosine-containing peptides, I synthesized an N-hydroxysuccinimide-ester-functionalized stationary phase which had very high affinity towards amino groups and possessed a base-cleavable ester linker to retrieve targeted peptides by hydrolysis. I validated this strategy by highly efficient enrichment of the targeted peptide from complex matrices of trypsin-digested bovine serum albumin (BSA) and human plasma spiked with derivatized nitrotyrosine-containing angiotensin II. My enrichment method successfully removed most untargeted peptides in those samples. By relative quantification with home-made identical and stable-isotope labelled internal standards, I investigated the recoveries of a nitrotyrosine-containing peptide from complex biological matrices during enrichment for the first time. Mean recoveries were 49.8% and 41.1% (n=6) for the enrichment of nitrotyrosine-containing angiotensin II from 1:100 (w/w) BSA digest and from 1:10 000 (w/w) human plasma digest, respectively. My enrichment method demonstrated great potential in future applications to clinical samples and biomarker discovery. Copyright © 2017 Elsevier B.V. All rights reserved.

Aminoethyl nitrate – the novel super nitrate?

PubMed Central

Bauersachs, Johann

2009-01-01

Long-term use of most organic nitrates is limited by development of tolerance, induction of oxidative stress and endothelial dysfunction. In this issue of the BJP, Schuhmacher et al. characterized a novel class of organic nitrates with amino moieties (aminoalkyl nitrates). Aminoethyl nitrate was identified as a novel organic mononitrate with high potency but devoid of induction of mitochondrial oxidative stress. Cross-tolerance to nitroglycerin or the endothelium-dependent agonist acetylcholine after in vivo treatment was not observed. Like all nitrates, aminoethyl nitrate induced vasorelaxation by activation of soluble guanylate cyclase. Thus, in contrast to the prevailing view, high potency in an organic nitrate is not necessarily accompanied by induction of oxidative stress or endothelial dysfunction. This work from Daiber's group is an important step forward in the understanding of nitrate bioactivation, tolerance phenomena and towards the development of better organic nitrates for clinical use. PMID:19732062

Characteristics of ammonium and nitrate uptake by phytoplankton in Lake Kinneret

DOE Office of Scientific and Technical Information (OSTI.GOV)

Berman, T.; Sherr, B.F.; Sherr, E.

1984-03-01

Seasonal and diurnal patterns of NH/sub 4//sup +/ and NO/sub 3//sup -/ uptake were determined for Lake Kinneret phytoplankton. Nanoplankton generally, but not always, had a higher uptake of NH/sub 4//sup +/ and NO/sub 3//sup -/ than did net plankton. Ammonium was always taken up preferentially and the phytoplankton had lower apparent affinity constants (K/sub s/) for this ion than for NO/sub 3//sup -/. However, during the annual dinoflagellate bloom of Peridinium (February-May), when ambient levels of NH/sub 4//sup +/ were low and those of NO/sub 3//sup -/ were high, a considerable portion of the total N flux was derivedmore » from NO/sub 3//sup -/. The finding that NO/sub 3//sup -/ was utilized by the dinoflagellates implies that control of the amounts of this nutrient, which is predominantly supplied from watershed sources, could be important in limiting eutrophication in Lake Kinneret.« less

Plant Nitrogen Acquisition Under Low Availability: Regulation of Uptake and Root Architecture.

PubMed

Kiba, Takatoshi; Krapp, Anne

2016-04-01

Nitrogen availability is a major factor determining plant growth and productivity. Plants acquire nitrogen nutrients from the soil through their roots mostly in the form of ammonium and nitrate. Since these nutrients are scarce in natural soils, plants have evolved adaptive responses to cope with the environment. One of the most important responses is the regulation of nitrogen acquisition efficiency. This review provides an update on the molecular determinants of two major drivers of the nitrogen acquisition efficiency: (i) uptake activity (e.g. high-affinity nitrogen transporters) and (ii) root architecture (e.g. low-nitrogen-availability-specific regulators of primary and lateral root growth). Major emphasis is laid on the regulation of these determinants by nitrogen supply at the transcriptional and post-transcriptional levels, which enables plants to optimize nitrogen acquisition efficiency under low nitrogen availability. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

Nitrate-nitrogen levels in rural drinking water: Is there an association with age-related macular degeneration?

PubMed

Klein, Barbara E K; McElroy, Jane A; Klein, Ronald; Howard, Kerri P; Lee, Kristine E

2013-01-01

We examined the association of nitrate-nitrogen exposure from rural private drinking water and incidence of age-related macular degeneration (AMD). All participants in the Beaver Dam Eye Study (53916 improvement plan code) completed a questionnaire and had an ocular examination including standardized, graded fundus photographs at five examinations. Only information from rural residents in that study are included in this report. Data from an environmental monitoring study with probabilistic-based agro-chemical sampling, including nitrate-nitrogen, of rural private drinking water were available. Incidence of early AMD was associated with elevated nitrate-nitrogen levels in rural private drinking water supply (10.0% for low, 19.2% for medium, and 26.1% for high nitrate-nitrogen level in the right eye). The odds ratios (ORs) were 1.77 (95% confidence interval [CI]: 1.12-2.78) for medium and 2.88 (95% CI: 1.59-5.23) for high nitrate-nitrogen level. Incidence of late AMD was increased for those with medium or high levels of nitrate-nitrogen compared to low levels (2.3% for low and 5.1% for the medium or high nitrate-nitrogen level, for the right eye). The OR for medium or high nitrate-nitrogen groups was 2.80 (95% CI: 1.07-7.31) compared to the low nitrate-nitrogen group.

Nitrate-Nitrogen Levels in Rural Drinking Water: Is There an Association with Age-related Macular Degeneration?

PubMed Central

Klein, Barbara E. K.; McElroy, Jane A.; Klein, Ronald; Howard, Kerri P.; Lee, Kristine E.

2014-01-01

We examined the association of nitrate-nitrogen exposure from rural private drinking water and incidence of age-related macular degeneration (AMD). Participants of the Beaver Dam Eye Study living in rural areas within the 53916 zone improvement plan code but outside the city limits of Beaver Dam, Wisconsin (Beaver Dam Township) completed a questionnaire and ocular examination including standardized, graded fundus photographs at five examinations. Data from an environmental monitoring study with probabilistic-based agro-chemical sampling, including nitrate-nitrogen, of rural private drinking water were available. Incidence of early AMD was associated with elevated nitrate-nitrogen levels in rural private drinking water supply (10.0% for low, 19.2% for medium, and 26.1% for high nitrate-nitrogen level in the right eye). The odds ratios (ORs) were 1.77 (95% confidence interval [CI]: 1.12–2.78) for medium and 2.88 (95% CI: 1.59–5.23) for high nitrate-nitrogen level. Incidence of late AMD was increased for those with medium or high levels of nitrate-nitrogen compared to low levels (2.3% for low and 5.1% for the medium or high nitrate-nitrogen level, for the right eye). The OR for medium or high nitrate-nitrogen groups was 2.80 (95% CI: 1.07–7.31) compared to the low nitrate-nitrogen group. PMID:24007430

Interaction of organic carbon, reduced sulphur and nitrate in anaerobic baffled reactor for fresh leachate treatment.

PubMed

Yin, Zhixuan; Xie, Li; Khanal, Samir Kumar; Zhou, Qi

2016-01-01

Interaction of organic carbon, reduced sulphur and nitrate was examined using anaerobic baffled reactor for fresh leachate treatment by supplementing nitrate and/or sulphide to compartment 3. Nitrate was removed completely throughout the study mostly via denitrification (>80%) without nitrite accumulation. Besides carbon source, various reduced sulphur (e.g. sulphide, elemental sulphur and organic sulphur) could be involved in the nitrate reduction process via sulphur-based autotrophic denitrification when dissolved organic carbon/nitrate ratio decreased below 1.6. High sulphide concentration not only stimulated autotrophic denitrification, but it also inhibited heterotrophic denitrification, resulting in a shift (11-20%) from heterotrophic denitrification to dissimilatory nitrate reduction to ammonia. High-throughput 16S rRNA gene sequencing analysis further confirmed that sulphur-oxidizing nitrate-reducing bacteria were stimulated with increase in the proportion of bacterial population from 18.6% to 27.2% by high sulphide concentration, meanwhile, heterotrophic nitrate-reducing bacteria and fermentative bacteria were inhibited with 25.5% and 66.6% decrease in the bacterial population.

California GAMA Special Study: An isotopic and dissolved gas investigation of nitrate source and transport to a public supply well in California's Central Valley

DOE Office of Scientific and Technical Information (OSTI.GOV)

Singleton, M J; Moran, J E; Esser, B K

2010-04-14

This study investigates nitrate contamination of a deep municipal drinking water production well in Ripon, CA to demonstrate the utility of natural groundwater tracers in constraining the sources and transport of nitrate to deep aquifers in the Central Valley. The goal of the study was to investigate the origin (source) of elevated nitrate and the potential for the deep aquifer to attenuate anthropogenic nitrate. The site is ideal for such an investigation. The production well is screened from 165-325 feet below ground surface and a number of nearby shallow and deep monitoring wells were available for sampling. Furthermore, potential sourcesmore » of nitrate contamination to the well had been identified, including a fertilizer supply plant located approximately 1000 feet to the east and local almond groves. A variety of natural isotopic and dissolved gas tracers including {sup 3}H-{sup 3}He groundwater age and the isotopic composition of nitrate are applied to identify nitrate sources and to characterize nitrate transport. An advanced method for sampling production wells is employed to help identify contaminant contributions from specific screen intervals. Nitrate transport: Groundwater nitrate at this field site is not being actively denitrified. Groundwater parameters indicate oxic conditions, the dissolved gas data shows no evidence for excess nitrogen as the result of denitrification, and nitrate-N and -O isotope compositions do not display patterns typical of denitrification. Contaminant nitrate source: The ambient nitrate concentration in shallow groundwater at the Ripon site ({approx}12 mg/L as nitrate) is typical of shallow groundwaters affected by recharge from agricultural and urban areas. Nitrate concentrations in Ripon City Well 12 (50-58 mg/L as nitrate) are significantly higher than these ambient concentrations, indicating an additional source of anthropogenic nitrate is affecting groundwater in the capture zone of this municipal drinking water well. This study provides two new pieces of evidence that the Ripon Farm Services Plant is the source of elevated nitrate in Ripon City Well 12. (1) Chemical mass balance calculations using nitrate concentration, nitrate isotopic composition, and initial tritium activity all indicate that that the source water for elevated nitrate to Ripon City Well 12 is a very small component of the water produced by City Well 12 and thus must have extremely high nitrate concentration. The high source water nitrate concentration ({approx}1500 mg/L as nitrate) required by these mass balance calculations precludes common sources of nitrate such as irrigated agriculture, dairy wastewater, and septic discharge. Shallow groundwater under the Ripon Farm Services RFS plant does contain extremely high concentrations of nitrate (>1700 mg/L as nitrate). (2) Nitrogen and oxygen isotope compositions of nitrate indicate that the additional anthropogenic nitrate source to Ripon City Well 12 is significantly enriched in {delta}{sup 18}O-NO{sub 3}, an isotopic signature consistent with synthetic nitrate fertilizer, and not with human or animal wastewater discharge (i.e. dairy operations, septic system discharge, or municipal wastewater discharge), or with organic fertilizer. Monitoring wells on and near the RFS plant also have high {delta}{sup 18}O-NO{sub 3}, and the plant has handled and stored synthetic nitrate fertilizer that will have this isotopic signature. The results described here highlight the complexity of attributing nitrate found in long screened, high capacity wells to specific sources. In this case, the presence of a very high concentration source near the well site combined with sampling using multiple isotopic tracer techniques and specialized depth-specific techniques allowed fingerprinting of the source in the mixed-age samples drawn from the production well.« less

Effect of Spinach, a High Dietary Nitrate Source, on Arterial Stiffness and Related Hemodynamic Measures: A Randomized, Controlled Trial in Healthy Adults

PubMed Central

Jovanovski, Elena; Bosco, Laura; Khan, Kashif; Au-Yeung, Fei; Ho, Hoang; Zurbau, Andreea; Jenkins, Alexandra L.

2015-01-01

Diets rich in fruits and vegetables reduce risk of adverse cardiovascular events. However, the constituents responsible for this effect have not been well established. Lately, the attention has been brought to vegetables with high nitrate content with evidence that this might represent a source of vasoprotective nitric oxide. We hypothesized that short-term consumption of spinach, a vegetable having high dietary nitrate content, can affect the arterial waveform indicative of arterial stiffness, as well as central and peripheral blood pressure (BP). Using a placebo-controlled, crossover design, 27 healthy participants were randomly assigned to receive either a high-nitrate (spinach; 845 mg nitrate/day) or low-nitrate soup (asparagus; 0.6 mg nitrate/day) for 7 days with a 1-week washout period. On days 1 and 7, profiles of augmentation index, central, and brachial BP were obtained over 180 min post-consumption in 4 fasted visits. A postprandial reduction in augmentation index was observed at 180 min on high-nitrate compared to low-nitrate intervention (-6.54 ± 9.7% vs. -0.82 ± 8.0%, p = 0.01) on Day 1, and from baseline on Day 7 (-6.93 ± 8.7%, p < 0.001; high vs. low: -2.28 ± 12.5%, p = 0.35), suggesting that the nitrate intervention is not associated with the development of tolerance for at least 7 days of continued supplementation. High vs. low-nitrate intervention also reduced central systolic (-3.39 ± 5.6 mmHg, p = 0.004) and diastolic BP (-2.60 ± 5.8 mmHg, p = 0.028) and brachial systolic BP (-3.48 ± 7.4 mmHg, p = 0.022) at 180 min following 7-day supplementation only. These findings suggest that dietary nitrate from spinach may contribute to beneficial hemodynamic effects of vegetable-rich diets and highlights the potential of developing a targeted dietary approach in the management of elevated BP. PMID:26251834

Contribution of atmospheric nitrate to stream-water nitrate in Japanese coniferous forests revealed by the oxygen isotope ratio of nitrate.

PubMed

Tobari, Y; Koba, K; Fukushima, K; Tokuchi, N; Ohte, N; Tateno, R; Toyoda, S; Yoshioka, T; Yoshida, N

2010-05-15

Evaluation of the openness of the nitrogen (N) cycle in forest ecosystems is important in efforts to improve forest management because the N supply often limits primary production. The use of the oxygen isotope ratio (delta(18)O) of nitrate is a promising approach to determine how effectively atmospheric nitrate can be retained in a forest ecosystem. We investigated the delta(18)O of nitrate in stream water in order to estimate the contribution of atmospheric NO(3) (-) in stream-water NO(3) (-) (f(atm)) from 26 watersheds with different stand ages (1-87 years) in Japan. The stream-water nitrate concentrations were high in young forests whereas, in contrast, old forests discharged low-nitrate stream water. These results implied a low f(atm) and a closed N cycle in older forests. However, the delta(18)O values of nitrate in stream water revealed that f(atm) values were higher in older forests than in younger forests. These results indicated that even in old forests, where the discharged N loss was small, atmospheric nitrate was not retained effectively. The steep slopes of the studied watersheds (>40 degrees ) which hinder the capturing of atmospheric nitrate by plants and microbes might be responsible for the inefficient utilization of atmospheric nitrate. Moreover, the unprocessed fraction of atmospheric nitrate in the stream-water nitrate in the forest (f(unprocessed)) was high in the young forest (78%), although f(unprocessed) was stable and low for other forests (5-13%). This high f(unprocessed) of the young forest indicated that the young forest retained neither atmospheric NO(3) (-) nor soil NO(3) (-) effectively, engendering high stream-water NO(3) (-) concentrations. Copyright (c) 2010 John Wiley & Sons, Ltd.

Assessing the Role of Sewers and Atmospheric Deposition as Nitrate Contamination Sources to Urban Surface Waters using Stable Nitrate Isotopes

NASA Astrophysics Data System (ADS)

Sikora, M. T.; Elliott, E. M.

2009-12-01

Excess nitrate (NO3-) contributes to the overall degraded quality of streams in many urban areas. These systems are often dominated by impervious surfaces and storm sewers that can route atmospherically deposited nitrogen, from both wet and dry deposition, to waterways. Moreover, in densely populated watersheds there is the potential for interaction between urban waterways and sewer systems. The affects of accumulated nitrate in riverine and estuary systems include low dissolved oxygen, loss of species diversity, increased mortality of aquatic species, and general eutrophication of the waterbody. However, the dynamics of nitrate pollution from each source and it’s affect on urban waterways is poorly constrained. The isotopes of nitrogen and oxygen in nitrate have been proven effective in helping to distinguish contamination sources to ground and surface waters. In order to improve our understanding of urban nitrate pollution sources and dynamics, we examined nitrate isotopes (δ15N and δ18O) in base- and stormflow samples collected over a two-year period from a restored urban stream in Pittsburgh, Pennsylvania (USA). Nine Mile Run drains a 1,600 hectare urban watershed characterized by 38% impervious surface cover. Prior work has documented high nitrate export from the watershed (~19 kg NO3- ha-1 yr-1). Potential nitrate sources to the watershed include observed sewer overflows draining directly to the stream, as well as atmospheric deposition (~23 kg NO3- ha-1 yr-1). In this and other urban systems with high percentages of impervious surfaces, there is likely minimal input from nitrate derived from soil or fertilizer. In this presentation, we examine spatial and temporal patterns in nitrate isotopic composition collected at five locations along Nine Mile Run characterized by both sanitary and combined-sewer cross-connections. Preliminary isotopic analysis of low-flow winter streamwater samples suggest nitrate export from Nine Mile Run is primarily influenced by inputs of human waste despite high rates of atmospheric nitrate deposition. Further isotopic analysis of nitrate will examine seasonal variations in nitrate sources; compare nitrate dynamics and sources during low- versus high-flows, and the influence of interannual climatic variability on nitrate export.

Root Type-Specific Reprogramming of Maize Pericycle Transcriptomes by Local High Nitrate Results in Disparate Lateral Root Branching Patterns1[OPEN

PubMed Central

Lithio, Andrew

2016-01-01

The adaptability of root system architecture to unevenly distributed mineral nutrients in soil is a key determinant of plant performance. The molecular mechanisms underlying nitrate dependent plasticity of lateral root branching across the different root types of maize are only poorly understood. In this study, detailed morphological and anatomical analyses together with cell type-specific transcriptome profiling experiments combining laser capture microdissection with RNA-seq were performed to unravel the molecular signatures of lateral root formation in primary, seminal, crown, and brace roots of maize (Zea mays) upon local high nitrate stimulation. The four maize root types displayed divergent branching patterns of lateral roots upon local high nitrate stimulation. In particular, brace roots displayed an exceptional architectural plasticity compared to other root types. Transcriptome profiling revealed root type-specific transcriptomic reprogramming of pericycle cells upon local high nitrate stimulation. The alteration of the transcriptomic landscape of brace root pericycle cells in response to local high nitrate stimulation was most significant. Root type-specific transcriptome diversity in response to local high nitrate highlighted differences in the functional adaptability and systemic shoot nitrogen starvation response during development. Integration of morphological, anatomical, and transcriptomic data resulted in a framework underscoring similarity and diversity among root types grown in heterogeneous nitrate environments. PMID:26811190

Low dose dietary nitrate improves endothelial dysfunction and plaque stability in the ApoE-/- mouse fed a high fat diet.

PubMed

Bakker, J R; Bondonno, N P; Gaspari, T A; Kemp-Harper, B K; McCashney, A J; Hodgson, J M; Croft, K D; Ward, N C

2016-10-01

Nitric oxide (NO) is an important vascular signalling molecule. NO is synthesised endogenously by endothelial nitric oxide synthase (eNOS). An alternate pathway is exogenous dietary nitrate, which can be converted to nitrite and then stored or further converted to NO and used immediately. Atherosclerosis is associated with endothelial dysfunction and subsequent lesion formation. This is thought to arise due to a reduction in the bioavailability and/or bioactivity of endogenous NO. To determine if dietary nitrate can protect against endothelial dysfunction and lesion formation in the ApoE -/- mouse fed a high fat diet (HFD). ApoE -/- fed a HFD were randomized to receive (i) high nitrate (10mmol/kg/day, n=12), (ii) moderate nitrate (1mmol/kg/day, n=8), (iii) low nitrate (0.1mmol/kg/day, n=8), or (iv) sodium chloride supplemented drinking water (control, n=10) for 10 weeks. A group of C57BL6 mice (n=6) received regular water and served as a healthy reference group. At 10 weeks, ACh-induced vessel relaxation was significantly impaired in ApoE -/- mice versus C57BL6. Mice supplemented with low or moderate nitrate showed significant improvements in ACh-induced vessel relaxation compared to ApoE -/- mice given the high nitrate or sodium chloride. Plaque collagen expression was increased and lipid deposition reduced following supplementation with low or moderate nitrate compared to sodium chloride, reflecting increased plaque stability with nitrate supplementation. Plasma nitrate and nitrite levels were significantly increased in all three groups fed the nitrate-supplemented water. Low and moderate dose nitrate significantly improved endothelial function and atherosclerotic plaque composition in ApoE -/- mice fed a HFD. Copyright © 2016 Elsevier Inc. All rights reserved.

Formation, Evaporation, and Hydrolysis of Organic Nitrates from Nitrate Radical Oxidation of Monoterpenes

NASA Astrophysics Data System (ADS)

Ng, N. L.; Takeuchi, M.; Eris, G.; Berkemeier, T.; Boyd, C.; Nah, T.; Xu, L.

2017-12-01

Organic nitrates play an important role in the cycling of NOx and secondary organic aerosol (SOA) formation, yet their formation mechanisms and fates remain highly uncertain. The interactions of biogenic VOCs with NO3 radicals represent a direct way for positively linking anthropogenic and biogenic emissions. Results from ambient studies suggest that organic nitrates have a relatively short lifetime, though corresponding laboratory data are limited. SOA and organic nitrates produced at night may evaporate the following morning due to increasing temperatures or dilution of semi-volatile compounds. Once formed, organic nitrates can also undergo hydrolysis in the presence of particle water. In this work, we investigate the formation, evaporation, and hydrolysis of organic nitrates generated from the nitrate radical oxidation of a-pinene, b-pinene, and limonene. Experiments are conducted in the Georgia Environmental Chamber facility (GTEC) under dry and humid conditions and different temperatures. Experiments are also designed to probe different peroxy radical pathways (RO2+HO2 vs RO2+NO3). Speciated gas-phase and particle-phase organic nitrates are continuously monitored by a Filter Inlet for Gases and AEROsols High Resolution Time-of-Flight Chemical Ionization Mass Spectrometer (FIGAERO-HR-ToF-CIMS). Bulk aerosol composition is measured by a High Resolution Time-of-Flight Aerosol Mass Spectrometer (HR-ToF-AMS). A large suite of highly oxygenated gas- and particle-phase organic nitrates are formed rapidly. We find a resistance to aerosol evaporation when it is heated. The extent of organic nitrate hydrolysis in the humid experiments is evaluated. The dynamics of the speciated organic nitrates over the course of the experiments will also be discussed. Results from this chamber study provide fundamental data for understanding the dynamics of organic nitrate aerosols over its atmospheric lifetime.

Relative association of Rubisco with manganese and magnesium as a regulatory mechanism in plants.

PubMed

Bloom, Arnold J; Kameritsch, Petra

2017-12-01

Rubisco, the enzyme that constitutes as much as half of the protein in a leaf, initiates either the photorespiratory pathway that supplies reductant for the assimilation of nitrate into amino acids or the C3 carbon fixation pathway that generates carbohydrates. The relative rates of these two pathways depend both on the relative extent to which O 2 and CO 2 occupies the active site of Rubisco and on whether manganese or magnesium is bound to the enzyme. This study quantified the activities of manganese and magnesium in isolated tobacco chloroplasts and the thermodynamics of binding of these metals to Rubisco purified from tobacco or a bacterium. In tobacco chloroplasts, manganese was less active than magnesium, but Rubisco purified from tobacco had a higher affinity for manganese. The activity of each metal in the chloroplast was similar in magnitude to the affinity of tobacco Rubisco for each. This indicates that, in tobacco chloroplasts, Rubisco associates almost equally with both metals and rapidly exchanges one metal for the other. Binding of magnesium was similar in Rubisco from tobacco and a bacterium, whereas binding of manganese differed greatly between the Rubisco from these species. Moreover, the ratio of leaf manganese to magnesium in C3 plants increased as atmospheric CO 2 increased. These results suggest that Rubisco has evolved to improve the energy transfers between photorespiration and nitrate assimilation and that plants regulate manganese and magnesium activities in the chloroplast to mitigate detrimental changes in their nitrogen/carbon balance as atmospheric CO 2 varies. © 2017 Scandinavian Plant Physiology Society.

Determining Roles of Accessory Genes in Denitrification by Mutant Fitness Analyses

DOE PAGES

Vaccaro, Brian J.; Thorgersen, Michael P.; Lancaster, W. Andrew; ...

2015-10-09

Enzymes of the denitrification pathway play an important role in the global nitrogen cycle, including release of nitrous oxide, an ozone-depleting greenhouse gas. In addition, nitric oxide reductase, maturation factors, and proteins associated with nitric oxide detoxification are used by pathogens to combat nitric oxide release by host immune systems. While the core reductases that catalyze the conversion of nitrate to dinitrogen are well understood at a mechanistic level, there are many peripheral proteins required for denitrification whose basic function is unclear. A bar-coded transposon DNA library fromPseudomonas stutzeristrain RCH2 was grown under denitrifying conditions, using nitrate or nitrite asmore » an electron acceptor, and also under molybdenum limitation conditions, with nitrate as the electron acceptor. Analysis of sequencing results from these growths yielded gene fitness data for 3,307 of the 4,265 protein-encoding genes present in strain RCH2. The insights presented here contribute to our understanding of how peripheral proteins contribute to a fully functioning denitrification pathway. We propose a new low-affinity molybdate transporter, OatABC, and show that differential regulation is observed for two MoaA homologs involved in molybdenum cofactor biosynthesis. We also propose that NnrS may function as a membrane-bound NO sensor. Finally, the dominant HemN paralog involved in heme biosynthesis is identified, and a CheR homolog is proposed to function in nitrate chemotaxis. In addition, new insights are provided into nitrite reductase redundancy, nitric oxide reductase maturation, nitrous oxide reductase maturation, and regulation.« less

Vulnerability of streams to legacy nitrate sources

USGS Publications Warehouse

Tesoriero, Anthony J.; Duff, John H.; Saad, David A.; Spahr, Norman E.; Wolock, David M.

2013-01-01

The influence of hydrogeologic setting on the susceptibility of streams to legacy nitrate was examined at seven study sites having a wide range of base flow index (BFI) values. BFI is the ratio of base flow to total streamflow volume. The portion of annual stream nitrate loads from base flow was strongly correlated with BFI. Furthermore, dissolved oxygen concentrations in streambed pore water were significantly higher in high BFI watersheds than in low BFI watersheds suggesting that geochemical conditions favor nitrate transport through the bed when BFI is high. Results from a groundwater-surface water interaction study at a high BFI watershed indicate that decades old nitrate-laden water is discharging to this stream. These findings indicate that high nitrate levels in this stream may be sustained for decades to come regardless of current practices. It is hypothesized that a first approximation of stream vulnerability to legacy nutrients may be made by geospatial analysis of watersheds with high nitrogen inputs and a strong connection to groundwater (e.g., high BFI).

Characterization of the three different states of the cholecystokinin (CCK) receptor in pancreatic acini.

PubMed

Talkad, V D; Patto, R J; Metz, D C; Turner, R J; Fortune, K P; Bhat, S T; Gardner, J D

1994-10-20

By measuring binding of [125I]CCK-8 and [3H]L-364,718 to rat pancreatic acini we demonstrated directly that the pancreatic CCK receptor can exist in three different affinity states with respect to CCK--high affinity, low affinity and very low affinity. Binding of [125I]CCK-8 reflects interaction of the tracer with the high and low affinity states, whereas binding of [3H]L-364,718 reflects interaction of the tracer with the low and very low affinity states. Treating acini with carbachol abolished the high affinity state of the CCK receptor and converted approximately 25% of the low affinity receptors to the very low affinity state. Carbachol treatment was particularly useful in establishing the values of Kd for the high and low affinity states for different CCK receptor agonists and antagonists. Of the various CCK receptor agonists tested, CCK-8 had the highest affinity for the high affinity state (Kd approximately 1 nM), whereas CCK-JMV-180 had the highest affinity for the low (Kd 7 nM) and very low affinity (Kd 200 nM) states. Gastrin and de(SO4)CCK-8 had affinities for the high and low affinity states of the receptor that were 100- to 400-fold less than those of CCK-8 but had affinities for the very low affinity state that were only 3- to 10-fold less than that of CCK-8. CCK receptor antagonists showed several patterns in interacting with the different states of the CCK receptor. L-364,718 had the same affinity for each state of the CCK receptor. CR1409 and Bt2cGMP each had similar affinities for the high and low affinity states and lower affinity for the very low affinity state. L-365,260 and CCK-JMV-179 had the highest affinity for the low affinity state and lower affinities for the high and very low affinity states. Different CCK receptor agonists caused the same maximal stimulation of amylase secretion but showed different degrees of amplification in terms of the relationship between their abilities to stimulate amylase secretion and their abilities to occupy the low affinity state of the CCK receptor. When amplification was expressed quantitatively as the value of Kd for the low affinity state divided by the corresponding EC50 for stimulating amylase secretion the values were CCK-8 (1000), de(SO)CCK-8 (1500), gastrin (100) and CCK-JMV-180 (Menozzi, D., Vinayek, R., Jensen, R.T. and Gardner, J.D. (1991) J. Biol. Chem. 266, 10385-1091).(ABSTRACT TRUNCATED AT 400 WORDS)

Totomatix: a novel automatic set-up to control diurnal, diel and long-term plant nitrate nutrition

PubMed Central

Adamowicz, Stéphane; Le Bot, Jacques; Huanosto Magaña, Ruth; Fabre, José

2012-01-01

Background Stand-alone nutritional set-ups are useful tools to grow plants at defined nutrient availabilities and to measure nutrient uptake rates continuously, in particular that for nitrate. Their use is essential when the measurements are meant to cover long time periods. These complex systems have, however, important drawbacks, including poor long-term reliability and low precision at high nitrate concentration. This explains why the information dealing with diel dynamics of nitrate uptake rate is scarce and concerns mainly young plants grown at low nitrate concentration. Scope The novel system detailed in this paper has been developed to allow versatile use in growth rooms, greenhouses or open fields at nitrate concentrations ranging from a few micro- to several millimoles per litres. The system controls, at set frequencies, the solution nitrate concentration, pH and volumes. Nitrate concentration is measured by spectral deconvolution of UV spectra. The main advantages of the set-up are its low maintenance (weekly basis), an ability to diagnose interference or erroneous analyses and high precision of nitrate concentration measurements (0·025 % at 3 mm). The paper details the precision of diurnal nitrate uptake rate measurements, which reveals sensitivity to solution volume at low nitrate concentration, whereas at high concentration, it is mostly sensitive to the precision of volume estimates. Conclusions This novel set-up allows us to measure and characterize the dynamics of plant nitrate nutrition at high temporal resolution (minutes to hours) over long-term experiments (up to 1 year). It is reliable and also offers a novel method to regulate up to seven N treatments by adjusting the daily uptake of test plants relative to controls, in variable environments such as open fields and glasshouses. PMID:21985796

Real time in situ detection of organic nitrates in atmospheric aerosols.

PubMed

Rollins, Andrew W; Smith, Jared D; Wilson, Kevin R; Cohen, Ronald C

2010-07-15

A novel instrument is described that quantifies total particle-phase organic nitrates in real time with a detection limit of 0.11 microg m(-3) min(-1), 45 ppt min(-1) (-ONO(2)). Aerosol nitrates are separated from gas-phase nitrates with a short residence time activated carbon denuder. Detection of organic molecules containing -ONO(2) subunits is accomplished using thermal dissociation coupled to laser induced fluorescence detection of NO(2). This instrument is capable of high time resolution (seconds) measurements of particle-phase organic nitrates, without interference from inorganic nitrate. Here we use it to quantify organic nitrates in secondary organic aerosol generated from high-NO(x) photooxidation of limonene, alpha-pinene, Delta-3-carene, and tridecane. In these experiments the organic nitrate moiety is observed to be 6-15% of the total SOA mass.

Quantifying watershed-scale groundwater loading and in-stream fate of nitrate using high-frequency water quality data

USGS Publications Warehouse

Miller, Matthew P.; Tesoriero, Anthony J.; Capel, Paul D.; Pellerin, Brian A.; Hyer, Kenneth E.; Burns, Douglas A.

2016-01-01

We describe a new approach that couples hydrograph separation with high-frequency nitrate data to quantify time-variable groundwater and runoff loading of nitrate to streams, and the net in-stream fate of nitrate at the watershed-scale. The approach was applied at three sites spanning gradients in watershed size and land use in the Chesapeake Bay watershed. Results indicate that 58-73% of the annual nitrate load to the streams was groundwater-discharged nitrate. Average annual first order nitrate loss rate constants (k) were similar to those reported in both modelling and in-stream process-based studies, and were greater at the small streams (0.06 and 0.22 d-1) than at the large river (0.05 d-1), but 11% of the annual loads were retained/lost in the small streams, compared with 23% in the large river. Larger streambed area to water volume ratios in small streams result in greater loss rates, but shorter residence times in small streams result in a smaller fraction of nitrate loads being removed than in larger streams. A seasonal evaluation of k values suggests that nitrate was retained/lost at varying rates during the growing season. Consistent with previous studies, streamflow and nitrate concentration were inversely related to k. This new approach for interpreting high-frequency nitrate data and the associated findings furthers our ability to understand, predict, and mitigate nitrate impacts on streams and receiving waters by providing insights into temporal nitrate dynamics that would be difficult to obtain using traditional field-based studies.

Determination of the origin of groundwater nitrate at an air weapons range using the dual isotope approach.

PubMed

Bordeleau, Geneviève; Savard, Martine M; Martel, Richard; Ampleman, Guy; Thiboutot, Sonia

2008-06-06

Nitrate is one of the most common contaminants in shallow groundwater, and many sources may contribute to the nitrate load within an aquifer. Groundwater nitrate plumes have been detected at several ammunition production sites. However, the presence of multiple potential sources and the lack of existing isotopic data concerning explosive degradation-induced nitrate constitute a limitation when it comes to linking both types of contaminants. On military training ranges, high nitrate concentrations in groundwater were reported for the first time as part of the hydrogeological characterization of the Cold Lake Air Weapons Range (CLAWR), Alberta, Canada. Explosives degradation is thought to be the main source of nitrate contamination at CLAWR, as no other major source is present. Isotopic analyses of N and O in nitrate were performed on groundwater samples from the unconfined and confined aquifers; the dual isotopic analysis approach was used in order to increase the chances of identifying the source of nitrate. The isotopic ratios for the groundwater samples with low nitrate concentration suggested a natural origin with a strong contribution of anthropogenic atmospheric NOx. For the samples with nitrate concentration above the expected background level the isotopic ratios did not correspond to any source documented in the literature. Dissolved RDX samples were degraded in the laboratory and results showed that all reproduced degradation processes released nitrate with a strong fractionation. Laboratory isotopic values for RDX-derived NO(3)(-) produced a trend of high delta(18)O-low delta(15)N to low delta(18)O-high delta(15)N, and groundwater samples with nitrate concentrations above the expected background level appeared along this trend. Our results thus point toward a characteristic field of isotopic ratios for nitrate being derived from the degradation of RDX.

Nitrate ammonification in mangrove soils: a hidden source of nitrite?

PubMed Central

Balk, Melike; Laverman, Anniet M.; Keuskamp, Joost A.; Laanbroek, Hendrikus J.

2015-01-01

Nitrate reduction is considered to be a minor microbial pathway in the oxidation of mangrove-derived organic matter due to a limited supply of nitrate in mangrove soils. At a limited availability of this electron acceptor compared to the supply of degradable carbon, nitrate ammonification is thought to be the preferential pathway of nitrate reduction. Mangrove forest mutually differ in their productivity, which may lead to different available carbon to nitrate ratios in their soil. Hence, nitrate ammonification is expected to be of more importance in high- compared to low-productive forests. The hypothesis was tested in flow-through reactors that contain undisturbed mangrove soils from high-productive Avicennia germinans and Rhizophora mangle forests in Florida and low-productive Avicennia marina forests in Saudi Arabia. Nitrate was undetectable in the soils from both regions. It was assumed that a legacy of nitrate ammonification would be reflected by a higher ammonium production from these soils upon the addition of nitrate. Unexpectedly, the soils from the low-productive forests in Saudi Arabia produced considerably more ammonium than the soils from the high-productive forests in Florida. Hence, other environmental factors than productivity must govern the selection of nitrate ammonification or denitrification. A rather intriguing observation was the 1:1 production of nitrite and ammonium during the consumption of nitrate, more or less independent from sampling region, location, sampling depth, mangrove species and from the absence or presence of additional degradable carbon. This 1:1 ratio points to a coupled production of ammonium and nitrite by one group of nitrate-reducing microorganisms. Such a production of nitrite will be hidden by the presence of active nitrite-reducing microorganisms under the nitrate-limited conditions of most mangrove forest soils. PMID:25784903

Nitrate ammonification in mangrove soils: a hidden source of nitrite?

PubMed

Balk, Melike; Laverman, Anniet M; Keuskamp, Joost A; Laanbroek, Hendrikus J

2015-01-01

Nitrate reduction is considered to be a minor microbial pathway in the oxidation of mangrove-derived organic matter due to a limited supply of nitrate in mangrove soils. At a limited availability of this electron acceptor compared to the supply of degradable carbon, nitrate ammonification is thought to be the preferential pathway of nitrate reduction. Mangrove forest mutually differ in their productivity, which may lead to different available carbon to nitrate ratios in their soil. Hence, nitrate ammonification is expected to be of more importance in high- compared to low-productive forests. The hypothesis was tested in flow-through reactors that contain undisturbed mangrove soils from high-productive Avicennia germinans and Rhizophora mangle forests in Florida and low-productive Avicennia marina forests in Saudi Arabia. Nitrate was undetectable in the soils from both regions. It was assumed that a legacy of nitrate ammonification would be reflected by a higher ammonium production from these soils upon the addition of nitrate. Unexpectedly, the soils from the low-productive forests in Saudi Arabia produced considerably more ammonium than the soils from the high-productive forests in Florida. Hence, other environmental factors than productivity must govern the selection of nitrate ammonification or denitrification. A rather intriguing observation was the 1:1 production of nitrite and ammonium during the consumption of nitrate, more or less independent from sampling region, location, sampling depth, mangrove species and from the absence or presence of additional degradable carbon. This 1:1 ratio points to a coupled production of ammonium and nitrite by one group of nitrate-reducing microorganisms. Such a production of nitrite will be hidden by the presence of active nitrite-reducing microorganisms under the nitrate-limited conditions of most mangrove forest soils.

Nitrate removal from high strength nitrate-bearing wastes in granular sludge sequencing batch reactors.

PubMed

Krishna Mohan, Tulasi Venkata; Renu, Kadali; Nancharaiah, Yarlagadda Venkata; Satya Sai, Pedapati Murali; Venugopalan, Vayalam Purath

2016-02-01

A 6-L sequencing batch reactor (SBR) was operated for development of granular sludge capable of denitrification of high strength nitrates. Complete and stable denitrification of up to 5420 mg L(-1) nitrate-N (2710 mg L(-1) nitrate-N in reactor) was achieved by feeding simulated nitrate waste at a C/N ratio of 3. Compact and dense denitrifying granular sludge with relatively stable microbial community was developed during reactor operation. Accumulation of large amounts of nitrite due to incomplete denitrification occurred when the SBR was fed with 5420 mg L(-1) NO3-N at a C/N ratio of 2. Complete denitrification could not be achieved at this C/N ratio, even after one week of reactor operation as the nitrite levels continued to accumulate. In order to improve denitrification performance, the reactor was fed with nitrate concentrations of 1354 mg L(-1), while keeping C/N ratio at 2. Subsequently, nitrate concentration in the feed was increased in a step-wise manner to establish complete denitrification of 5420 mg L(-1) NO3-N at a C/N ratio of 2. The results show that substrate concentration plays an important role in denitrification of high strength nitrate by influencing nitrite accumulation. Complete denitrification of high strength nitrates can be achieved at lower substrate concentrations, by an appropriate acclimatization strategy. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Isotopic Evidence of Nitrate Sources and its Relationship to Algae in the San Joaquin River, California

NASA Astrophysics Data System (ADS)

Silva, S. R.; Kendall, C.; Young, M. B.; Stringfellow, W. T.; Borglin, S. E.; Kratzer, C. R.; Dahlgren, R. A.; Schmidt, C.; Rollog, M. E.

2007-12-01

Many competing demands have been placed on the San Joaquin River including deep water shipping, use as agricultural and drinking water, transport of agricultural and urban runoff, and recreation. These long-established demands limit the management options and increase the importance of understanding the river dynamics. The relationships among sources of water, nitrate, and algae in the San Joaquin River must be understood before management decisions can be made to optimize aquatic health. Isotopic analyses of water samples collected along the San Joaquin River in 2005-2007 have proven useful in assessing these relationships: sources of nitrate, the productivity of the San Joaquin River, and the relationship between nitrate and algae in the river. The San Joaquin River receives water locally from wetlands and agricultural return flow, and from three relatively large tributaries whose headwaters are in the Sierra Nevada. The lowest nitrate concentrations occur during periods of high flow when the proportion of water from the Sierra Nevada is relatively large, reflecting the effect of dilution from the big tributaries and indicating that a large fraction of the nitrate is of local origin. Nitrogen isotopes of nitrate in the San Joaquin River are relatively high (averaging about 12 per mil), suggesting a significant source from animal waste or sewage and/or the effects of denitrification. The d15N of nitrate varies inversely with concentration, indicating that these high isotopic values are also a local product. The d15N values of nitrate from most of the local tributaries is lower than that in the San Joaquin suggesting that nitrate from these tributaries does not account for a significant fraction of nitrate in the river. The source of the non-tributary nitrate must be either small unmeasured surface inputs or groundwater. To investigate whether groundwater might be a significant source of nitrate to the San Joaquin River, groundwater samples are being collected monthly from over 20 bank and in-stream wells. Preliminary data suggest that much of the groundwater nitrate has been variably denitrified thereby increasing its d15N values, but not by enough to account for the high d15N values in the river nitrate. The d15N of algae in the San Joaquin reflects the high values of the nitrate in the river indicating (1) that the San Joaquin is productive despite its relatively high opacity, (2) that the algae use the nitrate as a primary nutrient source, and (3) that the concentrations of algae in the San Joaquin are not principally dependent on algae from the tributaries being flushed into the river as has been suggested. The sources of nitrate to the San Joaquin River must be identified if algae production is to be controlled and hypoxic conditions in the downstream reaches eliminated.

An alternative approach for nitrate and arsenic removal from wastewater via a nitrate-dependent ferrous oxidation process.

PubMed

Zhang, Meilin; Li, Yingfen; Long, Xinxian; Chong, Yunxiao; Yu, Guangwei; He, Zihao

2018-05-18

Owing to the high efficiency of converting nitrate to nitrogen gas with ferrous iron as the electron donor, the process of nitrate-dependent ferrous oxidation (NDFeO) has been considered suitable to treat wastewater that contains nitrate but lacks organic matter. Meanwhile, arsenic immobilization often has been found during the NDFeO reaction. Thus, it was strongly expected that nitrate and arsenic could be removed simultaneously in co-contaminated wastewater through the NDFeO process. However, in the current work, arsenic was not removed during the NDFeO process when the pH was high (above 8), though the nitrate reduction rate was over 90%. Meanwhile, the biosolid particles from the NDFeO process demonstrated strong adsorption ability for arsenic when the pH was below 6. Yet, the adsorption became weak when the pH was above 7. Fourier transform infrared (FTIR) spectroscopy analysis revealed that the main activated component for arsenic adsorption was iron oxide in these particles, which was easily crippled under high pH conditions. These results implied that co-removal of nitrate and arsenic in wastewater treatment using NDFeO was difficult to carry out under high pH conditions. Thus, a two-step approach in which nitrate was removed first by NDFeO followed by arsenic adsorption with NDFeO biosolids was more feasible. Copyright © 2018 Elsevier Ltd. All rights reserved.

Differential Effect of Irradiance and Nutrient Nitrate on the Relationship of in Vivo and in Vitro Nitrate Reductase Assay in Chlorophyllous Tissues 1

PubMed Central

Jones, Richard Wyn; Sheard, Robert W.

1977-01-01

Growth at increasing continuous irradiance (at high nutrient nitrate) and nutrient nitrate concentrations (at high continuous irradiance) furnished increases in the in vivo and in vitro nitrate reductase activities of corn (Zea mays L.), field peas (Pisum arvense L.), wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), and globe amaranth (Gomphrena globosa L.) leaves and of marrow (Cucurbita pepo L.) cotyledons. Ratios of in vivo to in vitro activity declined exponentially in all species with increasing nitrate reductase levels promoted by nutrient nitrate. The ratios were more nearly independent of nitrate reductase levels generated by adjusting the irradiance; major exceptions were marrow and wheat at low (1.5 klux and less) irradiances and peas throughout the irradiance range, where decreases in the ratio were accompanied by increases in in situ nitrate concentration. The ratio also increased at the highest irradiance (39.2 klux) in wheat and barley, associated with a decline of in vitro nitrate reductase. These differences in response to irradiance and nutrient nitrate indicate that the in vivo assay does not provide a simple measure of nitrate reductase but rather yields a more composite measure of nitrate reduction, possibly related both to nitrate reductase level and to the supply of reductant for in vivo activity. PMID:16659888

SEDIMENT DENITRIFICATION IN THE YAQUINA ESTUARY, OREGON

EPA Science Inventory

Rivers draining watersheds of the Coast Range in the northwestern United States frequently contain high concentrations of dissolved nitrate, particularly after high flow events (up to 180 ?M nitrate-N). The nitrate source appears to originate from the fixation of atmospheric nit...

Investigation of processes leading to nitrate enrichment in soils in the Kalahari Region, Botswana

NASA Astrophysics Data System (ADS)

Schwiede, M.; Duijnisveld, W. H. M.; Böttcher, J.

In Southern Africa elevated nitrate concentrations are observed in mostly uninhabited semi-arid areas. In the Kalahari of Botswana groundwater locally exhibits concentrations up to 600 mg/l. It is assumed, that nitrate found in the groundwater originates mainly from nitrogen input and transformations in the soils. Our investigations in the Kalahari between Serowe and Orapa show that cattle raising is an important source for enhanced nitrate concentrations in the soils (Arenosols). But also in termite mounds very high nitrate stocks were found, and under natural vegetation (acacia trees and shrubs) nitrate concentrations were mostly unexpectedly high. This nitrate enrichment in the soils poses a serious threat to the groundwater quality. However, calculated soil water age distributions in the unsaturated zone clearly show that today’s nitrate pollution of the groundwater below the investigation area could originate from natural sources, but cannot be caused by the current land use for cattle raising.

Availability of phosphate for phytoplankton and bacteria and of glucose for bacteria at different pCO2 levels in a mesocosm study

NASA Astrophysics Data System (ADS)

Tanaka, T.; Thingstad, T. F.; Løvdal, T.; Grossart, H.-P.; Larsen, A.; Allgaier, M.; Meyerhöfer, M.; Schulz, K. G.; Wohlers, J.; Zöllner, E.; Riebesell, U.

2008-05-01

Availability of phosphate for phytoplankton and bacteria and of glucose for bacteria at different pCO2 levels were studied in a mesocosm experiment (PeECE III). Using nutrient-depleted SW Norwegian fjord waters, three different levels of pCO2 (350 μatm: 1×CO2; 700 μatm: 2×CO2; 1050 μatm: 3×CO2) were set up, and nitrate and phosphate were added at the start of the experiment in order to induce a phytoplankton bloom. Despite similar responses of total particulate P concentration and phosphate turnover time at the three different pCO2 levels, the size distribution of particulate P and 33PO4 uptake suggested that phosphate transferred to the >10 μm fraction was greater in the 3×CO2 mesocosm during the first 6-10 days when phosphate concentration was high. During the period of phosphate depletion (after Day 12), specific phosphate affinity and specific alkaline phosphatase activity (APA) suggested a P-deficiency (i.e. suboptimal phosphate supply) rather than a P-limitation for the phytoplankton and bacterial community at the three different pCO2 levels. Specific phosphate affinity and specific APA tended to be higher in the 3×CO2 than in the 2×CO2 and 1×CO2 mesocosms during the phosphate depletion period, although no statistical differences were found. Glucose turnover time was correlated significantly and negatively with bacterial abundance and production but not with the bulk DOC concentration. This suggests that even though constituting a small fraction of the bulk DOC, glucose was an important component of labile DOC for bacteria. Specific glucose affinity of bacteria behaved similarly at the three different pCO2 levels with measured specific glucose affinities being consistently much lower than the theoretical maximum predicted from the diffusion-limited model. This suggests that bacterial growth was not severely limited by the glucose availability. Hence, it seems that the lower availability of inorganic nutrients after the phytoplankton bloom reduced the bacterial capacity to consume labile DOC in the upper mixed layer of the stratified mesocosms.

Food sources of nitrates and nitrites: the physiologic context for potential health benefits.

PubMed

Hord, Norman G; Tang, Yaoping; Bryan, Nathan S

2009-07-01

The presence of nitrates and nitrites in food is associated with an increased risk of gastrointestinal cancer and, in infants, methemoglobinemia. Despite the physiologic roles for nitrate and nitrite in vascular and immune function, consideration of food sources of nitrates and nitrites as healthful dietary components has received little attention. Approximately 80% of dietary nitrates are derived from vegetable consumption; sources of nitrites include vegetables, fruit, and processed meats. Nitrites are produced endogenously through the oxidation of nitric oxide and through a reduction of nitrate by commensal bacteria in the mouth and gastrointestinal tract. As such, the dietary provision of nitrates and nitrites from vegetables and fruit may contribute to the blood pressure-lowering effects of the Dietary Approaches to Stop Hypertension (DASH) diet. We quantified nitrate and nitrite concentrations by HPLC in a convenience sample of foods. Incorporating these values into 2 hypothetical dietary patterns that emphasize high-nitrate or low-nitrate vegetable and fruit choices based on the DASH diet, we found that nitrate concentrations in these 2 patterns vary from 174 to 1222 mg. The hypothetical high-nitrate DASH diet pattern exceeds the World Health Organization's Acceptable Daily Intake for nitrate by 550% for a 60-kg adult. These data call into question the rationale for recommendations to limit nitrate and nitrite consumption from plant foods; a comprehensive reevaluation of the health effects of food sources of nitrates and nitrites is appropriate. The strength of the evidence linking the consumption of nitrate- and nitrite-containing plant foods to beneficial health effects supports the consideration of these compounds as nutrients.

Insulin enhances the peroxidase activity of heme by forming heme-insulin complex: Relevance to type 2 diabetes mellitus.

PubMed

Huang, Yi; Yang, Zhen; Xu, Huan; Zhang, Pengfei; Gao, Zhonghong; Li, Hailing

2017-09-01

Evidences have implicated the involvement of heme in the type 2 diabetes mellitus (T2Dm) pathogenesis, but possible mediators linking between heme and diabetes are still poorly understood. Here, we explored a potential mechanism that linked heme, insulin and diabetes. Our results demonstrated the formation of heme-insulin complex by two classical methods, i.e. UV-vis and capillary electrophoresis-frontal analysis (CE-FA). UV-vis results implied heme binding insulin via bis-histidine sites, and CE-FA further revealed that, when insulin uses two sites binding with heme, this interaction occurs at high affinity (K d =3.13×10 -6 M). Molecule docking supported that histidine-B5 of insulin binds with heme-Fe. In addition to that, tyrosine-B26, phenylalanine-B1 and valine-B2 are also contributed to binding heme. The binding amplified the peroxidase activity of heme itself. Under oxidative and nitrative stress, it affects pathogenesis of diabetes from two aspects: promoting insulin cross-linking that leads to permanent loss of insulin functionality on one hand, and enhancing protein tyrosine nitration that may result in inactivation of proteins associated with diabetes on the other hand. This study suggested that the enhanced peroxidase activity of heme through binding with insulin might be a previously unrecognized contributor to the pathogenesis of T2Dm in some heme-associated disorders. Copyright © 2017 Elsevier B.V. All rights reserved.

Arabidopsis roots and shoots show distinct temporal adaptation patterns toward nitrogen starvation.

PubMed

Krapp, Anne; Berthomé, Richard; Orsel, Mathilde; Mercey-Boutet, Stéphanie; Yu, Agnes; Castaings, Loren; Elftieh, Samira; Major, Hilary; Renou, Jean-Pierre; Daniel-Vedele, Françoise

2011-11-01

Nitrogen (N) is an essential macronutrient for plants. N levels in soil vary widely, and plants have developed strategies to cope with N deficiency. However, the regulation of these adaptive responses and the coordinating signals that underlie them are still poorly understood. The aim of this study was to characterize N starvation in adult Arabidopsis (Arabidopsis thaliana) plants in a spatiotemporal manner by an integrative, multilevel global approach analyzing growth, metabolites, enzyme activities, and transcript levels. We determined that the remobilization of N and carbon compounds to the growing roots occurred long before the internal N stores became depleted. A global metabolite analysis by gas chromatography-mass spectrometry revealed organ-specific differences in the metabolic adaptation to complete N starvation, for example, for several tricarboxylic acid cycle intermediates, but also for carbohydrates, secondary products, and phosphate. The activities of central N metabolism enzymes and the capacity for nitrate uptake adapted to N starvation by favoring N remobilization and by increasing the high-affinity nitrate uptake capacity after long-term starvation. Changes in the transcriptome confirmed earlier studies and added a new dimension by revealing specific spatiotemporal patterns and several unknown N starvation-regulated genes, including new predicted small RNA genes. No global correlation between metabolites, enzyme activities, and transcripts was evident. However, this multilevel spatiotemporal global study revealed numerous new patterns of adaptation mechanisms to N starvation. In the context of a sustainable agriculture, this work will give new insight for the production of crops with increased N use efficiency.

Control of Sulfide Production in High Salinity Bakken Shale Oil Reservoirs by Halophilic Bacteria Reducing Nitrate to Nitrite.

PubMed

An, Biwen A; Shen, Yin; Voordouw, Gerrit

2017-01-01

Microbial communities in shale oil fields are still poorly known. We obtained samples of injection, produced and facility waters from a Bakken shale oil field in Saskatchewan, Canada with a resident temperature of 60°C. The injection water had a lower salinity (0.7 Meq of NaCl) than produced or facility waters (0.6-3.6 Meq of NaCl). Salinities of the latter decreased with time, likely due to injection of low salinity water, which had 15-30 mM sulfate. Batch cultures of field samples showed sulfate-reducing and nitrate-reducing bacteria activities at different salinities (0, 0.5, 0.75, 1.0, 1.5, and 2.5 M NaCl). Notably, at high salinity nitrite accumulated, which was not observed at low salinity, indicating potential for nitrate-mediated souring control at high salinity. Continuous culture chemostats were established in media with volatile fatty acids (a mixture of acetate, propionate and butyrate) or lactate as electron donor and nitrate or sulfate as electron acceptor at 0.5 to 2.5 M NaCl. Microbial community analyses of these cultures indicated high proportions of Halanaerobium, Desulfovermiculus, Halomonas , and Marinobacter in cultures at 2.5 M NaCl, whereas Desulfovibrio, Geoalkalibacter , and Dethiosulfatibacter were dominant at 0.5 M NaCl. Use of bioreactors to study the effect of nitrate injection on sulfate reduction showed that accumulation of nitrite inhibited SRB activity at 2.5 M but not at 0.5 M NaCl. High proportions of Halanaerobium and Desulfovermiculus were found at 2.5 M NaCl in the absence of nitrate, whereas high proportions of Halomonas and no SRB were found in the presence of nitrate. A diverse microbial community dominated by the SRB Desulfovibrio was observed at 0.5 M NaCl both in the presence and absence of nitrate. Our results suggest that nitrate injection can prevent souring provided that the salinity is maintained at a high level. Thus, reinjection of high salinity produced water amended with nitrate maybe be a cost effective method for souring control.

Control of Sulfide Production in High Salinity Bakken Shale Oil Reservoirs by Halophilic Bacteria Reducing Nitrate to Nitrite

PubMed Central

An, Biwen A.; Shen, Yin; Voordouw, Gerrit

2017-01-01

Microbial communities in shale oil fields are still poorly known. We obtained samples of injection, produced and facility waters from a Bakken shale oil field in Saskatchewan, Canada with a resident temperature of 60°C. The injection water had a lower salinity (0.7 Meq of NaCl) than produced or facility waters (0.6–3.6 Meq of NaCl). Salinities of the latter decreased with time, likely due to injection of low salinity water, which had 15–30 mM sulfate. Batch cultures of field samples showed sulfate-reducing and nitrate-reducing bacteria activities at different salinities (0, 0.5, 0.75, 1.0, 1.5, and 2.5 M NaCl). Notably, at high salinity nitrite accumulated, which was not observed at low salinity, indicating potential for nitrate-mediated souring control at high salinity. Continuous culture chemostats were established in media with volatile fatty acids (a mixture of acetate, propionate and butyrate) or lactate as electron donor and nitrate or sulfate as electron acceptor at 0.5 to 2.5 M NaCl. Microbial community analyses of these cultures indicated high proportions of Halanaerobium, Desulfovermiculus, Halomonas, and Marinobacter in cultures at 2.5 M NaCl, whereas Desulfovibrio, Geoalkalibacter, and Dethiosulfatibacter were dominant at 0.5 M NaCl. Use of bioreactors to study the effect of nitrate injection on sulfate reduction showed that accumulation of nitrite inhibited SRB activity at 2.5 M but not at 0.5 M NaCl. High proportions of Halanaerobium and Desulfovermiculus were found at 2.5 M NaCl in the absence of nitrate, whereas high proportions of Halomonas and no SRB were found in the presence of nitrate. A diverse microbial community dominated by the SRB Desulfovibrio was observed at 0.5 M NaCl both in the presence and absence of nitrate. Our results suggest that nitrate injection can prevent souring provided that the salinity is maintained at a high level. Thus, reinjection of high salinity produced water amended with nitrate maybe be a cost effective method for souring control. PMID:28680423

Nitrate-driven urban haze pollution during summertime over the North China Plain

NASA Astrophysics Data System (ADS)

Li, Haiyan; Zhang, Qiang; Zheng, Bo; Chen, Chunrong; Wu, Nana; Guo, Hongyu; Zhang, Yuxuan; Zheng, Yixuan; Li, Xin; He, Kebin

2018-04-01

Compared to the severe winter haze episodes in the North China Plain (NCP), haze pollution during summertime has drawn little public attention. In this study, we present the highly time-resolved chemical composition of submicron particles (PM1) measured in Beijing and Xinxiang in the NCP region during summertime to evaluate the driving factors of aerosol pollution. During the campaign periods (30 June to 27 July 2015, for Beijing and 8 to 25 June 2017, for Xinxiang), the average PM1 concentrations were 35.0 and 64.2 µg m-3 in Beijing and Xinxiang. Pollution episodes characterized with largely enhanced nitrate concentrations were observed at both sites. In contrast to the slightly decreased mass fractions of sulfate, semivolatile oxygenated organic aerosol (SV-OOA), and low-volatility oxygenated organic aerosol (LV-OOA) in PM1, nitrate displayed a significantly enhanced contribution with the aggravation of aerosol pollution, highlighting the importance of nitrate formation as the driving force of haze evolution in summer. Rapid nitrate production mainly occurred after midnight, with a higher formation rate than that of sulfate, SV-OOA, or LV-OOA. Based on observation measurements and thermodynamic modeling, high ammonia emissions in the NCP region favored the high nitrate production in summer. Nighttime nitrate formation through heterogeneous hydrolysis of dinitrogen pentoxide (N2O5) enhanced with the development of haze pollution. In addition, air masses from surrounding polluted areas during haze episodes led to more nitrate production. Finally, atmospheric particulate nitrate data acquired by mass spectrometric techniques from various field campaigns in Asia, Europe, and North America uncovered a higher concentration and higher fraction of nitrate present in China. Although measurements in Beijing during different years demonstrate a decline in the nitrate concentration in recent years, the nitrate contribution in PM1 still remains high. To effectively alleviate particulate matter pollution in summer, our results suggest an urgent need to initiate ammonia emission control measures and further reduce nitrogen oxide emissions over the NCP region.

Myeloperoxidase-generated reactive nitrogen species convert LDL into an atherogenic form in vitro

PubMed Central

Podrez, Eugene A.; Schmitt, David; Hoff, Henry F.; Hazen, Stanley L.

1999-01-01

Oxidized LDL is implicated in atherosclerosis; however, the pathways that convert LDL into an atherogenic form in vivo are not established. Production of reactive nitrogen species may be one important pathway, since LDL recovered from human atherosclerotic aorta is enriched in nitrotyrosine. We now report that reactive nitrogen species generated by the MPO-H2O2-NO2– system of monocytes convert LDL into a form (NO2-LDL) that is avidly taken up and degraded by macrophages, leading to massive cholesterol deposition and foam cell formation, essential steps in lesion development. Incubation of LDL with isolated MPO, an H2O2-generating system, and nitrite (NO2–)— a major end-product of NO metabolism—resulted in nitration of apolipoprotein B 100 tyrosyl residues and initiation of LDL lipid peroxidation. The time course of LDL protein nitration and lipid peroxidation paralleled the acquisition of high-affinity, concentration-dependent, and saturable binding of NO2-LDL to human monocyte–derived macrophages and mouse peritoneal macrophages. LDL modification and conversion into a high-uptake form occurred in the absence of free metal ions, required NO2–, occurred at physiological levels of Cl–, and was inhibited by heme poisons, catalase, and BHT. Macrophage binding of NO2-LDL was specific and mediated by neither the LDL receptor nor the scavenger receptor class A type I. Exposure of macrophages to NO2-LDL promoted cholesteryl ester synthesis, intracellular cholesterol and cholesteryl ester accumulation, and foam cell formation. Collectively, these results identify MPO-generated reactive nitrogen species as a physiologically plausible pathway for converting LDL into an atherogenic form. PMID:10359564

Cloning and functional characterization of the high-affinity K+ transporter HAK1 of pepper.

PubMed

Martínez-Cordero, M Angeles; Martínez, Vicente; Rubio, Francisco

2004-10-01

High-affinity K+ uptake in plants plays a crucial role in K+ nutrition and different systems have been postulated to contribute to the high-affinity K+ uptake. The results presented here with pepper (Capsicum annum) demonstrate that a HAK1-type transporter greatly contributes to the high-affinity K+ uptake observed in roots. Pepper plants starved of K+ for 3 d showed high-affinity K+ uptake (Km of 6 microM K+) that was very sensitive to NH and their roots expressed a high-affinity K+ transporter, CaHAK1, which clusters in group I of the KT/HAK/KUP family of transporters. When expressed in yeast ( Saccharomyces cerevisiae ), CaHAK1 mediated high-affinity K+ and Rb+ uptake with Km values of 3.3 and 1.9 microM, respectively. Rb+ uptake was competitively inhibited by micromolar concentrations of NH and Cs+, and by millimolar concentrations of Na+.

Nitrate Loads and Concentrations in Surface-Water Base Flow and Shallow Groundwater for Selected Basins in the United States, Water Years 1990-2006

USGS Publications Warehouse

Spahr, Norman E.; Dubrovsky, Neil M.; Gronberg, JoAnn M.; Franke, O. Lehn; Wolock, David M.

2010-01-01

Hydrograph separation was used to determine the base-flow component of streamflow for 148 sites sampled as part of the National Water-Quality Assessment program. Sites in the Southwest and the Northwest tend to have base-flow index values greater than 0.5. Sites in the Midwest and the eastern portion of the Southern Plains generally have values less than 0.5. Base-flow index values for sites in the Southeast and Northeast are mixed with values less than and greater than 0.5. Hypothesized flow paths based on relative scaling of soil and bedrock permeability explain some of the differences found in base-flow index. Sites in areas with impermeable soils and bedrock (areas where overland flow may be the primary hydrologic flow path) tend to have lower base-flow index values than sites in areas with either permeable bedrock or permeable soils (areas where deep groundwater flow paths or shallow groundwater flow paths may occur). The percentage of nitrate load contributed by base flow was determined using total flow and base flow nitrate load models. These regression-based models were calibrated using available nitrate samples and total streamflow or base-flow nitrate samples and the base-flow component of total streamflow. Many streams in the country have a large proportion of nitrate load contributed by base flow: 40 percent of sites have more than 50 percent of the total nitrate load contributed by base flow. Sites in the Midwest and eastern portion of the Southern Plains generally have less than 50 percent of the total nitrate load contributed by base flow. Sites in the Northern Plains and Northwest have nitrate load ratios that generally are greater than 50 percent. Nitrate load ratios for sites in the Southeast and Northeast are mixed with values less than and greater than 50 percent. Significantly lower contributions of nitrate from base flow were found at sites in areas with impermeable soils and impermeable bedrock. These areas could be most responsive to nutrient management practices designed to reduce nutrient transport to streams by runoff. Conversely, sites with potential for shallow or deep groundwater contribution (some combination of permeable soils or permeable bedrock) had significantly greater contributions of nitrate from base flow. Effective nutrient management strategies would consider groundwater nitrate contributions in these areas. Mean annual base-flow nitrate concentrations were compared to shallow-groundwater nitrate concentrations for 27 sites. Concentrations in groundwater tended to be greater than base-flow concentrations for this group of sites. Sites where groundwater concentrations were much greater than base-flow concentrations were found in areas of high infiltration and oxic groundwater conditions. The lack of correspondingly high concentrations in the base flow of the paired surface-water sites may have multiple causes. In some settings, there has not been sufficient time for enough high-nitrate shallow groundwater to migrate to the nearby stream. In these cases, the stream nitrate concentrations lag behind those in the shallow groundwater, and concentrations may increase in the future as more high-nitrate groundwater reaches the stream. Alternatively, some of these sites may have processes that rapidly remove nitrate as water moves from the aquifer into the stream channel. Partitioning streamflow and nitrate load between the quick-flow and base-flow portions of the hydrograph coupled with relative scales of soil permeability can infer the importance of surface water compared to groundwater nitrate sources. Study of the relation of nitrate concentrations to base-flow index and the comparison of groundwater nitrate concentrations to stream nitrate concentrations during times when base-flow index is high can provide evidence of potential nitrate transport mechanisms. Accounting for the surface-water and groundwater contributions of nitrate is crucial to effective management and remediat

Knowledge discovery from high-frequency stream nitrate concentrations: hydrology and biology contributions.

PubMed

Aubert, Alice H; Thrun, Michael C; Breuer, Lutz; Ultsch, Alfred

2016-08-30

High-frequency, in-situ monitoring provides large environmental datasets. These datasets will likely bring new insights in landscape functioning and process scale understanding. However, tailoring data analysis methods is necessary. Here, we detach our analysis from the usual temporal analysis performed in hydrology to determine if it is possible to infer general rules regarding hydrochemistry from available large datasets. We combined a 2-year in-stream nitrate concentration time series (time resolution of 15 min) with concurrent hydrological, meteorological and soil moisture data. We removed the low-frequency variations through low-pass filtering, which suppressed seasonality. We then analyzed the high-frequency variability component using Pareto Density Estimation, which to our knowledge has not been applied to hydrology. The resulting distribution of nitrate concentrations revealed three normally distributed modes: low, medium and high. Studying the environmental conditions for each mode revealed the main control of nitrate concentration: the saturation state of the riparian zone. We found low nitrate concentrations under conditions of hydrological connectivity and dominant denitrifying biological processes, and we found high nitrate concentrations under hydrological recession conditions and dominant nitrifying biological processes. These results generalize our understanding of hydro-biogeochemical nitrate flux controls and bring useful information to the development of nitrogen process-based models at the landscape scale.

Skeletal muscle as an endogenous nitrate reservoir

PubMed Central

Piknova, Barbora; Park, Ji Won; Swanson, Kathryn M.; Dey, Soumyadeep; Noguchi, Constance Tom; Schechter, Alan N

2015-01-01

The nitric oxide synthase (NOS) family of enzymes form nitric oxide (NO) from arginine in the presence of oxygen. At reduced oxygen availability NO is also generated from nitrate in a two step process by bacterial and mammalian molybdopterin proteins, and also directly from nitrite by a variety of five-coordinated ferrous hemoproteins. The mammalian NO cycle also involves direct oxidation of NO to nitrite, and both NO and nitrite to nitrate by oxy-ferrous hemoproteins. The liver and blood are considered the sites of active mammalian NO metabolism and nitrite and nitrate concentrations in the liver and blood of several mammalian species, including human, have been determined. However, the large tissue mass of skeletal muscle had not been generally considered in the analysis of the NO cycle, in spite of its long-known presence of significant levels of active neuronal NOS (nNOS or NOS1). We hypothesized that skeletal muscle participates in the NO cycle and, due to its NO oxidizing heme protein, oxymyoglobin, has high concentrations of nitrate ions. We measured nitrite and nitrate concentrations in rat and mouse leg skeletal muscle and found unusually high concentrations of nitrate but similar levels of nitrite, when compared to the liver. The nitrate reservoir in muscle is easily accessible via the bloodstream and therefore nitrate is available for transport to internal organs where it can be reduced to nitrite and NO. Nitrate levels in skeletal muscle and blood in nNOS−/− mice were dramatically lower when compared with controls, which support further our hypothesis. Although the nitrate reductase activity of xanthine oxidoreductase in muscle is less than that of liver, the residual activity in muscle could be very important in view of its total mass and the high basal level of nitrate. We suggest that skeletal muscle participates in overall NO metabolism, serving as a nitrate reservoir, for direct formation of nitrite and NO, and for determining levels of nitrate in other organs. PMID:25727730

Isotopic Identification of Nitrate Sources and Cycling in Arctic Tundra Active Layer Soils and Permafrost

NASA Astrophysics Data System (ADS)

Heikoop, J. M.; Throckmorton, H.; Newman, B. D.; Perkins, G.; Gard, M.; Iversen, C. M.; Wilson, C. J.; Wullschleger, S. D.

2014-12-01

The effect of nitrogen cycling on release of carbon from tundra ecosystems is being studied as part of the US Department of Energy Next Generation Ecosystem Experiment - Arctic project. Sampling and analysis of active layer soil water at the Barrow Environmental Observatory (Alaska, USA) was performed in ancient drained thaw lake basins (DTLBs), drainages, and in polygonal terrain associated with inter-DTLB tundra. Within active layer soils, nitrate was most commonly found above analytical limits of detection in pore water from the unsaturated centers of high-centered polygons. Nitrate has also been detected, though less frequently, in soil water immediately above the frost table of an ancient (14C age of 2000 - 5500 BP) DTLB and in a small drainage adjacent to high-centered polygonal terrain. Nitrate from high-centered polygons had δ15N ranging from -9.2 to +8.5 ‰ and δ18O ranging from -8.4 to +1.4 ‰. The δ15N isotopic range is consistent with microbial mineralization and nitrification of reduced nitrogen sources including ammonium, dissolved organic nitrogen, and soil organic nitrogen. The range in δ18O of nitrate is also consistent with nitrification based on the δ18O of site waters. No evidence for an atmospheric nitrate signal, as defined by δ15N and δ18O of nitrate in snow and snowmelt, is seen. In contrast, nitrate in permafrost appears to be a mixture of pre-industrial atmospheric nitrate (with higher δ15N than modern atmospheric nitrate) and nitrate that is microbial in origin. Massive ice wedges appear to contain larger proportions of snowmelt (based on δ18O of ice) and atmospheric nitrate, whereas textural ice appears to contain a greater proportion of summer precipitation and microbially-derived nitrate. Nitrate from the ancient DTLB and drainage samples also has isotopic signatures that appear to represent a mixture of pre-industrial atmospheric nitrate and nitrate from microbial nitrification, and may, at least in part, be derived from degraded permafrost.

Simultaneous removal of cadmium and nitrate in aqueous media by nanoscale zerovalent iron (nZVI) and Au doped nZVI particles.

PubMed

Su, Yiming; Adeleye, Adeyemi S; Huang, Yuxiong; Sun, Xiaoya; Dai, Chaomeng; Zhou, Xuefei; Zhang, Yalei; Keller, Arturo A

2014-10-15

Nanoscale zerovalent iron (nZVI) has demonstrated high efficacy for treating nitrate or cadmium (Cd) contamination, but its efficiency for simultaneous removal of nitrate and Cd has not been investigated. This study evaluated the reactivity of nZVI to the co-contaminants and by-product formation, employed different catalysts to reduce nitrite yield from nitrate, and examined the transformation of nZVI after reaction. Nitrate reduction resulted in high solution pH, negatively charged surface of nZVI, formation of Fe3O4 (a stable transformation of nZVI), and no release of ionic iron. Increased pH and negative charge contributed to significant increase in Cd(II) removal capacity (from 40 mg/g to 188 mg/g) with nitrate present. In addition, nitrate reduction by nZVI could be catalyzed by Cd(II): while 30% of nitrate was reduced by nZVI within 2 h in the absence of Cd(II), complete nitrate reduction was observed in the presence of 40 mg-Cd/L due to the formation of Cd islands (Cd(0) and CdO) on the nZVI particles. While nitrate was reduced mostly to ammonium when Cd(II) was not present or at Cd(II) concentrations ≥ 40 mg/L, up to 20% of the initial nitrate was reduced to nitrite at Cd(II) concentrations < 40 mg/L. Among nZVI particles doped with 1 wt. % Cu, Ag, or Au, nZVI deposited with 1 wt. % Au reduced nitrite yield to less than 3% of the initial nitrate, while maintaining a high Cd(II) removal capacity. Copyright © 2014 Elsevier Ltd. All rights reserved.

Advanced Thermal Storage for Central Receivers with Supercritical Coolants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kelly, Bruce D.

2010-06-15

The principal objective of the study is to determine if supercritical heat transport fluids in a central receiver power plant, in combination with ceramic thermocline storage systems, offer a reduction in levelized energy cost over a baseline nitrate salt concept. The baseline concept uses a nitrate salt receiver, two-tank (hot and cold) nitrate salt thermal storage, and a subcritical Rankine cycle. A total of 6 plant designs were analyzed, as follows: Plant Designation Receiver Fluid Thermal Storage Rankine Cycle Subcritical nitrate salt Nitrate salt Two tank nitrate salt Subcritical Supercritical nitrate salt Nitrate salt Two tank nitrate salt Supercritical Lowmore » temperature H2O Supercritical H2O Two tank nitrate salt Supercritical High temperature H2O Supercritical H2O Packed bed thermocline Supercritical Low temperature CO2 Supercritical CO2 Two tank nitrate salt Supercritical High temperature CO2 Supercritical CO2 Packed bed thermocline Supercritical Several conclusions have been drawn from the results of the study, as follows: 1) The use of supercritical H2O as the heat transport fluid in a packed bed thermocline is likely not a practical approach. The specific heat of the fluid is a strong function of the temperatures at values near 400 °C, and the temperature profile in the bed during a charging cycle is markedly different than the profile during a discharging cycle. 2) The use of supercritical CO2 as the heat transport fluid in a packed bed thermocline is judged to be technically feasible. Nonetheless, the high operating pressures for the supercritical fluid require the use of pressure vessels to contain the storage inventory. The unit cost of the two-tank nitrate salt system is approximately $24/kWht, while the unit cost of the high pressure thermocline system is nominally 10 times as high. 3) For the supercritical fluids, the outer crown temperatures of the receiver tubes are in the range of 700 to 800 °C. At temperatures of 700 °C and above, intermetallic compounds can precipitate between, and within, the grains of nickel alloys. The precipitation leads to an increase in tensile strength, and a decrease in ductility. Whether the proposed tube materials can provide the required low cycle fatigue life for the supercritical H2O and CO2 receivers is an open question. 4) A ranking of the plants, in descending order of technical and economic feasibility, is as follows: i) Supercritical nitrate salt and baseline nitrate salt: equal ratings ii) Low temperature supercritical H2O iii) Low temperature supercritical CO2 iv) High temperature supercritical CO2 v) High temperature supercritical H2O 5) The two-tank nitrate salt thermal storage systems are strongly preferred over the thermocline systems using supercritical heat transport fluids.« less

High nitrate concentrations in some Midwest United States streams in 2013 after the 2012 drought

USGS Publications Warehouse

Van Metre, Peter C.; Frey, Jeffrey W.; Musgrove, MaryLynn; Nakagaki, Naomi; Qi, Sharon L.; Mahler, Barbara J.; Wieczorek, Michael; Button, Daniel T.

2016-01-01

Nitrogen sources in the Mississippi River basin have been linked to degradation of stream ecology and to Gulf of Mexico hypoxia. In 2013, the USGS and the USEPA characterized water quality stressors and ecological conditions in 100 wadeable streams across the midwestern United States. Wet conditions in 2013 followed a severe drought in 2012, a weather pattern associated with elevated nitrogen concentrations and loads in streams. Nitrate concentrations during the May to August 2013 sampling period ranged from <0.04 to 41.8 mg L−1 as N (mean, 5.31 mg L−1). Observed mean May to June nitrate concentrations at the 100 sites were compared with May to June concentrations predicted from a regression model developed using historical nitrate data. Observed concentrations for 17 sites, centered on Iowa and southern Minnesota, were outside the 95% confidence interval of the regression-predicted mean, indicating that they were anomalously high. The sites with a nitrate anomaly had significantly higher May to June nitrate concentrations than sites without an anomaly (means, 19.8 and 3.6 mg L−1, respectively) and had higher antecedent precipitation indices, a measure of the departure from normal precipitation, in 2012 and 2013. Correlations between nitrate concentrations and watershed characteristics and nitrogen and oxygen isotopes of nitrate indicated that fertilizer and manure used in crop production, principally corn, were the dominant sources of nitrate. The anomalously high nitrate levels in parts of the Midwest in 2013 coincide with reported higher-than-normal nitrate loads in the Mississippi River.

Antecedent flow conditions and nitrate concentrations in the Mississippi River basin

USGS Publications Warehouse

Murphy, Jennifer C.; Hirsch, Robert M.; Sprague, Lori A.

2014-01-01

The relationship between antecedent flow conditions and nitrate concentrations was explored at eight sites in the 2.9 million square kilometers (km2) Mississippi River basin, USA. Antecedent flow conditions were quantified as the ratio between the mean daily flow of the previous year and the mean daily flow from the period of record (Qratio), and the Qratio was statistically related to nitrate anomalies (the unexplained variability in nitrate concentration after filtering out season, long-term trend, and contemporaneous flow effects) at each site. Nitrate anomaly and Qratio were negatively related at three of the four major tributary sites and upstream in the Mississippi River, indicating that when mean daily streamflow during the previous year was lower than average, nitrate concentrations were higher than expected. The strength of these relationships increased when data were subdivided by contemporaneous flow conditions. Five of the eight sites had significant negative relationships (p ≤ 0.05) at high or moderately high contemporaneous flows, suggesting nitrate that accumulates in these basins during a drought is flushed during subsequent high flows. At half of the sites, when mean daily flow during the previous year was 50 percent lower than average, nitrate concentration can be from 9 to 27 percent higher than nitrate concentrations that follow a year with average mean daily flow. Conversely, nitrate concentration can be from 8 to 21 percent lower than expected when flow during the previous year was 50 percent higher than average. Previously documented for small, relatively homogenous basins, our results suggest that relationships between antecedent flows and nitrate concentrations are also observable at a regional scale. Relationships were not observed (using all contemporaneous flow data together) for basins larger than 1 million km2, suggesting that above this limit the overall size and diversity within these basins may necessitate the use of more complicated statistical approaches or that there may be no discernible basin-wide relationship with antecedent flow. The relationships between nitrate concentration and Qratio identified in this study serve as the basis for future studies that can better define specific hydrologic processes occurring during and after a drought (or high flow period) which influence nitrate concentration, such as the duration or magnitude of low flows, and the timing of low and high flows.

High-nitrate vegetable diet increases plasma nitrate and nitrite concentrations and reduces blood pressure in healthy women.

PubMed

Ashworth, Ann; Mitchell, Klaus; Blackwell, Jamie R; Vanhatalo, Anni; Jones, Andrew M

2015-10-01

Epidemiological studies suggest that green leafy vegetables, which are high in dietary nitrate, are protective against CVD such as stroke. High blood pressure (BP) is a major risk factor for stroke and inorganic nitrate has been shown to reduce BP. The objective of the present study was to test the hypothesis that diets containing high-nitrate (HN) vegetables would increase plasma nitrate and nitrite concentrations and reduce BP in healthy women. A randomized, crossover trial, where participants received HN vegetables (HN diet) or avoided HN vegetables (Control diet) for 1 week. Before and after each intervention, resting BP and plasma nitrate and nitrite concentrations were measured. University of Exeter, UK. Nineteen healthy women (mean age 20 (sd 2) years; mean BMI 22·5 (sd 3·8) kg/m2). The HN diet significantly increased plasma nitrate concentration (before HN diet: mean 24·4 (sd 5·6) µmol/l; after HN diet: mean 61·0 (sd 44·1) µmol/l, P<0·05) and plasma nitrite concentration (before HN diet: mean 98 (sd 91) nmol/l; after HN diet: mean 185 (sd 34) nmol/l, P<0·05). No significant change in plasma nitrate or nitrite concentration was observed after the Control diet. The HN diet significantly reduced resting systolic BP (before HN diet: mean 107 (sd 9) mmHg; after HN diet: mean 103 (sd 6) mmHg, P<0·05). No significant change in systolic BP was observed after the Control diet (before Control diet: mean 106 (sd 8) mmHg; after Control diet: mean 106 (sd 8) mmHg). Consumption of HN vegetables significantly increased plasma nitrate and nitrite concentrations and reduced BP in normotensive women.

Potential health consequences of ground-water contamination by nitrates in Nebraska.

PubMed

Weisenburger, D D

1993-01-01

Ground water serves as the primary source of drinking water for nearly all of rural Nebraska. However, ground-water contamination by nitrates, largely due to the use of fertilizers, is an increasing problem. In an ecologic study, the author found that counties characterized by high fertilizer usage and significant ground-water contamination by nitrates also had a high incidence of non-Hodgkin's lymphoma. Other potential health effects of nitrates in drinking water are also discussed.

A rapid and high-throughput microplate spectrophotometric method for field measurement of nitrate in seawater and freshwater.

PubMed

Wu, Jiapeng; Hong, Yiguo; Guan, Fengjie; Wang, Yan; Tan, Yehui; Yue, Weizhong; Wu, Meilin; Bin, Liying; Wang, Jiaping; Wen, Jiali

2016-02-01

The well-known zinc-cadmium reduction method is frequently used for determination of nitrate. However, this method is seldom to be applied on field research of nitrate due to the long time consuming and large sample volume demand. Here, we reported a modified zinc-cadmium reduction method (MZCRM) for measurement of nitrate at natural-abundance level in both seawater and freshwater. The main improvements of MZCRM include using small volume disposable tubes for reaction, a vortex apparatus for shaking to increase reduction rate, and a microplate reader for high-throughput spectrophotometric measurements. Considering salt effect, two salinity sections (5~10 psu and 20~35 psu) were set up for more accurate determination of nitrate in low and high salinity condition respectively. Under optimized experimental conditions, the reduction rates were stabilized on 72% and 63% on the salinity of 5 and 20 psu respectively. The lowest detection limit for nitrate was 0.5 μM and was linear up to 100 μM (RSDs was 4.8%). Environmental samples assay demonstrated that MZCRM was well consistent with conventional zinc-cadmium reduction method. In total, this modified method improved accuracy and efficiency of operations greatly, and would be realized a rapid and high-throughput determination of nitrate in field analysis of nitrate with low cost.

Exploring high-affinity binding properties of octamer peptides by principal component analysis of tetramer peptides.

PubMed

Kume, Akiko; Kawai, Shun; Kato, Ryuji; Iwata, Shinmei; Shimizu, Kazunori; Honda, Hiroyuki

2017-02-01

To investigate the binding properties of a peptide sequence, we conducted principal component analysis (PCA) of the physicochemical features of a tetramer peptide library comprised of 512 peptides, and the variables were reduced to two principal components. We selected IL-2 and IgG as model proteins and the binding affinity to these proteins was assayed using the 512 peptides mentioned above. PCA of binding affinity data showed that 16 and 18 variables were suitable for localizing IL-2 and IgG high-affinity binding peptides, respectively, into a restricted region of the PCA plot. We then investigated whether the binding affinity of octamer peptide libraries could be predicted using the identified region in the tetramer PCA. The results show that octamer high-affinity binding peptides were also concentrated in the tetramer high-affinity binding region of both IL-2 and IgG. The average fluorescence intensity of high-affinity binding peptides was 3.3- and 2.1-fold higher than that of low-affinity binding peptides for IL-2 and IgG, respectively. We conclude that PCA may be used to identify octamer peptides with high- or low-affinity binding properties from data from a tetramer peptide library. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

NITRATE POLLUTION IN SHALLOW GROUNDWATER OF A HARD ROCK REGION IN SOUTH CENTRAL INDIA

NASA Astrophysics Data System (ADS)

Brindha, K.; Rajesh, R.; Murugan, R.; Elango, L.

2009-12-01

Groundwater forms a major source of drinking water in most parts of the world. Due to the lack of piped drinking water supply, the population in rural areas depend on the groundwater resources for domestic purposes. Hence, the quality of groundwater in such regions needs to be monitored regularly. Presence of high concentration of nitrate in groundwater used for drinking is a major problem in many countries as it causes health related problems. Most often infants are affected by the intake of high nitrate in drinking water and food. The present study was carried out with the objective of assessing the nitrate concentration in groundwater and determining the causes for nitrate in groundwater in parts of Nalgonda district in India which is located at a distance of about 135 km towards ESE direction from Hyderabad. Nitrate concentration in groundwater of this area was analysed by collecting groundwater samples from forty six representative wells. Samples were collected once in two months from March 2008 to March 2009. A total of 244 groundwater samples were collected during the study. Soil samples were collected from fifteen locations during May 2009 and the denitrifying bacteria were isolated from the soil using spread plate method. The nitrate concentration in groundwater samples were analysed in the laboratory using Metrohm 861 advanced compact ion chromatograph using appropriate standards. The highest concentration of nitrate recorded during the sampling period was 879.65mg/l and the lowest concentration was below detection limit. The maximum permissible limit of nitrate for drinking water as per Bureau of Indian Standards is 45mg/l. About 13% of the groundwater samples collected from this study area possessed nitrate concentration beyond this limit. The nitrate concentration was high in the southeastern part of the study area. This implies that the nitrate concentration in groundwater tends to increase along the flow direction. Application of fertilizers is one of the sources for nitrate in groundwater. The recharge of rainwater through the indiscriminately dumped animal wastes also adds to nitrate in groundwater. As the population of denitrifying microbes (Agrobacterium sp.) in the topsoil increased, the nitrate concentration in groundwater decreased. The wells in the investigated region have been demarcated into safe and unsafe wells for consumption of water with respect to nitrate. The quality of groundwater in this region must be improved by denitrifying the groundwater before using it for consumption. Reduced dependence on nitrogen-rich fertilizers can also lower the influx of nitrates to a large extent. As the dumping of animal waste is also a reason behind high nitrate in groundwater, it would be better to use them as a biofertilizer. Due to the detrimental biological effects of nitrate, treatment and prevention methods must be considered to protect groundwater aquifers from nitrate leaching. Moreover, it is also important to educate the local population about keeping their surroundings clean, alternate use of the animal waste (as fuel) and to follow hygienic sanitation practices.

Respiration of Nitrate and Nitrite.

PubMed

Cole, Jeffrey A; Richardson, David J

2008-09-01

Nitrate reduction to ammonia via nitrite occurs widely as an anabolic process through which bacteria, archaea, and plants can assimilate nitrate into cellular biomass. Escherichia coli and related enteric bacteria can couple the eight-electron reduction of nitrate to ammonium to growth by coupling the nitrate and nitrite reductases involved to energy-conserving respiratory electron transport systems. In global terms, the respiratory reduction of nitrate to ammonium dominates nitrate and nitrite reduction in many electron-rich environments such as anoxic marine sediments and sulfide-rich thermal vents, the human gastrointestinal tract, and the bodies of warm-blooded animals. This review reviews the regulation and enzymology of this process in E. coli and, where relevant detail is available, also in Salmonella and draws comparisons with and implications for the process in other bacteria where it is pertinent to do so. Fatty acids may be present in high levels in many of the natural environments of E. coli and Salmonella in which oxygen is limited but nitrate is available to support respiration. In E. coli, nitrate reduction in the periplasm involves the products of two seven-gene operons, napFDAGHBC, encoding the periplasmic nitrate reductase, and nrfABCDEFG, encoding the periplasmic nitrite reductase. No bacterium has yet been shown to couple a periplasmic nitrate reductase solely to the cytoplasmic nitrite reductase NirB. The cytoplasmic pathway for nitrate reduction to ammonia is restricted almost exclusively to a few groups of facultative anaerobic bacteria that encounter high concentrations of environmental nitrate.

Effect of high oral doses of nitrate on salivary recirculation of nitrates and nitrites and on bacterial diversity in the saliva of young pigs.

PubMed

Trevisi, P; Casini, L; Nisi, I; Messori, S; Bosi, P

2011-04-01

Ingested nitrate is absorbed in the small intestine, recirculated into the saliva and reduced to nitrite by oral bacteria. In pigs receiving a moderate dietary addition of nitrate, the recirculation into the saliva is modest, so we aimed to assess the effect of higher nitrate doses to find out how the animal reacts to this new situation and to evaluate if a higher nitrate level could enhance the nitrate reduction process, improving the nitrite production Trial 1. Six piglets received 100 g of a commercial diet with 2.45% KNO(3) . In relation to baseline values, nitrate in blood serum and saliva increased 15 times, and declined after 6 h vs. 2 h. Salivary nitrite increased seven times after the addition and declined after 6 h vs. 2 h. Trial 2. Six piglets were fed a diet with or without 1.22% KNO(3) for 2 weeks. Salivary nitrate and nitrite increased with the addition of KNO3: nitrate increased from d0 to the end of the trial, nitrite increased 15 times after 1 week, but decreased after 2 weeks to 4.5-fold the control. After 2 weeks, nitrate reduced Shan diversity index of salivary microbiota. The present results indicate that the long exposure to high quantities of nitrates impairs the oral reduction of nitrate to nitrite and engenders a reduction of the mouth's microbiota diversity. © 2010 Blackwell Verlag GmbH.

Growth but Not Photosynthesis Response of a Host Plant to Infection by a Holoparasitic Plant Depends on Nitrogen Supply

PubMed Central

Shen, Hao; Xu, Shu-Jun; Hong, Lan; Wang, Zhang-Ming; Ye, Wan-Hui

2013-01-01

Parasitic plants can adversely influence the growth of their hosts by removing resources and by affecting photosynthesis. Such negative effects depend on resource availability. However, at varied resource levels, to what extent the negative effects on growth are attributed to the effects on photosynthesis has not been well elucidated. Here, we examined the influence of nitrogen supply on the growth and photosynthesis responses of the host plant Mikania micrantha to infection by the holoparasite Cuscuta campestris by focusing on the interaction of nitrogen and infection. Mikania micrantha plants fertilized at 0.2, 1 and 5 mM nitrate were grown with and without C. campestris infection. We observed that the infection significantly reduced M. micrantha growth at each nitrate fertilization and more severely at low than at high nitrate. Such alleviation at high nitrate was largely attributed to a stronger influence of infection on root biomass at low than at high nitrate fertilization. However, although C. campestris altered allometry and inhibited host photosynthesis, the magnitude of the effects was independent of nitrate fertilizations. The infection reduced light saturation point, net photosynthesis at saturating irradiances, apparent quantum yield, CO2 saturated rate of photosynthesis, carboxylation efficiency, the maximum carboxylation rate of Rubisco, and maximum light-saturated rate of electron transport, and increased light compensation point in host leaves similarly across nitrate levels, corresponding to a similar magnitude of negative effects of the parasite on host leaf soluble protein and Rubisco concentrations, photosynthetic nitrogen use efficiency and stomatal conductance across nitrate concentrations. Thus, the more severe inhibition in host growth at low than at high nitrate supplies cannot be attributed to a greater parasite-induced reduction in host photosynthesis, but the result of a higher proportion of host resources transferred to the parasite at low than at high nitrate levels. PMID:24116055

Vulnerability of shallow groundwater and drinking-water wells to nitrate in the United States

USGS Publications Warehouse

Nolan, Bernard T.; Hitt, Kerie J.

2006-01-01

Two nonlinear models were developed at the national scale to (1) predict contamination of shallow ground water (typically < 5 m deep) by nitrate from nonpoint sources and (2) to predict ambient nitrate concentration in deeper supplies used for drinking. The new models have several advantages over previous national-scale approaches. First, they predict nitrate concentration (rather than probability of occurrence), which can be directly compared with water-quality criteria. Second, the models share a mechanistic structure that segregates nitrogen (N) sources and physical factors that enhance or restrict nitrate transport and accumulation in ground water. Finally, data were spatially averaged to minimize small-scale variability so that the large-scale influences of N loading, climate, and aquifer characteristics could more readily be identified. Results indicate that areas with high N application, high water input, well-drained soils, fractured rocks or those with high effective porosity, and lack of attenuation processes have the highest predicted nitrate concentration. The shallow groundwater model (mean square error or MSE = 2.96) yielded a coefficient of determination (R2) of 0.801, indicating that much of the variation in nitrate concentration is explained by the model. Moderate to severe nitrate contamination is predicted to occur in the High Plains, northern Midwest, and selected other areas. The drinking-water model performed comparably (MSE = 2.00, R2 = 0.767) and predicts that the number of users on private wells and residing in moderately contaminated areas (>5 to ≤10 mg/L nitrate) decreases by 12% when simulation depth increases from 10 to 50 m.

Vulnerability of shallow groundwater and drinking-water wells to nitrate in the United States.

PubMed

Nolan, Bernard T; Hitt, Kerie J

2006-12-15

Two nonlinear models were developed at the national scale to (1) predict contamination of shallow ground water (typically < 5 m deep) by nitrate from nonpoint sources and (2) to predict ambient nitrate concentration in deeper supplies used for drinking. The new models have several advantages over previous national-scale approaches. First, they predict nitrate concentration (rather than probability of occurrence), which can be directly compared with water-quality criteria. Second, the models share a mechanistic structure that segregates nitrogen (N) sources and physical factors that enhance or restrict nitrate transport and accumulation in ground water. Finally, data were spatially averaged to minimize small-scale variability so that the large-scale influences of N loading, climate, and aquifer characteristics could more readily be identified. Results indicate that areas with high N application, high water input, well-drained soils, fractured rocks or those with high effective porosity, and lack of attenuation processes have the highest predicted nitrate concentration. The shallow groundwater model (mean square error or MSE = 2.96) yielded a coefficient of determination (R(2)) of 0.801, indicating that much of the variation in nitrate concentration is explained by the model. Moderate to severe nitrate contamination is predicted to occur in the High Plains, northern Midwest, and selected other areas. The drinking-water model performed comparably (MSE = 2.00, R(2) = 0.767) and predicts that the number of users on private wells and residing in moderately contaminated areas (>5 to < or =10 mg/L nitrate) decreases by 12% when simulation depth increases from 10 to 50 m.

Thin film superconductors and process for making same

DOEpatents

Nigrey, P.J.

1988-01-21

A process for the preparation of oxide superconductors from high-viscosity non-aqueous solution is described. Solutions of lanthanide nitrates, alkaline earth nitrates and copper nitrates in a 1:2:3 stoichiometric ratio, when added to ethylene glycol containing citric acid solutions, have been used to prepare highly viscous non-aqueous solutions of metal mixed nitrates-citrates. Thin films of these compositions are produced when a layer of the viscous solution is formed on a substrate and subjected to thermal decomposition.

Uptake of dissolved inorganic and organic nitrogen by the benthic toxic dinoflagellate Ostreopsis cf. ovata.

PubMed

Jauzein, Cécile; Couet, Douglas; Blasco, Thierry; Lemée, Rodolphe

2017-05-01

Environmental factors that shape dynamics of benthic toxic blooms are largely unknown. In particular, for the toxic dinoflagellate Ostreopsis cf. ovata, the importance of the availability of nutrients and the contribution of the inorganic and organic pools to growth need to be quantified in marine coastal environments. The present study aimed at characterizing N-uptake of dissolved inorganic and organic sources by O. cf. ovata cells, using the 15 N-labelling technique. Experiments were conducted taking into account potential interactions between nutrient uptake systems as well as variations with the diel cycle. Uptake abilities of O. cf. ovata were parameterized for ammonium (NH 4 + ), nitrate (NO 3 - ) and N-urea, from the estimation of kinetic and inhibition parameters. In the range of 0 to 10μmolNL -1 , kinetic curves showed a clear preference pattern following the ranking NH 4 + >NO 3 - >N-urea, where the preferential uptake of NH 4 + relative to NO 3 - was accentuated by an inhibitory effect of NH 4 + concentration on NO 3 - uptake capabilities. Conversely, under high nutrient concentrations, the preference for NH 4 + relative to NO 3 - was largely reduced, probably because of the existence of a low-affinity high capacity inducible NO 3 - uptake system. Ability to take up nutrients in darkness could not be defined as a competitive advantage for O. cf. ovata. Species competitiveness can also be defined from nutrient uptake kinetic parameters. A strong affinity for NH 4 + was observed for O. cf. ovata cells that may partly explain the success of this toxic species during the summer season in the Bay of Villefranche-sur-mer (France). Copyright © 2017 Elsevier B.V. All rights reserved.

Identification of Ftr1 and Zrt1 as iron and zinc micronutrient transceptors for activation of the PKA pathway in Saccharomyces cerevisiae

PubMed Central

Schothorst, Joep; Zeebroeck, Griet V.; Thevelein, Johan M.

2017-01-01

Multiple types of nutrient transceptors, membrane proteins that combine a transporter and receptor function, have now been established in a variety of organisms. However, so far all established transceptors utilize one of the macronutrients, glucose, amino acids, ammonium, nitrate, phosphate or sulfate, as substrate. This is also true for the Saccharomyces cerevisiae transceptors mediating activation of the PKA pathway upon re-addition of a macronutrient to glucose-repressed cells starved for that nutrient, re-establishing a fermentable growth medium. We now show that the yeast high-affinity iron transporter Ftr1 and high-affinity zinc transporter Zrt1 function as transceptors for the micronutrients iron and zinc. We show that replenishment of iron to iron-starved cells or zinc to zinc-starved cells triggers within 1-2 minutes a rapid surge in trehalase activity, a well-established PKA target. The activation with iron is dependent on Ftr1 and with zinc on Zrt1, and we show that it is independent of intracellular iron and zinc levels. Similar to the transceptors for macronutrients, Ftr1 and Zrt1 are strongly induced upon iron and zinc starvation, respectively, and they are rapidly downregulated by substrate-induced endocytosis. Our results suggest that transceptor-mediated signaling to the PKA pathway may occur in all cases where glucose-repressed yeast cells have been starved first for an essential nutrient, causing arrest of growth and low activity of the PKA pathway, and subsequently replenished with the lacking nutrient to re-establish a fermentable growth medium. The broadness of the phenomenon also makes it likely that nutrient transceptors use a common mechanism for signaling to the PKA pathway. PMID:28357393

Dietary nitrate supplementation in COPD: an acute, double-blind, randomized, placebo-controlled, crossover trial.

PubMed

Kerley, Conor P; Cahill, Kathleen; Bolger, Kenneth; McGowan, Aisling; Burke, Conor; Faul, John; Cormican, Liam

2015-01-30

The acute consumption of dietary nitrate has been shown to improve exercise capacity in athletes, healthy adults and subjects with peripheral vascular disease. Many COPD patients have reduced exercise capacity. We hypothesized that acute nitrate consumption might increase incremental shuttle walk test (ISWT) distance in COPD subjects. Eleven COPD subjects were randomly assigned to consume either a high nitrate or a matched, low nitrate beverage in a double-blind, randomized, placebo-controlled, crossover design. ISWT distance was measured both before and 3 h after the beverage and change was recorded. After a 7-day washout, ISWT distances were re-measured before and 3 h after the alternate beverage and changes were recorded. We observed an increase in ISWT distance after consuming the high nitrate juice (25 m) compared with a reduction after the low nitrate juice (14 m) (p < 0.01). This improvement in exercise capacity was associated with significant increases in serum nitrate (p < 0.000005) and nitrite (p < 0.01) levels and a significant lowering of resting blood pressure (<0.05). In patients with stable COPD, the acute consumption of dietary nitrate increased serum nitrate/nitrite levels and exercise capacity and was associated with a decrease in resting blood pressure. Nitrate consumption might alter exercise capacity in COPD patients. Copyright © 2014 Elsevier Inc. All rights reserved.

Hygroscopic behavior of atmospheric aerosols containing nitrate salts and water-soluble organic acids

NASA Astrophysics Data System (ADS)

Jing, Bo; Wang, Zhen; Tan, Fang; Guo, Yucong; Tong, Shengrui; Wang, Weigang; Zhang, Yunhong; Ge, Maofa

2018-04-01

While nitrate salts have critical impacts on environmental effects of atmospheric aerosols, the effects of coexisting species on hygroscopicity of nitrate salts remain uncertain. The hygroscopic behaviors of nitrate salt aerosols (NH4NO3, NaNO3, Ca(NO3)2) and their internal mixtures with water-soluble organic acids were determined using a hygroscopicity tandem differential mobility analyzer (HTDMA). The nitrate salt / organic acid mixed aerosols exhibit varying phase behavior and hygroscopic growth depending upon the type of components in the particles. Whereas pure nitrate salt particles show continuous water uptake with increasing relative humidity (RH), the deliquescence transition is still observed for ammonium nitrate particles internally mixed with organic acids such as oxalic acid and succinic acid with a high deliquescence point. The hygroscopicity of submicron aerosols containing sodium nitrate and an organic acid is also characterized by continuous growth, indicating that sodium nitrate tends to exist in a liquid-like state under dry conditions. It is observed that in contrast to the pure components, the water uptake is hindered at low and moderate RH for calcium nitrate particles containing malonic acid or phthalic acid, suggesting the potential effects of mass transfer limitation in highly viscous mixed systems. Our findings improve fundamental understanding of the phase behavior and water uptake of nitrate-salt-containing aerosols in the atmospheric environment.

Quantitative analysis of rat brain alpha 2-receptors discriminated by [3H]clonidine and [3H]rauwolscine.

PubMed

Asakura, M; Tsukamoto, T; Imafuku, J; Matsui, H; Ino, M; Hasegawa, K

1984-10-30

Quantitative analysis of direct ligand binding of both [3H]clonidine and [3H]rauwolscine to the rat cerebral cortex alpha 2-receptors indicates the existence of two affinity states of the same receptor populations. In the presence of Mn2+, the high affinity state of [3H]clonidine binding was increased, whereas the high affinity state of [3H]rauwolscine binding was reduced. By contrast, GTP in micromolar ranges caused a decrease of the agonist high affinity state and an increase of the antagonist high affinity state. The total receptor sites and the respective separate affinities for both radioligands were approximately equal to their control values under all conditions, indicating that Mn2+ and GTP modulate the proportion of the two affinity states of the receptor. These results can be incorporated into a two-step, ternary complex model involving a guanine nucleotide binding protein (N protein) for the agonist and antagonist interaction with the alpha 2-receptor. Furthermore, the effects of GTP on the interaction of both ligands with the two affinity states can be mimicked by EDTA. It is suggested that divalent cations induce the formation of the receptor-N protein binary complex showing high affinity for agonists and low affinity for antagonists.

Aquatic Nitrate Retention at River Network Scales Across Flow Conditions Determined Using Nested In Situ Sensors

NASA Astrophysics Data System (ADS)

Wollheim, W. M.; Mulukutla, G. K.; Cook, C.; Carey, R. O.

2017-11-01

Nonpoint pollution sources are strongly influenced by hydrology and are therefore sensitive to climate variability. Some pollutants entering aquatic ecosystems, e.g., nitrate, can be mitigated by in-stream processes during transport through river networks. Whole river network nitrate retention is difficult to quantify with observations. High frequency, in situ nitrate sensors, deployed in nested locations within a single watershed, can improve estimates of both nonpoint inputs and aquatic retention at river network scales. We deployed a nested sensor network and associated sampling in the urbanizing Oyster River watershed in coastal New Hampshire, USA, to quantify storm event-scale loading and retention at network scales. An end member analysis used the relative behavior of reactive nitrate and conservative chloride to infer river network fate of nitrate. In the headwater catchments, nitrate and chloride concentrations are both increasingly diluted with increasing storm size. At the mouth of the watershed, chloride is also diluted, but nitrate tended to increase. The end member analysis suggests that this pattern is the result of high retention during small storms (51-78%) that declines to zero during large storms. Although high frequency nitrate sensors did not alter estimates of fluxes over seasonal time periods compared to less frequent grab sampling, they provide the ability to estimate nitrate flux versus storm size at event scales that is critical for such analyses. Nested sensor networks can improve understanding of the controls of both loading and network scale retention, and therefore also improve management of nonpoint source pollution.

Molecular Hybridization of Potent and Selective γ-Hydroxybutyric Acid (GHB) Ligands: Design, Synthesis, Binding Studies, and Molecular Modeling of Novel 3-Hydroxycyclopent-1-enecarboxylic Acid (HOCPCA) and trans-γ-Hydroxycrotonic Acid (T-HCA) Analogs.

PubMed

Krall, Jacob; Jensen, Claus Hatt; Bavo, Francesco; Falk-Petersen, Christina Birkedahl; Haugaard, Anne Stæhr; Vogensen, Stine Byskov; Tian, Yongsong; Nittegaard-Nielsen, Mia; Sigurdardóttir, Sara Björk; Kehler, Jan; Kongstad, Kenneth Thermann; Gloriam, David E; Clausen, Rasmus Prætorius; Harpsøe, Kasper; Wellendorph, Petrine; Frølund, Bente

2017-11-09

γ-Hydroxybutyric acid (GHB) is a neuroactive substance with specific high-affinity binding sites. To facilitate target identification and ligand optimization, we herein report a comprehensive structure-affinity relationship study for novel ligands targeting these binding sites. A molecular hybridization strategy was used based on the conformationally restricted 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) and the linear GHB analog trans-4-hydroxycrotonic acid (T-HCA). In general, all structural modifications performed on HOCPCA led to reduced affinity. In contrast, introduction of diaromatic substituents into the 4-position of T-HCA led to high-affinity analogs (medium nanomolar K i ) for the GHB high-affinity binding sites as the most high-affinity analogs reported to date. The SAR data formed the basis for a three-dimensional pharmacophore model for GHB ligands, which identified molecular features important for high-affinity binding, with high predictive validity. These findings will be valuable in the further processes of both target characterization and ligand identification for the high-affinity GHB binding sites.

Evaluating sources and processing of nonpoint source nitrate in a small suburban watershed in China

NASA Astrophysics Data System (ADS)

Han, Li; Huang, Minsheng; Ma, Minghai; Wei, Jinbao; Hu, Wei; Chouhan, Seema

2018-04-01

Identifying nonpoint sources of nitrate has been a long-term challenge in mixed land-use watershed. In the present study, we combine dual nitrate isotope, runoff and stream water monitoring to elucidate the nonpoint nitrate sources across land use, and determine the relative importance of biogeochemical processes for nitrate export in a small suburban watershed, Longhongjian watershed, China. Our study suggested that NH4+ fertilizer, soil NH4+, litter fall and groundwater were the main nitrate sources in Longhongjian Stream. There were large changes in nitrate sources in response to season and land use. Runoff analysis illustrated that the tea plantation and forest areas contributed to a dominated proportion of the TN export. Spatial analysis illustrated that NO3- concentration was high in the tea plantation and forest areas, and δ15N-NO3 and δ18O-NO3 were enriched in the step ponds. Temporal analysis showed high NO3- level in spring, and nitrate isotopes were enriched in summer. Study as well showed that the step ponds played an important role in mitigating nitrate pollution. Nitrification and plant uptake were the significant biogeochemical processes contributing to the nitrogen transformation, and denitrification hardly occurred in the stream.

The origin of high-nitrate ground waters in the Australian arid zone

NASA Astrophysics Data System (ADS)

Barnes, C. J.; Jacobson, G.; Smith, G. D.

1992-08-01

Nitrate concentrations beyond the drinking-water limit of 10 mg1 -1 NO 3-N, are common in Australian arid-zone ground waters and are often associated with otherwise potable waters. In some aquifers nitrate-N concentrations of up to 80 mg1 -1 have been found, and this is a severe constraint on water supply development for small settlements. Water-bore data indicate a correlation of high-nitrate ground waters with shallow unconfined aquifers. Aguifer hydrochemistry indicats that these ground waters were emplaced by episodic Holocene recharge events in an otherwise arid climate regime. Nitrate has been flushed through the unsaturated zone which apparently lacks denitrification activity. The nitrate originates by near-surface biological fixation and contributing organisms include cyanobacteria in soil crusts and bacteria in termite mounds with the highest soil nitrate concentrations found in the outer skin of termite mounds. Bacteria associated with the termites appear to fix nitrogen, which eventually appears in an inorganic form, principally as ammonia. Nitrate is produced by bacterial oxidation of the ammonia, and is leached to the outside of the termite mound by capillary action. Diffuse recharge from extreme rainfall events then flushes this nitrate to the water table.

Nitrate Paradigm Does Not Hold Up for Sugarcane

PubMed Central

Robinson, Nicole; Brackin, Richard; Vinall, Kerry; Soper, Fiona; Holst, Jirko; Gamage, Harshi; Paungfoo-Lonhienne, Chanyarat; Rennenberg, Heinz; Lakshmanan, Prakash; Schmidt, Susanne

2011-01-01

Modern agriculture is based on the notion that nitrate is the main source of nitrogen (N) for crops, but nitrate is also the most mobile form of N and easily lost from soil. Efficient acquisition of nitrate by crops is therefore a prerequisite for avoiding off-site N pollution. Sugarcane is considered the most suitable tropical crop for biofuel production, but surprisingly high N fertilizer applications in main producer countries raise doubt about the sustainability of production and are at odds with a carbon-based crop. Examining reasons for the inefficient use of N fertilizer, we hypothesized that sugarcane resembles other giant tropical grasses which inhibit the production of nitrate in soil and differ from related grain crops with a confirmed ability to use nitrate. The results of our study support the hypothesis that N-replete sugarcane and ancestral species in the Andropogoneae supertribe strongly prefer ammonium over nitrate. Sugarcane differs from grain crops, sorghum and maize, which acquired both N sources equally well, while giant grass, Erianthus, displayed an intermediate ability to use nitrate. We conclude that discrimination against nitrate and a low capacity to store nitrate in shoots prevents commercial sugarcane varieties from taking advantage of the high nitrate concentrations in fertilized soils in the first three months of the growing season, leaving nitrate vulnerable to loss. Our study addresses a major caveat of sugarcane production and affords a strong basis for improvement through breeding cultivars with enhanced capacity to use nitrate as well as through agronomic measures that reduce nitrification in soil. PMID:21552564

A new class of organic nitrates: investigations on bioactivation, tolerance and cross-tolerance phenomena.

PubMed

Schuhmacher, S; Schulz, E; Oelze, M; König, A; Roegler, C; Lange, K; Sydow, L; Kawamoto, T; Wenzel, P; Münzel, T; Lehmann, J; Daiber, A

2009-09-01

The chronic use of organic nitrates is limited by serious side effects including oxidative stress, nitrate tolerance and/or endothelial dysfunction. The side effects and potency of nitroglycerine depend on mitochondrial aldehyde dehydrogenase (ALDH-2). We sought to determine whether this concept can be extended to a new class of organic nitrates with amino moieties (aminoalkyl nitrates). Vasodilator potency of the organic nitrates, in vitro tolerance and in vivo tolerance (after continuous infusion for 3 days) were assessed in wild-type and ALDH-2 knockout mice by isometric tension studies. Mitochondrial oxidative stress was analysed by L-012-dependent chemiluminescence and protein tyrosine nitration. Aminoethyl nitrate (AEN) showed an almost similar potency to glyceryl trinitrate (GTN), even though it is only a mononitrate. AEN-dependent vasodilatation was mediated by cGMP and nitric oxide. In contrast to triethanolamine trinitrate (TEAN) and GTN, AEN bioactivation did not depend on ALDH-2 and caused no in vitro tolerance. In vivo treatment with TEAN and GTN, but not with AEN, induced cross-tolerance to acetylcholine (ACh)-dependent and GTN-dependent relaxation. Although all nitrates tested induced tolerance to themselves, only TEAN and GTN significantly increased mitochondrial oxidative stress in vitro and in vivo. The present results demonstrate that not all high potency nitrates are bioactivated by ALDH-2 and that high potency of a given nitrate is not necessarily associated with induction of oxidative stress or nitrate tolerance. Obviously, there are distinct pathways for bioactivation of organic nitrates, which for AEN may involve xanthine oxidoreductase rather than P450 enzymes.

Risk of nitrate in groundwaters of the United States - A national perspective

USGS Publications Warehouse

Nolan, B.T.; Ruddy, B.C.; Hitt, K.J.; Helsel, D.R.

1997-01-01

Nitrate contamination of groundwater occurs in predictable patterns, based on findings of the U.S. Geological Survey's (USGS) National Water Quality Assessment (NAWQA) Program. The NAWQA Program was begun in 1991 to describe the quality of the Nation's water resources, using nationally consistent methods. Variables affecting nitrate concentration in groundwater were grouped as 'input' factors (population density end the amount of nitrogen contributed by fertilizer, manure, and atmospheric sources) and 'aquifer vulnerability' factors (soil drainage characteristic and the ratio of woodland acres to cropland acres in agricultural areas) and compiled in a national map that shows patterns of risk for nitrate contamination of groundwater. Areas with high nitrogen input, well-drained soils, and low woodland to cropland ratio have the highest potential for contamination of shallow groundwater by nitrate. Groundwater nitrate data collected through 1992 from wells less than 100 ft deep generally verified the risk patterns shown on the national map. Median nitrate concentration was 0.2 mg/L in wells representing the low-risk group, and the maximum contaminant level (MCL) was exceeded in 3% of the wells. In contrast, median nitrate concentration was 4.8 mg/L in wells representing the high-risk group, and the MCL was exceeded in 25% of the wells.Nitrate contamination of groundwater occurs in predictable patterns, based on findings of the U.S. Geological Survey's (USGS) National Water Quality Assessment (NAWQA) Program. The NAWQA Program was begun in 1991 to describe the quality of the Nation's water resources, using nationally consistent methods. Variables affecting nitrate concentration in groundwater were grouped as `input' factors (population density and the amount of nitrogen contributed by fertilizer, manure, and atmospheric sources) and `aquifer vulnerability' factors (soil drainage characteristic and the ratio of woodland acres to cropland acres in agricultural areas) and compiled in a national map that shows patterns of risk for nitrate contamination of groundwater. Areas with high nitrogen input, well-drained soils, and low woodland to cropland ratio have the highest potential for contamination of shallow groundwater by nitrate. Groundwater nitrate data collected through 1992 from wells less than 100 ft deep generally verified the risk patterns shown on the national map. Median nitrate concentration was 0.2 mg/L in wells representing the low-risk group, and the maximum contaminant level (MCL) was exceeded in 3% of the wells. In contrast, median nitrate concentration was 4.8 mg/L in wells representing the high-risk group, and the MCL was exceeded in 25% of the wells.

Ingested Nitrate and Breast Cancer in the Spanish Multicase-Control Study on Cancer (MCC-Spain).

PubMed

Espejo-Herrera, Nadia; Gracia-Lavedan, Esther; Pollan, Marina; Aragonés, Nuria; Boldo, Elena; Perez-Gomez, Beatriz; Altzibar, Jone M; Amiano, Pilar; Zabala, Ana Jiménez; Ardanaz, Eva; Guevara, Marcela; Molina, Antonio J; Barrio, Juan Pablo; Gómez-Acebo, Ines; Tardón, Adonina; Peiró, Rosana; Chirlaque, Maria Dolores; Palau, Margarita; Muñoz, Montse; Font-Ribera, Laia; Castaño-Vinyals, Gemma; Kogevinas, Manolis; Villanueva, Cristina M

2016-07-01

Ingested nitrate leads to endogenous formation of N-nitroso compounds that are breast carcinogens in animals, but human evidence is limited. We evaluated ingested nitrate as a risk factor for breast cancer (BC) in a multicase-control study. Hospital-based incident BC cases and population-based controls were recruited in eight Spanish regions in 2008-2013; participants provided residential and water consumption from 18 years of age and information on known BC risk factors. Long-term nitrate levels (1940-2010) were estimated and linked with residential histories and water consumption to calculate waterborne ingested nitrate (milligrams/day). Dietary ingested nitrate (milligrams/day) was calculated using food frequency questionnaires and published dietary nitrate contents. Interactions with endogenous nitrosation factors and other variables were evaluated. A total of 1,245 cases and 1,520 controls were included in the statistical analysis. Among the study regions, average ± SD waterborne ingested nitrate ranged from 2.9 ± 1.9 to 13.5 ± 7.5 mg/day, and dietary ingested nitrate ranged from 88.5 ± 48.7 to 154 ± 87.8 mg/day. Waterborne ingested nitrate was not associated with BC overall, but among postmenopausal women, those with both high nitrate (> 6 vs. < 2.6 mg/day) and high red meat intake (≥ 20 vs. < 20 g/day) were more likely to be cases than women with low nitrate and low red meat intake (adjusted odds ratio = 1.64; 95% confidence interval: 1.08, 2.49; overall interaction p-value = 0.17). No association was found with dietary nitrate. Waterborne ingested nitrate was associated with BC only among postmenopausal women with high red meat consumption. Dietary nitrate was not associated with BC regardless of the animal or vegetable source or of menopausal status. Espejo-Herrera N, Gracia-Lavedan E, Pollan M, Aragonés N, Boldo E, Perez-Gomez B, Altzibar JM, Amiano P, Zabala AJ, Ardanaz E, Guevara M, Molina AJ, Barrio JP, Gómez-Acebo I, Tardón A, Peiró R, Chirlaque MD, Palau M, Muñoz M, Font-Ribera L, Castaño-Vinyals G, Kogevinas M, Villanueva CM. 2016. Ingested nitrate and breast cancer in the Spanish Multicase-Control Study on Cancer (MCC-Spain). Environ Health Perspect 124:1042-1049; http://dx.doi.org/10.1289/ehp.1510334.

Effects of a Short-Term High-Nitrate Diet on Exercise Performance

PubMed Central

Porcelli, Simone; Pugliese, Lorenzo; Rejc, Enrico; Pavei, Gaspare; Bonato, Matteo; Montorsi, Michela; La Torre, Antonio; Rasica, Letizia; Marzorati, Mauro

2016-01-01

It has been reported that nitrate supplementation can improve exercise performance. Most of the studies have used either beetroot juice or sodium nitrate as a supplement; there is lack of data on the potential ergogenic benefits of an increased dietary nitrate intake from a diet based on fruits and vegetables. Our aim was to assess whether a high-nitrate diet increases nitric oxide bioavailability and to evaluate the effects of this nutritional intervention on exercise performance. Seven healthy male subjects participated in a randomized cross-over study. They were tested before and after 6 days of a high (HND) or control (CD) nitrate diet (~8.2 mmol∙day−1 or ~2.9 mmol∙day−1, respectively). Plasma nitrate and nitrite concentrations were significantly higher in HND (127 ± 64 µM and 350 ± 120 nM, respectively) compared to CD (23 ± 10 µM and 240 ± 100 nM, respectively). In HND (vs. CD) were observed: (a) a significant reduction of oxygen consumption during moderate-intensity constant work-rate cycling exercise (1.178 ± 0.141 vs. 1.269 ± 0.136 L·min−1); (b) a significantly higher total muscle work during fatiguing, intermittent sub-maximal isometric knee extension (357.3 ± 176.1 vs. 253.6 ± 149.0 Nm·s·kg−1); (c) an improved performance in Repeated Sprint Ability test. These findings suggest that a high-nitrate diet could be a feasible and effective strategy to improve exercise performance. PMID:27589795

Assessing the relationship between groundwater nitrate and animal feeding operations in Iowa (USA)

USGS Publications Warehouse

Zirkle, Keith W.; Nolan, Bernard T.; Jones, Rena R.; Weyer, Peter J.; Ward, Mary H.; Wheeler, David C.

2016-01-01

Nitrate-nitrogen is a common contaminant of drinking water in many agricultural areas of the United States of America (USA). Ingested nitrate from contaminated drinking water has been linked to an increased risk of several cancers, specific birth defects, and other diseases. In this research, we assessed the relationship between animal feeding operations (AFOs) and groundwater nitrate in private wells in Iowa. We characterized AFOs by swine and total animal units and type (open, confined, or mixed), and we evaluated the number and spatial intensities of AFOs in proximity to private wells. The types of AFO indicate the extent to which a facility is enclosed by a roof. Using linear regression models, we found significant positive associations between the total number of AFOs within 2 km of a well (p trend < 0.001), number of open AFOs within 5 km of a well (p trend < 0.001), and number of mixed AFOs within 30 km of a well (p trend < 0.001) and the log nitrate concentration. Additionally, we found significant increases in log nitrate in the top quartiles for AFO spatial intensity, open AFO spatial intensity, and mixed AFO spatial intensity compared to the bottom quartile (0.171 log(mg/L), 0.319 log(mg/L), and 0.541 log(mg/L), respectively; all p < 0.001). We also explored the spatial distribution of nitrate-nitrogen in drinking wells and found significant spatial clustering of high-nitrate wells (> 5 mg/L) compared with low-nitrate (≤ 5 mg/L) wells (p = 0.001). A generalized additive model for high-nitrate status identified statistically significant areas of risk for high levels of nitrate. Adjustment for some AFO predictor variables explained a portion of the elevated nitrate risk. These results support a relationship between animal feeding operations and groundwater nitrate concentrations and differences in nitrate loss from confined AFOs vs. open or mixed types.

The relationship between the nitrate concentration and hydrology of a small chalk spring; Israel

NASA Astrophysics Data System (ADS)

Burg, Avi; Heaton, Tim H. E.

1998-01-01

Discharge from a spring draining a small, perched, Cretaceous chalk aquifer in the Upper Galilee, Israel, was monitored over a period of two years. The water has elevated nitrate concentrations, with 15N/ 14N and chemical data suggesting that it is a mixture of low-nitrate and high-nitrate end-members; the latter derived from the sewage of a centuries-old village served by septic tanks. Hydrograph data allowed distinction between fissure flow during the period of winter rainfall, and matrix drainage during the dry summer months. These different flow types, however, did not have markedly different nitrate concentrations: a 50-fold increase in spring discharge due to fissure flow, compared with matrix drainage, was reflected in only a 35% decrease in nitrate concentrations. The relatively high nitrate concentrations in the fissure waters suggests that they have had close contact with, and are possibly displaced from the matrix. This should help to accelerate the decline in the spring's nitrate concentrations following the recent completion of the village's central sewage drainage system.

All-boron fullerene exhibits a strong affinity to inorganic anions

NASA Astrophysics Data System (ADS)

Colherinhas, Guilherme; Fileti, Eudes Eterno; Chaban, Vitaly V.

2017-03-01

Experimentally observed all-boron fullerene, B-80, inspires systematic investigation of its physical chemical properties and search for possible applications. We hereby report density functional theory calculations to characterize interactions of B-80 with the selected imidazolium room-temperature ionic liquids (RTILs), dimethylimidazolium nitrate and dimethylimidazolium hexafluorophosphate. Whereas the imidazolium cation exhibits a rather poor affinity to B-80, the inorganic anions form polar covalent bonds with the boron atom occupying a central position within a B-6 hexagon. Attachment of the RTIL ion pairs leads to a significant alteration of the electronic spectra, charge density distribution, valence and conduction molecular orbitals. The total binding energies keeping the RTIL@B80 complexes together range 200-250 kcal mol-1, being higher than the energies of many interactions in chemistry. The observed phenomenon predicts an excellent solubility of B-80 in the considered RTILs, but may also reveal a poor stability of B-80 in the polar media. Our results motivate further efforts in studying the behavior of the all-boron fullerene in polar environments.

Biological denitrification of high concentration nitrate waste

DOEpatents

Francis, Chester W.; Brinkley, Frank S.

1977-01-01

Biological denitrification of nitrate solutions at concentrations of greater than one kilogram nitrate per cubic meter is accomplished anaerobically in an upflow column having as a packing material a support for denitrifying bacteria.

Nitrate leaching in a winter wheat-summer maize rotation on a calcareous soil as affected by nitrogen and straw management.

PubMed

Huang, Tao; Ju, Xiaotang; Yang, Hao

2017-02-08

Nitrate leaching is one of the most important pathways of nitrogen (N) loss which leads to groundwater contamination or surface water eutrophication. Clarifying the rates, controlling factors and characteristics of nitrate leaching is the pre-requisite for proposing effective mitigation strategies. We investigated the effects of interactions among chemical N fertilizer, straw and manure applications on nitrogen leaching in an intensively managed calcareous Fluvo-aquic soil with winter wheat-summer maize cropping rotations on the North China Plain from October 2010 to September 2013 using ceramic suction cups and seepage water calculations based on a long-term field experiment. Annual nitrate leaching reached 38-60 kg N ha -1 from conventional N managements, but declined by 32-71% due to optimum N, compost manure or municipal waste treatments, respectively. Nitrate leaching concentrated in the summer maize season, and fewer leaching events with high amounts are the characteristics of nitrate leaching in this region. Overuse of chemical N fertilizers, high net mineralization and nitrification, together with predominance of rainfall in the summer season with light soil texture are the main controlling factors responsible for the high nitrate leaching loss in this soil-crop-climatic system.

Nitrate leaching in a winter wheat-summer maize rotation on a calcareous soil as affected by nitrogen and straw management

NASA Astrophysics Data System (ADS)

Huang, Tao; Ju, Xiaotang; Yang, Hao

2017-02-01

Nitrate leaching is one of the most important pathways of nitrogen (N) loss which leads to groundwater contamination or surface water eutrophication. Clarifying the rates, controlling factors and characteristics of nitrate leaching is the pre-requisite for proposing effective mitigation strategies. We investigated the effects of interactions among chemical N fertilizer, straw and manure applications on nitrogen leaching in an intensively managed calcareous Fluvo-aquic soil with winter wheat-summer maize cropping rotations on the North China Plain from October 2010 to September 2013 using ceramic suction cups and seepage water calculations based on a long-term field experiment. Annual nitrate leaching reached 38-60 kg N ha-1 from conventional N managements, but declined by 32-71% due to optimum N, compost manure or municipal waste treatments, respectively. Nitrate leaching concentrated in the summer maize season, and fewer leaching events with high amounts are the characteristics of nitrate leaching in this region. Overuse of chemical N fertilizers, high net mineralization and nitrification, together with predominance of rainfall in the summer season with light soil texture are the main controlling factors responsible for the high nitrate leaching loss in this soil-crop-climatic system.

Estimating discharge and non-point source nitrate loading to streams from three end-member pathways using high-frequency water quality and streamflow data

NASA Astrophysics Data System (ADS)

Miller, M. P.; Tesoriero, A. J.; Hood, K.; Terziotti, S.; Wolock, D.

2017-12-01

The myriad hydrologic and biogeochemical processes taking place in watersheds occurring across space and time are integrated and reflected in the quantity and quality of water in streams and rivers. Collection of high-frequency water quality data with sensors in surface waters provides new opportunities to disentangle these processes and quantify sources and transport of water and solutes in the coupled groundwater-surface water system. A new approach for separating the streamflow hydrograph into three components was developed and coupled with high-frequency specific conductance and nitrate data to estimate time-variable watershed-scale nitrate loading from three end-member pathways - dilute quickflow, concentrated quickflow, and slowflow groundwater - to two streams in central Wisconsin. Time-variable nitrate loads from the three pathways were estimated for periods of up to two years in a groundwater-dominated and a quickflow-dominated stream, using only streamflow and in-stream water quality data. The dilute and concentrated quickflow end-members were distinguished using high-frequency specific conductance data. Results indicate that dilute quickflow contributed less than 5% of the nitrate load at both sites, whereas 89±5% of the nitrate load at the groundwater-dominated stream was from slowflow groundwater, and 84±13% of the nitrate load at the quickflow-dominated stream was from concentrated quickflow. Concentrated quickflow nitrate concentrations varied seasonally at both sites, with peak concentrations in the winter that were 2-3 times greater than minimum concentrations during the growing season. Application of this approach provides an opportunity to assess stream vulnerability to non-point source nitrate loading and expected stream responses to current or changing conditions and practices in watersheds.

Identifying pathways and processes affecting nitrate and orthophosphate inputs to streams in agricultural watersheds

USGS Publications Warehouse

Tesoriero, A.J.; Duff, J.H.; Wolock, D.M.; Spahr, N.E.; Almendinger, J.E.

2009-01-01

Understanding nutrient pathways to streams will improve nutrient management strategies and estimates of the time lag between when changes in land use practices occur and when water quality effects that result from these changes are observed. Nitrate and orthophosphate (OP) concentrations in several environmental compartments were examined in watersheds having a range of base flow index (BFI) values across the continental United States to determine the dominant pathways for water and nutrient inputs to streams. Estimates of the proportion of stream nitrate that was derived from groundwater increased as BFI increased. Nitrate concentration gradients between groundwater and surface water further supported the groundwater source of nitrate in these high BFI streams. However, nitrate concentrations in stream-bed pore water in all settings were typically lower than stream or upland groundwater concentrations, suggesting that nitrate discharge to streams was not uniform through the bed. Rather, preferential pathways (e.g., springs, seeps) may allow high nitrate groundwater to bypass sites of high biogeochemical transformation. Rapid pathway compartments (e.g., overland flow, tile drains) had OP concentrations that were typically higher than in streams and were important OP conveyers in most of these watersheds. In contrast to nitrate, the proportion of stream OP that is derived from ground water did not systematically increase as BFI increased. While typically not the dominant source of OP, groundwater discharge was an important pathway of OP transport to streams when BFI values were very high and when geochemical conditions favored OP mobility in groundwater. Copyright ?? 2009 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

Effects of heat shock, stannous chloride, and gallium nitrate on the rat inflammatory response

PubMed Central

House, S.D.; Guidon, P.T.; Perdrizet, G.A.; Rewinski, M.; Kyriakos, R.; Bockman, R.S.; Mistry, T.; Gallagher, P.A.; Hightower, L.E.

2001-01-01

Heat and a variety of other stressors cause mammalian cells and tissues to acquire cytoprotection. This transient state of altered cellular physiology is nonproliferative and antiapoptotic. In this study, male Wistar rats were stress conditioned with either stannous chloride or gallium nitrate, which have immunosuppressive effects in vivo and in vitro, or heat shock, the most intensively studied inducer of cytoprotection. The early stages of inflammation in response to topical suffusion of mesentery tissue with formyl-methionyl-leucyl-phenylalanine (FMLP) were monitored using intravital microscopy. Microvascular hemodynamics (venular diameter, red blood cell velocity [Vrbc], white blood cell [WBC] flux, and leukocyte-endothelial adhesion [LEA]) were used as indicators of inflammation, and tissue levels of inducible Hsp70, determined using immunoblot assays, provided a marker of cytoprotection. None of the experimental treatments blocked decreases in WBC flux during FMLP suffusion, an indicator of increased low-affinity interactions between leukocytes and vascular endothelium known as rolling adhesion. During FMLP suffusion LEA, an indicator of firm attachment between leukocytes and vascular endothelial cells increased in placebo and gallium nitrate-treated animals but not in heat- and stannous chloride–treated animals, an anti-inflammatory effect. Hsp70 was not detected in aortic tissue from placebo and gallium nitrate–treated animals, indicating that Hsp70-dependent cytoprotection was not present. In contrast, Hsp70 was detected in aortic tissues from heat- and stannous chloride–treated animals, indicating that these tissues were in a cytoprotected state that was also an anti-inflammatory state. PMID:11599578

Post-translational regulation of nitrogen transporters in plants and microorganisms.

PubMed

Jacquot, Aurore; Li, Zhi; Gojon, Alain; Schulze, Waltraud; Lejay, Laurence

2017-05-01

For microorganisms and plants, nitrate and ammonium are the main nitrogen sources and they are also important signaling molecules controlling several aspects of metabolism and development. Over the past decade, numerous studies revealed that nitrogen transporters are strongly regulated at the transcriptional level. However, more and more reports are now showing that nitrate and ammonium transporters are also subjected to post-translational regulations in response to nitrogen availability. Phosphorylation is so far the most well studied post-translational modification for these transporters and it affects both the regulation of nitrogen uptake and nitrogen sensing. For example, in Arabidopsis thaliana, phosphorylation was shown to activate the sensing function of the root nitrate transporter NRT1.1 and to switch the transport affinity. Also, for ammonium transporters, a phosphorylation-dependent activation/inactivation mechanism was elucidated in recent years in both plants and microorganisms. However, despite the fact that these regulatory mechanisms are starting to be thoroughly described, the signaling pathways involved and their action on nitrogen transporters remain largely unknown. In this review, we highlight the inorganic nitrogen transporters regulated at the post-translational level and we compare the known mechanisms in plants and microorganisms. We then discuss how these mechanisms could contribute to the regulation of nitrogen uptake and/or nitrogen sensing. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Engineering an antibody with picomolar affinity to DOTA chelates of multiple radionuclides for pretargeted radioimmunotherapy and imaging

PubMed Central

Orcutt, Kelly Davis; Slusarczyk, Adrian L; Cieslewicz, Maryelise; Ruiz-Yi, Benjamin; Bhushan, Kumar R; Frangioni, John V; Wittrup, K Dane

2014-01-01

Introduction In pretargeted radioimmunotherapy (PRIT), a bifunctional antibody is administered and allowed to pre-localize to tumor cells. Subsequently, a chelated radionuclide is administered and captured by cell-bound antibody while unbound hapten clears rapidly from the body. We aim to engineer high-affinity binders to DOTA chelates for use in PRIT applications. Methods We mathematically modeled antibody and hapten pharmacokinetics to analyze hapten tumor retention as a function of hapten binding affinity. Motivated by model predictions, we used directed evolution and yeast surface display to affinity mature the 2D12.5 antibody to 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), reformatted as a single chain variable fragment (scFv). Results Modeling predicts that for high antigen density and saturating bsAb dose, a hapten binding affinity of 100 picomolar (pM) is needed for near-maximal hapten retention. We affinity matured 2D12.5 with an initial binding constant of about 10 nanomolar (nM) to DOTA-yttrium chelates. Affinity maturation resulted in a 1000-fold affinity improvement to biotinylated DOTA-yttrium, yielding an 8.2 ± 1.9 picomolar binder. The high-affinity scFv binds DOTA complexes of lutetium and gadolinium with similar picomolar affinity and indium chelates with low nanomolar affinity. When engineered into a bispecific antibody construct targeting carcinoembryonic antigen (CEA), pretargeted high-affinity scFv results in significantly higher tumor retention of a 111In-DOTA hapten compared to pretargeted wild-type scFv in a xenograft mouse model. Conclusions We have engineered a versatile, high-affinity DOTA-chelate-binding scFv. We anticipate it will prove useful in developing pretargeted imaging and therapy protocols to exploit the potential of a variety of radiometals. PMID:21315278

Characterization of the Heme Pocket Structure and Ligand Binding Kinetics of Non-symbiotic Hemoglobins from the Model Legume Lotus japonicus.

PubMed

Calvo-Begueria, Laura; Cuypers, Bert; Van Doorslaer, Sabine; Abbruzzetti, Stefania; Bruno, Stefano; Berghmans, Herald; Dewilde, Sylvia; Ramos, Javier; Viappiani, Cristiano; Becana, Manuel

2017-01-01

Plant hemoglobins (Hbs) are found in nodules of legumes and actinorhizal plants but also in non-symbiotic organs of monocots and dicots. Non-symbiotic Hbs (nsHbs) have been classified into two phylogenetic groups. Class 1 nsHbs show an extremely high O 2 affinity and are induced by hypoxia and nitric oxide (NO), whereas class 2 nsHbs have moderate O 2 affinity and are induced by cold and cytokinins. The functions of nsHbs are still unclear, but some of them rely on the capacity of hemes to bind diatomic ligands and catalyze the NO dioxygenase (NOD) reaction (oxyferrous Hb + NO → ferric Hb + nitrate). Moreover, NO may nitrosylate Cys residues of proteins. It is therefore important to determine the ligand binding properties of the hemes and the role of Cys residues. Here, we have addressed these issues with the two class 1 nsHbs (LjGlb1-1 and LjGlb1-2) and the single class 2 nsHb (LjGlb2) of Lotus japonicus , which is a model legume used to facilitate the transfer of genetic and biochemical information into crops. We have employed carbon monoxide (CO) as a model ligand and resonance Raman, laser flash photolysis, and stopped-flow spectroscopies to unveil major differences in the heme environments and ligand binding kinetics of the three proteins, which suggest non-redundant functions. In the deoxyferrous state, LjGlb1-1 is partially hexacoordinate, whereas LjGlb1-2 shows complete hexacoordination (behaving like class 2 nsHbs) and LjGlb2 is mostly pentacoordinate (unlike other class 2 nsHbs). LjGlb1-1 binds CO very strongly by stabilizing it through hydrogen bonding, but LjGlb1-2 and LjGlb2 show lower CO stabilization. The changes in CO stabilization would explain the different affinities of the three proteins for gaseous ligands. These affinities are determined by the dissociation rates and follow the order LjGlb1-1 > LjGlb1-2 > LjGlb2. Mutations LjGlb1-1 C78S and LjGlb1-2 C79S caused important alterations in protein dynamics and stability, indicating a structural role of those Cys residues, whereas mutation LjGlb1-1 C8S had a smaller effect. The three proteins and their mutant derivatives exhibited similarly high rates of NO consumption, which were due to NOD activity of the hemes and not to nitrosylation of Cys residues.

Characterization of the Heme Pocket Structure and Ligand Binding Kinetics of Non-symbiotic Hemoglobins from the Model Legume Lotus japonicus

PubMed Central

Calvo-Begueria, Laura; Cuypers, Bert; Van Doorslaer, Sabine; Abbruzzetti, Stefania; Bruno, Stefano; Berghmans, Herald; Dewilde, Sylvia; Ramos, Javier; Viappiani, Cristiano; Becana, Manuel

2017-01-01

Plant hemoglobins (Hbs) are found in nodules of legumes and actinorhizal plants but also in non-symbiotic organs of monocots and dicots. Non-symbiotic Hbs (nsHbs) have been classified into two phylogenetic groups. Class 1 nsHbs show an extremely high O2 affinity and are induced by hypoxia and nitric oxide (NO), whereas class 2 nsHbs have moderate O2 affinity and are induced by cold and cytokinins. The functions of nsHbs are still unclear, but some of them rely on the capacity of hemes to bind diatomic ligands and catalyze the NO dioxygenase (NOD) reaction (oxyferrous Hb + NO → ferric Hb + nitrate). Moreover, NO may nitrosylate Cys residues of proteins. It is therefore important to determine the ligand binding properties of the hemes and the role of Cys residues. Here, we have addressed these issues with the two class 1 nsHbs (LjGlb1-1 and LjGlb1-2) and the single class 2 nsHb (LjGlb2) of Lotus japonicus, which is a model legume used to facilitate the transfer of genetic and biochemical information into crops. We have employed carbon monoxide (CO) as a model ligand and resonance Raman, laser flash photolysis, and stopped-flow spectroscopies to unveil major differences in the heme environments and ligand binding kinetics of the three proteins, which suggest non-redundant functions. In the deoxyferrous state, LjGlb1-1 is partially hexacoordinate, whereas LjGlb1-2 shows complete hexacoordination (behaving like class 2 nsHbs) and LjGlb2 is mostly pentacoordinate (unlike other class 2 nsHbs). LjGlb1-1 binds CO very strongly by stabilizing it through hydrogen bonding, but LjGlb1-2 and LjGlb2 show lower CO stabilization. The changes in CO stabilization would explain the different affinities of the three proteins for gaseous ligands. These affinities are determined by the dissociation rates and follow the order LjGlb1-1 > LjGlb1-2 > LjGlb2. Mutations LjGlb1-1 C78S and LjGlb1-2 C79S caused important alterations in protein dynamics and stability, indicating a structural role of those Cys residues, whereas mutation LjGlb1-1 C8S had a smaller effect. The three proteins and their mutant derivatives exhibited similarly high rates of NO consumption, which were due to NOD activity of the hemes and not to nitrosylation of Cys residues. PMID:28421084

Dietary nitrate as modulator of physical performance and cardiovascular health.

PubMed

Kerley, Conor P

2017-11-01

Early interventional trials reported improvements in cardiac and exercise outcomes with inorganic nitrate ingestion. The current review aims to provide a brief update of recent evidence regarding ergogenic and cardiovascular effects of dietary nitrate and practical recommendations. Recent evidence has been inconsistent and questions remain regarding effective dose, duration, and source of nitrate and cohorts likely to benefit. Dietary nitrate may be most relevant to those with vascular/metabolic impairments, those engaging in short-term, intense exercise, deconditioned individuals, and those with a low dietary nitrate intake. The evidence for cardiovascular/exercise benefit is plausible but inconsistent. However, dietary nitrate, in contrast to pharmacological nitrate, has a high benefit-risk ratio. Although nitrate supplementation has grown in popularity, it is suggested that increased green vegetables consumption may provide similar/superior benefits to nitrate supplementation in a cheaper, safer, and potentially tastier context.

The preservation of long-range transported nitrate in snow at Summit, Greenland (Invited)

NASA Astrophysics Data System (ADS)

Hastings, M. G.

2013-12-01

Nitrate is one of the major anions found in polar and alpine snow, both today and in the past. Deposition of nitrate to snow surfaces results from reactions of nitrogen oxides (NOx) with oxidants in the atmosphere, resulting in the production of HNO3 that is incorporated into precipitation or reacts on the surface of particles. Several factors motivate studying nitrate concentration in ice cores including reconstructing past levels of NOx, tropospheric oxidant concentrations and natural variability in NOx sources. The link between the atmospheric concentration of NOx and nitrate concentration in ice core records is problematic because post-depositional processing, such as photolysis and evaporation, can impact the concentration of nitrate in snow. Recent work has shown that the isotopic ratios of nitrate (15N/14N, 18O/16O, 17O/16O) can be a powerful tool for tracing post-depositional loss of nitrate from surface snow. The isotopic composition of nitrate has been shown to contain information about the source of the nitrate (i.e, NOx sources) and the oxidation processes that convert NOx to nitrate in the atmosphere prior to deposition. Results from a number of studies at Summit, Greenland reveal limited loss of nitrate from surface snow during highly photoactive periods, and the oxygen isotopic signatures in snow nitrate appear to be representative of atmospheric deposition of nitrate from outside of Summit. Higher than expected oxygen isotope ratios (18O/16O, 17O/16O) found in Summit summertime nitrate were expected to be dependent upon local photochemistry in which nitrate in the snow is photolyzed to NOx that is then oxidized above the snow by BrO to reform nitrate (i.e., BrONO2). However, the oxygen isotopic composition of nitrate collected at high time resolution in surface snow does not show any link to local gas phase concentrations of a number of species, including BrO. Furthermore, the combination of nitrogen and oxygen isotope data reveals interesting insights into the contributions of nitrate sources to Summit. There are several important implications of this work including that nitrate at Summit appears to be largely preserved in surface snow during photoactive periods, and that nitrate in snow at Summit also appears to be representative of long-range transported nitrate/NOx. The surface snow work is further substantiated by relationships found between and among seasonally-resolved ice core measurements of the isotopic composition of nitrate, nitrate concentration and a suite of chemical and elemental tracers. The seasonality observed in 15N/14N ratios in an ice core representing accumulation since 1760 C.E. cannot be explained by diffusion or other processes occurring in the firn over time. A marked negative trend in 15N/14N since industrialization, parallels a nearly three-fold increase in nitrate concentration as well as pronounced increases in tracers such as excess lead and non-sea-salt sulfur. This, along with independent estimates of oil burning and transport studies, indicate that North American oil combustion is the primary driver of the modern negative trend in 15N/14N of nitrate. The high, positive 15N/14N ratios found in pre-industrial ice link to biomass burning based upon concentrations of black carbon and ammonium.

Nitrate and amino acid availability affects glycine betaine and mycosporine-2-glycine in response to changes of salinity in a halotolerant cyanobacterium Aphanothece halophytica.

PubMed

Waditee-Sirisattha, Rungaroon; Kageyama, Hakuto; Fukaya, Minoru; Rai, Vandna; Takabe, Teruhiro

2015-12-01

A halotolerant cyanobacterium Aphanothece halophytica thrives in extreme salinity with accumulation of a potent osmoprotectant glycine betaine. Recently, this cyanobacterium was shown to accumulate sunscreen molecule mycosporine-2-glycine significantly at high salinity. In this study, we investigated effects of nitrate and amino acid provision on the accumulation of glycine betaine and mycosporine-2-glycine. With elevated nitrate concentrations at high salinity, intracellular levels of both metabolites were enhanced. Six-fold high nitrate concentration increased the relative amounts of glycine betaine and mycosporine-2-glycine to be 1.5 and 2.0 folds compared with control condition : Increased levels were time- and dose-dependent manner. Exogenous supply of glycine/serine at high salinity resulted in the similar trends as observed in excess nitrate experiment. Intracellular level of glycine betaine increased ∼1.6 folds with glycine/serine supplementation. These supplementations also caused the increased level of mycosporine-2-glycine, namely 1.4 and 2 folds by glycine and serine, respectively. The transcription of glycine betaine and mycosporine-2-glycine biosynthetic genes was strongly induced under high-nitrate-salt condition. These results suggest the dependence of glycine betaine and mycosporine-2-glycine productions on substrate availability, and the effect of nitrate was possibly associated with stimulation of osmoprotectant increment in this extremophile. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Effects of Atmospheric Nitrate on an Upland Stream of the Northeastern USA

NASA Astrophysics Data System (ADS)

Sebestyen, S. D.; Shanley, J. B.; Boyer, E. W.; Kendall, C.

2009-05-01

Excess nitrogen cascades through terrestrial biogeochemical cycles and affects stream nitrate concentrations in upland forests where atmospheric deposition is an important source of anthropogenic nitrogen. We will discuss approaches including high-frequency sampling, isotopic tracers, and end-member mixing analysis that can be used to decipher the sources, transformations, and hydrological processes that affect nitrate transport through forested upland catchments to streams. We present results of studies at the Sleepers River Research Watershed in Vermont, USA, a site where we have intensively measured stream nitrate concentrations during baseflow and stormflow. Stream nitrate concentrations are typically low and nearly 75% of annual inorganic N inputs from atmospheric deposition are retained within the catchment. However, high concentrations and stream loadings of nitrate occur during storm events due to source variation and hydrological flushing of nitrate from catchment soils. Using isotopic tracers and end-member mixing analysis, we have quantified source inputs of unprocessed atmospheric nitrate and show that this stream is directly affected by nitrogen pollution. Using a long-term record of stream hydrochemistry and our findings on event- scale nitrate flushing dynamics, we then explore how stream nitrate loading may respond to anthropogenic climate forcing during the next century. Results suggest that stream runoff and nitrate loadings will change during future emission scenarios (i.e. longer growing seasons and higher winter precipitation rates). Understanding the timing and magnitude of hydrological and hydrochemical responses is important because climate change effects on catchment hydrology may alter how nitrate is retained, produced, and hydrologically flushed in headwater ecosystems with implications for aquatic metabolism, nutrient export from catchments, and downstream eutrophication.

A new class of organic nitrates: investigations on bioactivation, tolerance and cross-tolerance phenomena

PubMed Central

Schuhmacher, S; Schulz, E; Oelze, M; König, A; Roegler, C; Lange, K; Sydow, L; Kawamoto, T; Wenzel, P; Münzel, T; Lehmann, J; Daiber, A

2009-01-01

Background and purpose: The chronic use of organic nitrates is limited by serious side effects including oxidative stress, nitrate tolerance and/or endothelial dysfunction. The side effects and potency of nitroglycerine depend on mitochondrial aldehyde dehydrogenase (ALDH-2). We sought to determine whether this concept can be extended to a new class of organic nitrates with amino moieties (aminoalkyl nitrates). Experimental approach: Vasodilator potency of the organic nitrates, in vitro tolerance and in vivo tolerance (after continuous infusion for 3 days) were assessed in wild-type and ALDH-2 knockout mice by isometric tension studies. Mitochondrial oxidative stress was analysed by L-012-dependent chemiluminescence and protein tyrosine nitration. Key results: Aminoethyl nitrate (AEN) showed an almost similar potency to glyceryl trinitrate (GTN), even though it is only a mononitrate. AEN-dependent vasodilatation was mediated by cGMP and nitric oxide. In contrast to triethanolamine trinitrate (TEAN) and GTN, AEN bioactivation did not depend on ALDH-2 and caused no in vitro tolerance. In vivo treatment with TEAN and GTN, but not with AEN, induced cross-tolerance to acetylcholine (ACh)-dependent and GTN-dependent relaxation. Although all nitrates tested induced tolerance to themselves, only TEAN and GTN significantly increased mitochondrial oxidative stress in vitro and in vivo. Conclusions and implications: The present results demonstrate that not all high potency nitrates are bioactivated by ALDH-2 and that high potency of a given nitrate is not necessarily associated with induction of oxidative stress or nitrate tolerance. Obviously, there are distinct pathways for bioactivation of organic nitrates, which for AEN may involve xanthine oxidoreductase rather than P450 enzymes. PMID:19563531

Evaluation on the Nanoscale Zero Valent Iron Based Microbial Denitrification for Nitrate Removal from Groundwater

NASA Astrophysics Data System (ADS)

Peng, Lai; Liu, Yiwen; Gao, Shu-Hong; Chen, Xueming; Xin, Pei; Dai, Xiaohu; Ni, Bing-Jie

2015-07-01

Nanoscale zero valent iron (NZVI) based microbial denitrification has been demonstrated to be a promising technology for nitrate removal from groundwater. In this work, a mathematical model is developed to evaluate the performance of this new technology and to provide insights into the chemical and microbial interactions in the system in terms of nitrate reduction, ammonium accumulation and hydrogen turnover. The developed model integrates NZVI-based abiotic reduction of nitrate, NZVI corrosion for hydrogen production and hydrogen-based microbial denitrification and satisfactorily describes all of the nitrate and ammonium dynamics from two systems with highly different conditions. The high NZVI corrosion rate revealed by the model indicates the high reaction rate of NZVI with water due to their large specific surface area and high surface reactivity, leading to an effective microbial nitrate reduction by utilizing the produced hydrogen. The simulation results further suggest a NZVI dosing strategy (3-6 mmol/L in temperature range of 30-40 °C, 6-10 mmol/L in temperature range of 15-30 °C and 10-14 mmol/L in temperature range of 5-15 °C) during groundwater remediation to make sure a low ammonium yield and a high nitrogen removal efficiency.

Evaluation on the Nanoscale Zero Valent Iron Based Microbial Denitrification for Nitrate Removal from Groundwater

PubMed Central

Peng, Lai; Liu, Yiwen; Gao, Shu-Hong; Chen, Xueming; Xin, Pei; Dai, Xiaohu; Ni, Bing-Jie

2015-01-01

Nanoscale zero valent iron (NZVI) based microbial denitrification has been demonstrated to be a promising technology for nitrate removal from groundwater. In this work, a mathematical model is developed to evaluate the performance of this new technology and to provide insights into the chemical and microbial interactions in the system in terms of nitrate reduction, ammonium accumulation and hydrogen turnover. The developed model integrates NZVI-based abiotic reduction of nitrate, NZVI corrosion for hydrogen production and hydrogen-based microbial denitrification and satisfactorily describes all of the nitrate and ammonium dynamics from two systems with highly different conditions. The high NZVI corrosion rate revealed by the model indicates the high reaction rate of NZVI with water due to their large specific surface area and high surface reactivity, leading to an effective microbial nitrate reduction by utilizing the produced hydrogen. The simulation results further suggest a NZVI dosing strategy (3–6 mmol/L in temperature range of 30–40 °C, 6–10 mmol/L in temperature range of 15–30 °C and 10–14 mmol/L in temperature range of 5–15 °C) during groundwater remediation to make sure a low ammonium yield and a high nitrogen removal efficiency. PMID:26199053

Quantifying the fate of agricultural nitrogen in an unconfined aquifer: Stream-based observations at three measurement scales

NASA Astrophysics Data System (ADS)

Gilmore, Troy E.; Genereux, David P.; Solomon, D. Kip; Solder, John E.; Kimball, Briant A.; Mitasova, Helena; Birgand, François

2016-03-01

We compared three stream-based sampling methods to study the fate of nitrate in groundwater in a coastal plain watershed: point measurements beneath the streambed, seepage blankets (novel seepage-meter design), and reach mass-balance. The methods gave similar mean groundwater seepage rates into the stream (0.3-0.6 m/d) during two 3-4 day field campaigns despite an order of magnitude difference in stream discharge between the campaigns. At low flow, estimates of flow-weighted mean nitrate concentrations in groundwater discharge ([NO3-]FWM) and nitrate flux from groundwater to the stream decreased with increasing degree of channel influence and measurement scale, i.e., [NO3-]FWM was 654, 561, and 451 µM for point, blanket, and reach mass-balance sampling, respectively. At high flow the trend was reversed, likely because reach mass-balance captured inputs from shallow transient high-nitrate flow paths while point and blanket measurements did not. Point sampling may be better suited to estimating aquifer discharge of nitrate, while reach mass-balance reflects full nitrate inputs into the channel (which at high flow may be more than aquifer discharge due to transient flow paths, and at low flow may be less than aquifer discharge due to channel-based nitrate removal). Modeling dissolved N2 from streambed samples suggested (1) about half of groundwater nitrate was denitrified prior to discharge from the aquifer, and (2) both extent of denitrification and initial nitrate concentration in groundwater (700-1300 µM) were related to land use, suggesting these forms of streambed sampling for groundwater can reveal watershed spatial relations relevant to nitrate contamination and fate in the aquifer.

Relation of nitrate concentrations to baseflow in the Raccoon River, Iowa

USGS Publications Warehouse

Schilling, K.E.; Lutz, D.S.

2004-01-01

Excessive nitrate-nitrogen (nitrate) export from the Raccoon River in west central Iowa is an environmental concern to downstream receptors. The 1972 to 2000 record of daily streamflow and the results from 981 nitrate measurements were examined to describe the relation of nitrate to streamflow in the Raccoon River. No long term trends in streamflow and nitrate concentrations were noted in the 28-year record. Strong seasonal patterns were evident in nitrate concentrations, with higher concentrations occurring in spring and fall. Nitrate concentrations were linearly related to streamflow at daily, monthly, seasonal, and annual time scales. At all time scales evaluated, the relation was improved when baseflow was used as the discharge variable instead of total streamflow. Nitrate concentrations were found to be highly stratified according to flow, but there was little relation of nitrate to streamflow within each flow range. Simple linear regression models developed to predict monthly mean nitrate concentrations explained as much as 76 percent of the variability in the monthly nitrate concentration data for 2001. Extrapolation of current nitrate baseflow relations to historical conditions in the Raccoon River revealed that increasing baseflow over the 20th century could account for a measurable increase in nitrate concentrations.

Point mutation increases a form of the NK1 receptor with high affinity for neurokinin A and B and septide

PubMed Central

Ciucci, Alessandra; Palma, Carla; Manzini, Stefano; Werge, Thomas M

1998-01-01

The binding modalities of substance P and neurokinin A on the wild type and Gly166 to-Cys mutant NK1 receptors expressed on CHO cells were investigated in homologous and heterologous binding experiments using both radiolabelled substance P and neurokinin A.On the wild type NK1 receptor NKA displaces radiolabelled substance P with very low apparent affinity, despite its high-affinity binding constant (determined in homologous binding experiments). The Gly166 to-Cys substitution in the NK1 tachykinin receptor greatly enhances the apparent affinity of neurokinin A in competition for radiolabelled substance P, but it does not change the binding constant of neurokinin A. The mutation, thereby, eliminates the discrepancy between the low apparent affinity and the high binding constant of neurokinin A.On the wild type receptor the binding capacity of neurokinin A is significantly smaller than that of substance P. In contrast, the two tachykinins bind to approximately the same number of sites on the mutant receptor.Simultaneous mass action law analysis of binding data in which multiple radioligands were employed in parallel demonstrated that a one-site model was unable to accommodate all the experimental data, whereas a two-site model provided a dramatically better description.These two receptor-sites display equally high affinity for substance P, while neurokinin A strongly discriminates between a high and a low affinity component. The binding affinities of neurokinin A are not affected by the mutation, which instead specifically alters the distribution between receptor sites in favour of a high affinity neurokinin A binding form.The low apparent affinity and binding capacity of neurokinin A on the wild type receptor results from neurokinin A binding with high affinity only to a fraction of the sites labelled by substance P. The mutation increases the proportion of this site, and consequently enhances the apparent affinity and binding capacity of neurokinin A.The binding modalities of septide-like ligands (i.e. neurokinin B, SP(6-11), SP-methyl ester) are affected similarly to neurokinin A and are better resolved into two sites. The mutation leaves the affinity of these ligands for the two receptor forms unchanged, but increases the fraction of high-affinity sites. On the other hand, the binding of non-peptide and peptide antagonists (SR140.333 and FK888) behaved similarly to substance P with a single high affinity site that is unaffected by the mutation.These findings may suggest that the NK1 receptor exists in two different forms with similar affinity for substance P and NK1 antagonists, but with a high and a low affinity for neurokinin A and septide-like ligands. Hence, the Gly166 in the NK1 receptor would seem to control the distribution between a pan-reactive form and a substance P-selective form of the receptor. PMID:9786514

Biotransformation of RDX and HMX by Anaerobic Granular Sludge with Enriched Sulfate and Nitrate.

PubMed

An, Chunjiang; Shi, Yarong; He, Yanling; Huang, Guohe; Liu, Yonghong; Yang, Shucheng

2017-05-01

  RDX and HMX are widely used energetic materials and they are recognized as environmental contaminants at numerous locations. The present study investigated the biotransformation of RDX and HMX by anaerobic granular sludge under sulfate- and nitrate-enriched conditions. The results showed that RDX and HMX could be transformed by anaerobic granular sludge when nitrate was present. However, the biotransformation of RDX and HMX was negatively influenced, especially with high nitrate concentrations. Sulfate-enriched conditions were more favorable for the removal of ammunition compounds by anaerobic granular sludge than nitrate-enriched conditions. The removal of RDX and HMX under both nitrate- and sulfate-enriched conditions was facilitated by the use of glucose as additional substrate. This knowledge may help identify factors required for rapid removal of RDX and HMX in high-rate bioreactors. These results can also be applied to devise an appropriate and practical biological treatment strategy for explosive contaminated wastewater.

Nitrate-nitrogen contamination in groundwater: Spatiotemporal variation and driving factors under cropland in Shandong Province, China

NASA Astrophysics Data System (ADS)

Liu, J.; Jiang, L. H.; Zhang, C. J.; Li, P.; Zhao, T. K.

2017-08-01

High groundwater nitrate-N is a serious problem especially in highly active agricultural areas. In study, the concentration and spatialtemporal distribution of groundwater nitrate-N under cropland in Shandong province were assessed by statistical and geostatistical techniques. Nitrate-N concentration reached a maximum of 184.60 mg L-1 and 29.5% of samples had levels in excess of safety threshold concentration (20 mg L-1). The median nitrate-N contents after rainy season were significantly higher than those before rainy season, and decreased with increasing groundwater depth. Nitrate-N under vegetable and orchard area are significantly higher than ones under grain. The kriging map shows that groundwater nitrate-N has a strong spatial variability. Many districts, such as Weifang, Linyi in Shandong province are heavily contaminated with nitrate-N. However, there are no significant trends of NO3 --N for most cities. Stepwise regression analysis showed influencing factors are different for the groundwater in different depth. But overall, vegetable yield per unit area, percentages of orchard area, per capita agricultural production, unit-area nitrogen fertilizer, livestock per unit area, percentages of irrigation areas, population per unit area and annual mean temperature are significant variables for groundwater nitrate-N variation.

Recurrent diarrhea in children living in areas with high levels of nitrate in drinking water.

PubMed

Gupta, S K; Gupta, R C; Gupta, A B; Seth, A K; Bassin, J K; Gupta, A; Sharma, M L

2001-01-01

Given that there was documented evidence of an association between diarrhea and high nitrate ingestion, the authors examined drinking water nitrate concentration and its possible correlation(s) with methemoglobin levels, cytochrome b5 reductase activity, and recurrent diarrhea. In addition, the authors studied histopathological changes in the intestines of rabbits in an animal model. Five village areas were studied, and nitrate concentrations (expressed in mg of nitrate per liter of water) of 26, 45, 95, 220, and 459 existed in the respective villages. The study included 88 randomly selected children who were 8 yr of age or younger; they represented 10% of the total population of each of the areas. Detailed histories of recurrent diarrhea were noted, and medical examinations were conducted. Cytochrome b5 reductase activity and methemoglobin levels were estimated biochemically. Collected data were analyzed statistically with Microsoft Excel software. In addition, the authors exposed rabbits to various levels of nitrate, and histopathological changes of the stomach and intestine (small and large) were evaluated. There was a strong relationship between nitrate concentration and recurrent diarrhea; 80% of the recurrent diarrhea cases were explained by nitrate concentration alone. In the rabbit intestines, lymphocytic infiltration and hyperplasia characterized the submucosa as nitrate concentrations increased.

Geohydrology, water quality, and nitrogen geochemistry in the saturated and unsaturated zones beneath various land uses, Riverside and San Bernardino counties, California, 1991-93

USGS Publications Warehouse

Rees, Terry F.; Bright, Daniel J.; Fay, Ronald G.; Christensen, Allen H.; Anders, Robert; Baharie, Brian S.; Land, Michael T.

1995-01-01

The U.S. Geological Survey, in cooperation with the Eastern Municipal Water District, the Metropolitan Water District of Southern California, and the Orange County Water District, has completed a detailed study of the Hemet groundwater basin. The quantity of ground water stored in the basin in August 1992 is estimated to be 327,000 acre-feet. Dissolved-solids concentration ranged from 380 to 700 mg/L (milligrams per liter), except in small areas where the concentration exceeded 1,000 mg/L. Nitrate concentrations exceeded the U.S. Environmental Protection Agency Maximum Contaminant Level (MCL) of 10 mg/L nitrate (as nitrogen) in the southeastern part of the basin, in the Domenigoni Valley area, and beneath a dairy in the Diamond Valley area. Seven sites representing selected land uses-- residential, turf grass irrigated with reclaimed water, citrus grove, irrigated farm, poultry farm, and dairy (two sites)--were selected for detailed study of nitrogen geochemistry in the unsaturated zone. For all land uses, nitrate was the dominant nitrogen species in the unsaturated zone.Although nitrate was seasonally present in the shallow unsaturated zone beneath the residential site, it was absent at moderate depths, suggesting negligible migration of nitrate from the surface at this time. Microbial denitrification probably is occurring in the shallow unsaturated zone. High nitrate concentrations in the deep unsaturated zone (greater than 100 ft) suggest either significantly higher nitrate loading at some time in the past, or lateral movement of nitrate at depth. Nitrate also is seasonally present in the shallow unsaturated zone beneath the reclaimed-water site, and (in contrast with the residential site), nitrate is perennially present in the deeper unsaturated zone. Microbial denitrification in the unsaturated zone and in the capillary fringe above the water table decreases the concentrations of nitrate in pore water to below the MCL before reaching the water table.Pore water in the unsaturated zone beneath the citrus grove site contains very high concentrations of nitrate. Even though there are zones of microbial denitrification, nitrate seems to be migrating downward to the water table. The presence of a shallow perched-water zone beneath the irrigated-farm site prevents the vertical movement of nitrate from the surface to the regional water table. Above the perched zone, nitrate concentrations in the unsaturated zone are variable, ranging from below the MCL to four times the MCL. Periodically, nitrate is flushed from the shallow unsaturated zone to the perched-water zone. The unsaturated zone pore-moisture quality could not be adequately addressed because of the very dry conditions in the unsaturated zone beneath the poultry-farm site. Surficial clay deposits prevent water from percolating downward.At the two dairy sites, nitrate loading in pore water at the surface was very high, as great as 7,000 mg/L. Microbial denitrification in the unsaturated zone causes such concentrations to decrease rapidly with depth. At a depth of 20 ft, nitrate concentration was less than 100 mg/L. In areas where the depth to water is less than 20 ft, nitrate loading to ground water can be very high, whereas in areas where depth to water is greater than 100 ft, most of the nitrate is microbially removed before reaching the water table.

Real time in situ detection of organic nitrates in atmospheric aerosols

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rollins, Andrew W.; Smith, Jared D.; Wilson, Kevin R.

2010-06-11

A new field instrument is described that quantifies total particle phase organic nitrates. The instrument is based on the thermal dissociation laser induced fluorescence (TD-LIF) method that thermally converts nitrates to NO2 which is then detected by LIF. This instrument is unique in its ability to provide fast sensitive measurements of particle phase organic nitrates, without interference from inorganic nitrate. Here we use it to quantify organic nitrates in SOA generated from high-NOx photooxidation of limonene, a-pinene, D-3-carene, and tridecane. In these experiments the organic nitrate moiety is observed to be 6-15percent of the total SOA mass, depending on themore » organic precursor.« less

Ingested Nitrate and Breast Cancer in the Spanish Multicase-Control Study on Cancer (MCC-Spain)

PubMed Central

Espejo-Herrera, Nadia; Gracia-Lavedan, Esther; Pollan, Marina; Aragonés, Nuria; Boldo, Elena; Perez-Gomez, Beatriz; Altzibar, Jone M.; Amiano, Pilar; Zabala, Ana Jiménez; Ardanaz, Eva; Guevara, Marcela; Molina, Antonio J.; Barrio, Juan Pablo; Gómez-Acebo, Ines; Tardón, Adonina; Peiró, Rosana; Chirlaque, Maria Dolores; Palau, Margarita; Muñoz, Montse; Font-Ribera, Laia; Castaño-Vinyals, Gemma; Kogevinas, Manolis; Villanueva, Cristina M.

2016-01-01

Background: Ingested nitrate leads to endogenous formation of N-nitroso compounds that are breast carcinogens in animals, but human evidence is limited. Objective: We evaluated ingested nitrate as a risk factor for breast cancer (BC) in a multicase–control study. Methods: Hospital-based incident BC cases and population-based controls were recruited in eight Spanish regions in 2008–2013; participants provided residential and water consumption from 18 years of age and information on known BC risk factors. Long-term nitrate levels (1940–2010) were estimated and linked with residential histories and water consumption to calculate waterborne ingested nitrate (milligrams/day). Dietary ingested nitrate (milligrams/day) was calculated using food frequency questionnaires and published dietary nitrate contents. Interactions with endogenous nitrosation factors and other variables were evaluated. A total of 1,245 cases and 1,520 controls were included in the statistical analysis. Results: Among the study regions, average ± SD waterborne ingested nitrate ranged from 2.9 ± 1.9 to 13.5 ± 7.5 mg/day, and dietary ingested nitrate ranged from 88.5 ± 48.7 to 154 ± 87.8 mg/day. Waterborne ingested nitrate was not associated with BC overall, but among postmenopausal women, those with both high nitrate (> 6 vs. < 2.6 mg/day) and high red meat intake (≥ 20 vs. < 20 g/day) were more likely to be cases than women with low nitrate and low red meat intake (adjusted odds ratio = 1.64; 95% confidence interval: 1.08, 2.49; overall interaction p-value = 0.17). No association was found with dietary nitrate. Conclusions: Waterborne ingested nitrate was associated with BC only among postmenopausal women with high red meat consumption. Dietary nitrate was not associated with BC regardless of the animal or vegetable source or of menopausal status. Citation: Espejo-Herrera N, Gracia-Lavedan E, Pollan M, Aragonés N, Boldo E, Perez-Gomez B, Altzibar JM, Amiano P, Zabala AJ, Ardanaz E, Guevara M, Molina AJ, Barrio JP, Gómez-Acebo I, Tardón A, Peiró R, Chirlaque MD, Palau M, Muñoz M, Font-Ribera L, Castaño-Vinyals G, Kogevinas M, Villanueva CM. 2016. Ingested nitrate and breast cancer in the Spanish Multicase-Control Study on Cancer (MCC-Spain). Environ Health Perspect 124:1042–1049; http://dx.doi.org/10.1289/ehp.1510334 PMID:26942716

Cylodextrin Polymer Nitrate

NASA Technical Reports Server (NTRS)

Kosowski, Bernard; Ruebner, Anja; Statton, Gary; Robitelle, Danielle; Meyers, Curtis

2000-01-01

The development of the use of cyclodextrin nitrates as possible components of insensitive, high-energy energetics is outlined over a time period of 12 years. Four different types of cyclodextrin polymers were synthesized, nitrated, and evaluated regarding their potential use for the military and aerospace community. The synthesis of these novel cyclodextrin polymers and different nitration techniques are shown and the potential of these new materials is discussed.

A Multi-Tracer Approach to Characterize Sources and Transport of Nitrate in Groundwater in Mantled Karst, Northern Florida

NASA Astrophysics Data System (ADS)

Katz, B. G.; Bohlke, J.; Hornsby, D.

2001-05-01

Nitrate is readily transported from agricultural activities at the surface to the Upper Floridan aquifer in northern Florida due to karst features mantled by highly permeable sands and a high recharge rate (50 cm/yr). In Suwannee and Lafayette Counties, nitrate contamination of groundwater is widespread due to the 10-30 kg/ha nitrogen (N) applied annually for the past few decades as synthetic fertilizers (the dominant source of N). Water samples were collected from 12 springs during baseflow conditions (1997-99) and monthly from 14 wells (1998-99). Springwaters were analyzed for various chemical (N species, dissolved gases, CFCs) and isotopic tracers (15N, 3H/3He, 18O, D, 13C). Water from wells was analyzed monthly for N species, and during low-flow and high-flow conditions for 15N, 18O, D, and 13C. As a result of oxic conditions in the aquifer, nitrate was the dominant N species in water samples. Large monthly fluctuations of groundwater nitrate concentrations were observed at most wells. Relatively high nitrate concentrations in groundwater from 7 wells likely resulted from seasonal agricultural practices including fertilizer applications and manure spreading on cropland. Relatively low nitrate concentrations in groundwater from two wells during high-flow conditions were related to mixing with river water. Groundwater samples had N-isotope values (3.8-11.7 per mil) that indicated varying mixtures of inorganic and organic N sources, which corresponded in part to varying proportions of synthetic fertilizers and manure applied to fields. In springwaters from Suwannee County, nitrate trends and N-isotope data (2.7-6.2 per mil) were consistent with a peak in fertilizer N input in the late 1970's and a relatively high overall ratio of artificial fertilizer/manure. In contrast, springwater nitrate trends and N-isotope data (4.5-9.1 per mil) in Lafayette County were consistent with a more monotonic increase in fertilizer N input and relatively low overall ratio of artificial fertilizer/manure. Dampened nitrate trends in springwaters in both counties, relative to trends in estimated N inputs, likely were related to ages of groundwater discharging from springs that are on the order of decades (10-30 years), based on 3H/3He and CFC age-dating techniques.

Thermophilic nitrate-reducing microorganisms prevent sulfate reduction in cold marine sediments incubated at high temperature

NASA Astrophysics Data System (ADS)

Nepomnyashchaya, Yana; Rezende, Julia; Hubert, Casey

2014-05-01

Hydrogen sulphide produced during metabolism of sulphate-reducing microorganisms (SRM) is toxic, corrosive and causes detrimental oil reservoir souring. During secondary oil recovery, injecting oil reservoirs with seawater that is rich in sulphate and that also cools high temperature formations provides favourable growth conditions for SRM. Nitrate addition can prevent metabolism of SRM by stimulating nitrate-reducing microorganisms (NRM). The investigations of thermophilic NRM are needed to develop mechanisms to control the metabolism of SRM in high temperature oil field ecosystems. We therefore established a model system consisting of enrichment cultures of cold surface marine sediments from the Baltic Sea (Aarhus Bay) that were incubated at 60°C. Enrichments contained 25 mM nitrate and 40 mM sulphate as potential electron acceptors, and a mixture of the organic substrates acetate, lactate, propionate, butyrate (5 mM each) and yeast extract (0.01%) as potential carbon sources and electron donors. Slurries were incubated at 60°C both with and without initial pasteurization at 80°C for 2 hours. In the enrichments containing both nitrate and sulphate, the concentration of nitrate decreased indicating metabolic activity of NRM. After a four-hour lag phase the rate of nitrate reduction increased and the concentration of nitrate dropped to zero after 10 hours of incubation. The concentration of nitrite increased as the reduction of nitrate progressed and reached 16.3 mM after 12 hours, before being consumed and falling to 4.4 mM after 19-day of incubation. No evidence for sulphate reduction was observed in these cultures during the 19-day incubation period. In contrast, the concentration of sulphate decreased up to 50% after one week incubation in controls containing only sulphate but no nitrate. Similar sulfate reduction rates were seen in the pasteurized controls suggesting the presence of heat resistant SRM, whereas nitrate reduction rates were lower in the pasteurized experiment, suggesting either different populations of NRM or a population of NRM that was not resistant to the 80°C pre-treatment. These results demonstrate that thermophilic NRM exist in cold marine sediments from Aarhus Bay and can be enriched under appropriate conditions. Effective microbial control of SRM activity at high temperature in our Aarhus Bay sediment model system depends on the addition of nitrate to stimulate this group of microorganisms.

Concurrent microbial reduction of high concentrations of nitrate and perchlorate in an ion exchange membrane bioreactor.

PubMed

Fox, Shalom; Bruner, Tali; Oren, Yoram; Gilron, Jack; Ronen, Zeev

2016-09-01

We investigated effective simultaneous removal of high loads of nitrate and perchlorate from synthetic groundwater using an ion exchange membrane bioreactor (IEMB). The aim of this research was to characterize both transport aspects and biodegradation mechanisms involved in the treatment process of high loads of the two anions. Biodegradation process was proven to be efficient with over 99% efficiency of both perchlorate and nitrate, regardless of their load. The maximum biodegradation rates were 18.3 (mmol m(-2)  h(-1) ) and 5.5 (mmol m(-2)  h(-1) ) for nitrate and perchlorate, respectively. The presence of a biofilm on the bio-side of the membrane only slightly increased the nitrate and perchlorate transmembrane flux as compared to the measured flux during a Donnan dialysis experiment where there is no biodegradation of perchlorate and nitrate in the bio-compartment. The nitrate flux in presence of a biofilm was 18.3 (±1.9) (mmole m(-2)  h(-1) ), while without the biofilm, the flux was 16.9 (±1.5) (mmole m(-2)  h(-1) ) for the same feed inlet nitrate concentration of 4 mM. The perchlorate transmembrane flux increased similarly by an average of 5%. Samples of membrane biofilm and suspended bacteria from the bio-reactor were analyzed for diversity and abundance of the perchlorate and nitrate reducing bacteria. Klebsiella oxytoca, known as a glycerol fermenter, accounted for 70% of the suspended bacteria. In contrast, perchlorate and nitrate reducing bacteria predominated in the biofilm present on the membrane. These results are consistent with our proposed two stage biodegradation mechanism where glycerol is first fermented in the suspended phase of the bio-reactor and the fermentation products drive perchlorate and nitrate bio-reduction in the biofilm attached to the membrane. These results suggest that the niche exclusion of microbial populations in between the reactor and membrane is controlled by the fluxes of the electron donors and acceptors. Such a mechanism has important implications for controlling the bio-reduction reaction in the IEMB when using glycerol as a carbon source and allowing treating a complex contamination of high concentrations of perchlorate and nitrating in groundwater and successfully biodegrading them to non-hazardous components. Biotechnol. Bioeng. 2016;113: 1881-1891. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Coniferous coverage as well as catchment steepness influences local stream nitrate concentrations within a nitrogen-saturated forest in central Japan.

PubMed

Watanabe, Mirai; Miura, Shingo; Hasegawa, Shun; Koshikawa, Masami K; Takamatsu, Takejiro; Kohzu, Ayato; Imai, Akio; Hayashi, Seiji

2018-04-28

High concentrations of nitrate have been detected in streams flowing from nitrogen-saturated forests; however, the spatial variations of nitrate leaching within those forests and its causes remain poorly explored. The aim of this study is to evaluate the influences of catchment topography and coniferous coverage on stream nitrate concentrations in a nitrogen-saturated forest. We measured nitrate concentrations in the baseflow of headwater streams at 40 montane forest catchments on Mount Tsukuba in central Japan, at three-month intervals for 1 year, and investigated their relationship with catchment topography and with coniferous coverage. Although stream nitrate concentrations varied from 0.5 to 3.0 mgN L -1 , those in 31 catchments consistently exceeded 1 mgN L -1 , indicating that this forest had experienced nitrogen saturation. A classification and regression tree analysis with multiple environmental factors showed that the mean slope gradient and coniferous coverage were the best and second best, respectively, at explaining inter-catchment variance of stream nitrate concentrations. This analysis suggested that the catchments with steep topography and high coniferous coverage tend to have high nitrate concentrations. Moreover, in the three-year observation period for five adjacent catchments, the two catchments with relatively higher coniferous coverage consistently had higher stream nitrate concentrations. Thus, the spatial variations in stream nitrate concentrations were primarily regulated by catchment steepness and, to a lesser extent, coniferous coverage in this nitrogen-saturated forest. Our results suggest that a decrease in coniferous coverage could potentially contribute to a reduction in nitrate leaching from this nitrogen-saturated forest, and consequently reduce the risk of nitrogen overload for the downstream ecosystems. This information will allow land managers and researchers to develop improved management plans for this and similar forests in Japan and elsewhere. Copyright © 2018 Elsevier B.V. All rights reserved.

Variation of isomer distribution in electrophilic nitration of toluene, anisole, and o-xylene: Independence of high regioselectivity from reactivity of reagent*

PubMed Central

Olah, George A.; Lin, Henry C.; Olah, Judith A.; Narang, Subhash C.

1978-01-01

The nitration of toluene and anisole was studied with nitrating systems of varying reactivity. High regioselectivity of ortho-para over meta substitution was maintained in all nitrations, regardless of the reactivity of the nitrating system. At the same time, the amount of meta substitution stayed low (3% or less), even when the fast reactions may have reached the encounter-controlled limit. Because the nitration of o-xylene, in which both ring positions are activated by the effect of a methyl group, also does not show any diminishing of regioselectivity, the possibility of a dual mechanistic pathway, in which the activated position would react by a fast, encounter-controlled path, whereas the nonactivated meta position by a slower σ-type path, can be ruled out. The data unambiguously prove that the high regioselectivity of electrophilic aromatic nitration is independent of the reactivity of the reagent, because no significant increase of meta substitution of toluene or anisole was observed, regardless of the activity of the nitrating system. No selectivity-reactivity relationship is thus evident and the ortho-para directing effect of primary substituents over meta substitution is always maintained. The variation in the amount of the meta isomer, up to the observed limit of about 3% in the case of toluene and <2% for anisole, is probably significant but, at the present time, cannot be quantitatively evaluated with the ±0.5% overall reproducible accuracy of the nitrations. Steric factors, such as increasing bulkiness of the nitrating agent, also can affect the ortho-para isomer ratios but are not considered to be the only reason for the observed variations, which reflect the specific nitrating systems, affecting the nature and position of the transition state of highest energy on the reaction pathway. PMID:16592489

Nitrate leaching in a winter wheat-summer maize rotation on a calcareous soil as affected by nitrogen and straw management

PubMed Central

Huang, Tao; Ju, Xiaotang; Yang, Hao

2017-01-01

Nitrate leaching is one of the most important pathways of nitrogen (N) loss which leads to groundwater contamination or surface water eutrophication. Clarifying the rates, controlling factors and characteristics of nitrate leaching is the pre-requisite for proposing effective mitigation strategies. We investigated the effects of interactions among chemical N fertilizer, straw and manure applications on nitrogen leaching in an intensively managed calcareous Fluvo-aquic soil with winter wheat-summer maize cropping rotations on the North China Plain from October 2010 to September 2013 using ceramic suction cups and seepage water calculations based on a long-term field experiment. Annual nitrate leaching reached 38–60 kg N ha−1 from conventional N managements, but declined by 32–71% due to optimum N, compost manure or municipal waste treatments, respectively. Nitrate leaching concentrated in the summer maize season, and fewer leaching events with high amounts are the characteristics of nitrate leaching in this region. Overuse of chemical N fertilizers, high net mineralization and nitrification, together with predominance of rainfall in the summer season with light soil texture are the main controlling factors responsible for the high nitrate leaching loss in this soil-crop-climatic system. PMID:28176865

Adaptation of cytochrome-b5 reductase activity and methaemoglobinaemia in areas with a high nitrate concentration in drinking-water.

PubMed Central

Gupta, S. K.; Gupta, R. C.; Seth, A. K.; Gupta, A. B.; Bassin, J. K.; Gupta, A.

1999-01-01

An epidemiological investigation was undertaken in India to assess the prevalence of methaemoglobinaemia in areas with high nitrate concentration in drinking-water and the possible association with an adaptation of cytochrome-b5 reductase. Five areas were selected, with average nitrate ion concentrations in drinking-water of 26, 45, 95, 222 and 459 mg/l. These areas were visited and house schedules were prepared in accordance with a statistically designed protocol. A sample of 10% of the total population was selected in each of the areas, matched for age and weight, giving a total of 178 persons in five age groups. For each subject, a detailed history was documented, a medical examination was conducted and blood samples were taken to determine methaemoglobin level and cytochrome-b5 reductase activity. Collected data were subjected to statistical analysis to test for a possible relationship between nitrate concentration, cytochrome-b5 reductase activity and methaemoglobinaemia. High nitrate concentrations caused methaemoglobinaemia in infants and adults. The reserve of cytochrome-b5 reductase activity (i.e. the enzyme activity not currently being used, but which is available when needed; for example, under conditions of increased nitrate ingestion) and its adaptation with increasing water nitrate concentration to reduce methaemoglobin were more pronounced in children and adolescents. PMID:10534899

The ability to use nitrate confers advantage to Dekkera bruxellensis over S. cerevisiae and can explain its adaptation to industrial fermentation processes.

PubMed

de Barros Pita, Will; Leite, Fernanda Cristina Bezerra; de Souza Liberal, Anna Theresa; Simões, Diogo Ardaillon; de Morais, Marcos Antonio

2011-06-01

The yeast Dekkera bruxellensis has been regarded as a contamination problem in industrial ethanol production because it can replace the originally inoculated Saccharomyces cerevisiae strains. The present study deals with the influence of nitrate on the relative competitiveness of D. bruxellensis and S. cerevisiae in sugar cane ethanol fermentations. The industrial strain D. bruxellensis GDB 248 showed higher growth rates than S. cerevisiae JP1 strain in mixed ammonia/nitrate media, and nitrate assimilation genes were only slightly repressed by ammonia. These characteristics rendered D. bruxellensis cells with an ability to overcome S. cerevisiae populations in both synthetic medium and in sugar cane juice. The results were corroborated by data from industrial fermentations that showed a correlation between high nitrate concentrations and high D. bruxellensis cell counts. Moreover, the presence of nitrate increased fermentation efficiency of D. bruxellensis cells in anaerobic conditions, which may explain the maintenance of ethanol production in the presence of D. bruxellensis in industrial processes. The presence of high levels of nitrate in sugar cane juice may be due to its inefficient conversion by plant metabolism in certain soil types and could explain the periodical episodes of D. bruxellensis colonization of Brazilian ethanol plants.

Unusual seasonal patterns and inferred processes of nitrogen retention in forested headwater catchments of the Upper Susquehanna basin

NASA Astrophysics Data System (ADS)

Goodale, C. L.; Thomas, S. A.; Fredriksen, G.; Elliott, E. M.; Flinn, K. M.; Butler, T. J.

2008-12-01

The Susquehanna River provides two-thirds of the annual nitrogen (N) load to the Chesapeake Bay, and atmospheric deposition is a major contributor to the basin's N inputs. Yet, there are few measurements of the retention of atmospheric N in the Upper Susquehanna's forested headwaters. We characterized the amount, form (nitrate, ammonium, and dissolved organic nitrogen), isotopic composition (del18O- and del15N-nitrate), and seasonality of stream N over two years from 8-15 small forested headwater catchments of the Susquehanna Basin. We expected high rates of N retention and seasonal nitrate patterns typical of other seasonally snow-covered catchments: dormant season peaks and growing season minima. Annual nitrate exports were approximately 0.1-0.7 kg N ha-1 y-1, and correlated positively with the percent of catchment free from historical agriculture. DON export averaged 0.6 +/- 0.1 kg N ha-1 y-1. All catchments had high rates of N retention but with atypical seasonal nitrate patterns, consisting of summer peaks, fall crashes, and modest rebounds during the dormant season. The fall nitrate crash coincided with carbon inputs at leaffall, indicating in-stream heterotrophic uptake. Stream del18O-nitrate values indicated microbial nitrification as the dominant source of stream nitrate, with modest contributions directly from precipitation in early stages of snowmelt. Three hypothesized sources of summer nitrate peaks include: delayed release of nitrate flushed to groundwater at snowmelt, weathering of geologic N, and increased net nitrate production. Measurements of shale del15N as well as soil, well-, and springwater nitrate within one catchment point toward a summer increase in net nitrification in surface soils. Rather than plant demand, processes governing the production, retention, and hydrologic transport of nitrate in surface mineral soils may drive the unusual nitrate seasonality in this and other systems, and provide insights on N retention in general.

Nitrate consumption in sediments of the German Bight (North Sea)

NASA Astrophysics Data System (ADS)

Neumann, Andreas; van Beusekom, Justus E. E.; Holtappels, Moritz; Emeis, Kay-Christian

2017-09-01

Denitrification on continental margins and in coastal sediments is a major sink of reactive N in the present nitrogen cycle and a major ecosystem service of eutrophied coastal waters. We analyzed the nitrate removal in surface sediments of the Elbe estuary, Wadden Sea, and adjacent German Bight (SE North Sea) during two seasons (spring and summer) along a eutrophication gradient ranging from a high riverine nitrate concentrations at the Elbe Estuary to offshore areas with low nitrate concentrations. The gradient encompassed the full range of sediment types and organic carbon concentrations of the southern North Sea. Based on nitrate penetration depth and concentration gradient in the porewater we estimated benthic nitrate consumption rates assuming either diffusive transport in cohesive sediments or advective transport in permeable sediments. For the latter we derived a mechanistic model of porewater flow. During the peak nitrate discharge of the river Elbe in March, the highest rates of diffusive nitrate uptake were observed in muddy sediments (up to 2.8 mmol m- 2 d- 1). The highest advective uptake rate in that period was observed in permeable sediment and was tenfold higher (up to 32 mmol m- 2 d- 1). The intensity of both diffusive and advective nitrate consumption dropped with the nitrate availability and thus decreased from the Elbe estuary towards offshore stations, and were further decreased during late summer (minimum nitrate discharge) compared to late winter (maximum nitrate discharge). In summary, our rate measurements indicate that the permeable sediment accounts for up to 90% of the total benthic reactive nitrogen consumption in the study area due to the high efficiency of advective nitrate transport into permeable sediment. Extrapolating the averaged nitrate consumption of different sediment classes to the areas of Elbe Estuary, Wadden Sea and eastern German Bight amounts to an N-loss of 3.1 ∗ 106 mol N d- 1 from impermeable, diffusion-controlled sediment, and 5.2 ∗ 107 mol N d- 1 from permeable sediment with porewater advection.

Effect of ammonium and nitrate on ferric chelate reductase and nitrate reductase in Vaccinium species.

PubMed

Poonnachit, U; Darnell, R

2004-04-01

Most Vaccinium species have strict soil requirements for optimal growth, requiring low pH, high iron availability and nitrogen primarily in the ammonium form. These soils are limited and are often located near wetlands. Vaccinium arboreum is a wild species adapted to a wide range of soils, including high pH, low iron, and nitrate-containing soils. This broader soil adaptation in V. arboreum may be related to increased efficiency of iron or nitrate uptake compared with the cultivated Vaccinium species. Nitrate, ammonium and iron uptake, and nitrate reductase (NR) and ferric chelate reductase (FCR) activities were compared in two Vaccinium species grown hydroponically in either nitrate or ammonia, with or without iron. The species studied were the wild V. arboreum and the cultivated V. corymbosum interspecific hybrid, which exhibits the strict soil requirements of most Vaccinium species. Ammonium uptake was significantly greater than nitrate uptake in both species, while nitrate uptake was greater in the wild species, V. arboreum, compared with the cultivated species, V. corymbosum. The increased nitrate uptake in V. arboreum was correlated with increased root NR activity compared with V. corymbosum. The lower nitrate uptake in V. corymbosum was reflected in decreased plant dry weight in this species compared with V. arboreum. Root FCR activity increased significantly in V. corymbosum grown under iron-deficient conditions, compared with the same species grown under iron-sufficient conditions or with V. arboreum grown under either iron condition. V. arboreum appears to be more efficient in acquiring nitrate compared with V. corymbosum, possibly due to increased NR activity and this may partially explain the wider soil adaptation of V. arboreum.

Temperature and Substrate Control Woodchip Bioreactor Performance in Reducing Tile Nitrate Loads in East-Central Illinois.

PubMed

David, Mark B; Gentry, Lowell E; Cooke, Richard A; Herbstritt, Stephanie M

2016-05-01

Tile drainage is the major source of nitrate in the upper Midwest, and end-of-tile removal techniques such as wood chip bioreactors have been installed that allow current farming practices to continue, with nitrate removed through denitrification. There have been few multiyear studies of bioreactors examining controls on nitrate removal rates. We evaluated the nitrate removal performance of two wood chip bioreactors during the first 3 yr of operation and examined the major factors that regulated nitrate removal. Bioreactor 2 was subject to river flooding, and performance was not assessed. Bioreactor 1 had average monthly nitrate removal rates of 23 to 44 g N m d in Year 1, which decreased to 1.2 to 11 g N m d in Years 2 and 3. The greater N removal rates in Year 1 and early in Year 2 were likely due to highly degradable C in the woodchips. Only late in Year 2 and in Year 3 was there a strong temperature response in the nitrate removal rate. Less than 1% of the nitrate removed was emitted as NO. Due to large tile inputs of nitrate (729-2127 kg N) at high concentrations (∼30 mg nitrate N L) in Years 2 and 3, overall removal efficiency was low (3 and 7% in Years 2 and 3, respectively). Based on a process-based bioreactor performance model, Bioreactor 1 would have needed to be 9 times as large as the current system to remove 50% of the nitrate load from this 20-ha field. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

High pre-transplant serum nitrate levels predict risk of acute steroid-refractory graft-versus-host disease in the absence of statin therapy

PubMed Central

Dietrich, Sascha; Okun, Jürgen G.; Schmidt, Kathrin; Falk, Christine S.; Wagner, Andreas H.; Karamustafa, Suzan; Radujkovic, Aleksandar; Hegenbart, Ute; Ho, Anthony D.; Dreger, Peter; Luft, Thomas

2014-01-01

Steroid-refractory graft-versus-host disease is a life-threatening complication after allogeneic stem cell transplantation. Evidence is accumulating that steroid-refractory graft-versus-host disease is associated with endothelial distress. Endothelial cell homeostasis is regulated by nitric oxide, and serum nitrates are derived from nitric oxide synthase activity or dietary sources. In this retrospective study based on 417 patients allografted at our institution we investigated whether quantification of serum nitrates could predict steroid-refractory graft-versus-host disease. Elevated pre-transplant levels of serum nitrates (>26.5 μM) predicted steroid-refractory graft-versus-host disease (P=0.026) and non-relapse mortality (P=0.028), particularly in combination with high pre-transplant angiopoietin-2 levels (P=0.0007 and P=0.021, respectively). Multivariate analyses confirmed serum nitrates as independent predictors of steroid-refractory graft-versus-host disease and non-relapse mortality. Differences in serum nitrate levels did not correlate with serum levels of tumor necrosis factor or C-reactive protein or expression of inducible nitric oxide synthase in blood cells. Patients with high pre-transplant nitrate levels had significantly reduced rates of refractory graft-versus-host disease (P=0.031) when pravastatin was taken. In summary, patients at high risk of developing steroid-refractory graft-versus-host disease could be identified prior to transplantation by serum markers linked to endothelial cell function. Retrospectively, statin medication was associated with a reduced incidence of refractory graft-versus-host disease in this endothelial high-risk cohort. PMID:24142995

Identifying the influential aquifer heterogeneity factor on nitrate reduction processes by numerical simulation

NASA Astrophysics Data System (ADS)

Jang, E.; He, W.; Savoy, H.; Dietrich, P.; Kolditz, O.; Rubin, Y.; Schüth, C.; Kalbacher, T.

2017-01-01

Nitrate reduction reactions in groundwater systems are strongly influenced by various aquifer heterogeneity factors that affect the transport of chemical species, spatial distribution of redox reactive substances and, as a result, the overall nitrate reduction efficiency. In this study, we investigated the influence of physical and chemical aquifer heterogeneity, with a focus on nitrate transport and redox transformation processes. A numerical modeling study for simulating coupled hydrological-geochemical aquifer heterogeneity was conducted in order to improve our understanding of the influence of the aquifer heterogeneity on the nitrate reduction reactions and to identify the most influential aquifer heterogeneity factors throughout the simulation. Results show that the most influential aquifer heterogeneity factors could change over time. With abundant presence of electron donors in the high permeable zones (initial stage), physical aquifer heterogeneity significantly influences the nitrate reduction since it enables the preferential transport of nitrate to these zones and enhances mixing of reactive partners. Chemical aquifer heterogeneity plays a comparatively minor role. Increasing the spatial variability of the hydraulic conductivity also increases the nitrate removal efficiency of the system. However, ignoring chemical aquifer heterogeneity can lead to an underestimation of nitrate removals in long-term behavior. With the increase of the spatial variability of the electron donor, i.e. chemical heterogeneity, the number of the ;hot spots; i.e. zones with comparably higher reactivity, should also increase. Hence, nitrate removal efficiencies will also be spatially variable but overall removal efficiency will be sustained if longer time scales are considered and nitrate fronts reach these high reactivity zones.

Protein tyrosine phosphatase 1B inhibitory activity of alkaloids from Rhizoma Coptidis and their molecular docking studies.

PubMed

Choi, Jae Sue; Ali, Md Yousof; Jung, Hyun Ah; Oh, Sang Ho; Choi, Ran Joo; Kim, Eon Ji

2015-08-02

Rhizoma Coptidis (the rhizome of Coptis chinensis Franch) has commonly been used for treatment of diabetes mellitus in traditional Chinese medicine due to its blood sugar-lowering properties and therapeutic benefits which highly related to the alkaloids therein. However, a limited number of studies focused on the Coptis alkaloids other than berberine. In the present study, we investigated the anti-diabetic potential of Coptis alkaloids, including berberine (1), epiberberine (2), magnoflorine (3), and coptisine (4), by evaluating the ability of these compounds to inhibit protein tyrosine phosphatase 1B (PTP1B), and ONOO(-)-mediated protein tyrosine nitration. We scrutinized the potentials of Coptis alkaloids as PTP1B inhibitors via enzyme kinetics and molecular docking simulation. The Coptis alkaloids 1-4 exhibited remarkable inhibitory activities against PTP1B with the IC50 values of 16.43, 24.19, 28.14, and 51.04 μM, respectively, when compared to the positive control ursolic acid. These alkaloids also suppressed ONOO(-)-mediated tyrosine nitration effectively in a dose dependent manner. In addition, our kinetic study using the Lineweaver-Burk and Dixon plots revealed that 1 and 2 showed a mixed-type inhibition against PTP1B, while 3 and 4 noncompetitively inhibited PTP1B. Moreover, molecular docking simulation of these compounds demonstrated negative binding energies (Autodock 4.0=-6.7 to -7.8 kcal/mol; Fred 2.0=-59.4 to -68.2 kcal/mol) and a high proximity to PTP1B residues, including Phe182 and Asp181 in the WPD loop, Cys215 in the active sites and Tyr46, Arg47, Asp48, Val49, Ser216, Ala217, Gly218, Ile219, Gly220, Arg221 and Gln262 in the pocket site, indicating a higher affinity and tighter binding capacity of these alkaloids for the active site of the enzyme. Our results clearly indicate the promising anti-diabetic potential of Coptis alkaloids as inhibitors on PTP1B as well as suppressors of ONOO(-)-mediated protein tyrosine nitration, and thus hold promise as therapeutic agents for the treatment of diabetes and related disease. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Long-term tillage and crop rotation effects on residual nitrate in the crop root zone and nitrate accumulation in the intermediate vadose zone

USGS Publications Warehouse

Katupitiya, A.; Eisenhauer, D.E.; Ferguson, R.B.; Spalding, R.F.; Roeth, F.W.; Bobier, M.W.

1997-01-01

Tillage influences the physical and biological environment of soil. Rotation of crops with a legume affects the soil N status. A furrow irrigated site was investigated for long-term tillage and crop rotation effects on leaching of nitrate from the root zone and accumulation in the intermediate vadose zone (IVZ). The investigated tillage systems were disk-plant (DP), ridge-till (RT) and slot-plant (SP). These tillage treatments have been maintained on the Hastings silt loam (Udic Argiustoll) and Crete silt loam (Pachic Argiustoll) soils since 1976. Continuous corn (CC) and corn soybean (CS) rotations were the subtreatments. Since 1984, soybeans have been grown in CS plots in even calendar years. All tillage treatments received the same N rate. The N rate varied annually depending on the root zone residual N. Soybeans were not fertilized with N-fertilizer. Samples for residual nitrate in the root zone were taken in 8 of the 15 year study while the IVZ was only sampled at the end of the study. In seven of eight years, root zone residual soil nitrate-N levels were greater with DP than RT and SP. Residual nitrate-N amounts were similar in RT and SP in all years. Despite high residual nitrate-N with DP and the same N application rate, crop yields were higher in RT and SP except when DP had an extremely high root zone nitrate level. By applying the same N rates on all tillage treatments, DP may have been fertilized in excess of crop need. Higher residual nitrate-N in DP was most likely due to a combination of increased mineralization with tillage and lower yield compared to RT and SP. Because of higher nitrate availability with DP, the potential for nitrate leaching from the root zone was greater with DP as compared to the RT and SP tillage systems. Spring residual nitrate-N contents of DP were larger than RT and SP in both crop rotations. Ridge till and SP systems had greater nitrate-N with CS than CC rotations. Nitrate accumulation in IVZ at the upstream end of the field was twice as high with DP compared to RT and SP. At the downstream end, it was 2.4 and 1.6 times greater with DP than RT and SP, respectively. Nitrate concentration was greater in the IVZ of DP compared to RT and SP tillage systems. Nitrate accumulations in IVZ of RT and SP were not different. Continuous corn had slightly higher nitrate levels in IVZ than CS. The depth of nitrate penetration at the upstream end was greater than that of the downstream end. Estimated rates of nitrate movement ranged from 0.87 to 0.92 m yr-1 at the upstream end and 0.73 to 0.78 m yr-1 at the downstream end.

Histological characteristics following a long-term nitrate-rich diet in miniature pigs with parotid atrophy

PubMed Central

Xia, Dengsheng; Qu, Xingmin; Tran, Simon D; Schmidt, Laura L; Qin, Lizheng; Zhang, Chunmei; Cui, Xiuyu; Deng, Dajun; Wang, Songlin

2015-01-01

The aim of this study was to investigate the histological characteristics following a 2-year nitrate-rich diet in miniature pigs with parotid atrophy. Using averages collected data from three time points at 6, 12, and 24 months following the induction of parotid gland atrophy, salivary nitrate levels of the nitrate-diet parotid-atrophied group (17.3±3.9 ng/µl) were close to those of the control group (19.6±5.1 ng/µl). Compared to the control group, the nitrate-diet group had significantly higher nitrate levels in blood (P < 0.05) and urine (P < 0.001). Histological and electron microscopy analyses showed no abnormalities in the organs of experimental or control animals. No significant differences on apoptosis rate were found in liver and kidney tissues between the standard- and nitrate-diet groups. Therefore, dietary nitrate supplementation could restore salivary nitrate levels. High-dose nitrate loading for 2 years had no observed systemic toxicity in miniature pigs with parotid atrophy. PMID:26261499

IgG1 memory B cells keep the memory of IgE responses.

PubMed

He, Jin-Shu; Subramaniam, Sharrada; Narang, Vipin; Srinivasan, Kandhadayar; Saunders, Sean P; Carbajo, Daniel; Wen-Shan, Tsao; Hidayah Hamadee, Nur; Lum, Josephine; Lee, Andrea; Chen, Jinmiao; Poidinger, Michael; Zolezzi, Francesca; Lafaille, Juan J; Curotto de Lafaille, Maria A

2017-09-21

The unique differentiation of IgE cells suggests unconventional mechanisms of IgE memory. IgE germinal centre cells are transient, most IgE cells are plasma cells, and high affinity IgE is produced by the switching of IgG1 cells to IgE. Here we investigate the function of subsets of IgG1 memory B cells in IgE production and find that two subsets of IgG1 memory B cells, CD80 + CD73 + and CD80 - CD73 - , contribute distinctively to the repertoires of high affinity pathogenic IgE and low affinity non-pathogenic IgE. Furthermore, repertoire analysis indicates that high affinity IgE and IgG1 plasma cells differentiate from rare CD80 + CD73 + high affinity memory clones without undergoing further mutagenesis. By identifying the cellular origin of high affinity IgE and the clonal selection of high affinity memory B cells into the plasma cell fate, our findings provide fundamental insights into the pathogenesis of allergies, and on the mechanisms of antibody production in memory B cell responses.IgE is an important mediator of protective immunity as well as allergic reaction, but how high affinity IgE antibodies are produced in memory responses is not clear. Here the authors show that IgE can be generated via class-switch recombination in IgG1 memory B cells without additional somatic hypermutation.

Nitrate and drinking water from private wells: will there be an epidemic of cancers of the digestive tract, urinary bladder and thyroid?

PubMed

Njeze, G E; Dilibe, U; Ilo, C

2014-01-01

To estimate the nitrate levels in private wells located in different parts of Enugu and discuss the future health implications following chronic ingestion of well water. The map of Enugu was used to divide the city into many 25 units, using grid lines 1 cm apart. Cluster sampling method was used to collect samples. These samples were sent to two laboratories for estimation of nitrate levels. The people drawing water from the different wells were interviewed to determine what they used the water for. The subjects who were interviewed said they ingested the water. The nitrate levels found in these wells (median value of 31 mg/L) were significantly higher than the internationally accepted levels of nitrate in water for ingestion, (P < 0.0001). High nitrate levels drinking water is dangerous to health and can cause methemoglobinemia in children. It may also increase cancer risk in adults because nitrate is endogenously reduced to nitrite and subsequent nitrosation reactions give rise to N-nitroso compounds (NOCs), which are highly carcinogenic and can act systemically.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jansone-Popova, Santa; Ivanov, Alexander S.; Bryantsev, Vyacheslav S.

Here, we report a new family of preorganized bislactam- 1,10-phenanthroline (BLPhen) complexants that possess both hard and soft donor atoms within a convergent cavity and show unprecedented extraction strength for the trivalent fblock metal ions. BLPhen ligands with saturated and unsaturated δ-lactam rings have notable differences in their affinity and selectivity for Am(III) over Eu(III), with the latter being the most selective mixed N,O-donor extractant of Am(III) reported to date. Saturated BLPhen was crystallized with five Ln(III) nitrates to form charge-neutral 1:1 complexes in the solid state. DFT calculations further elaborate on the variety of effects that dictate the performancemore » of these preorganized compounds.« less

Identifying nitrate sources and transformations in surface water by combining dual isotopes of nitrate and stable isotope mixing model in a watershed with different land uses and multi-tributaries

NASA Astrophysics Data System (ADS)

Wang, Meng; Lu, Baohong

2017-04-01

Nitrate is essential for the growth and survival of plants, animals and humans. However, excess nitrate in drinking water is regarded as a health hazard as it is linked to infant methemoglobinemia and esophageal cancer. Revealing nitrate characteristics and identifying its sources are fundamental for making effective water management strategies, but nitrate sources in multi-tributaries and mixed land covered watersheds remain unclear. It is difficult to determine the predominant NO3- sources using conventional water quality monitoring techniques. In our study, based on 20 surface water sampling sites for more than two years' monitoring from April 2012 to December 2014, water chemical and dual isotopic approaches (δ15N-NO3- and δ18O-NO3-) were integrated for the first time to evaluate nitrate characteristics and sources in the Huashan watershed, Jianghuai hilly region, East China. The results demonstrated that nitrate content in surface water was relatively low in the downstream (<10 mg/L), but spatial heterogeneities were remarkable among different sub-watersheds. Extremely high nitrate was observed at the source of the river in one of the sub-watersheds, which exhibited an exponential decline along the stream due to dilution, absorption by aquatic plants, and high forest cover. Although dramatically decline of nitrate occurred along the stream, denitrification was not found in surface water by analyzing δ15N-NO3- and δ18O-NO3- relationship. Proportional contributions of five potential nitrate sources (i.e., precipitation; manure and sewage; soil nitrogen; nitrate fertilizer; nitrate derived from ammonia fertilizer and rainfall) were estimated using a Bayesian isotope mixing model. Model results indicated nitrate sources varied significantly among different rainfall conditions, land use types, as well as anthropologic activities. In summary, coupling dual isotopes of nitrate (δ15N-NO3- and δ18O-NO3-, simultaneously) with a Bayesian isotope mixing model offers a useful and practical way to qualitatively analyze nitrate sources and transformations as well as quantitatively estimate the contributions of potential nitrate sources in surface water. With the assessment of nitrate sources and characteristics, effective management strategies can be implemented to reduce N export and improve water quality in this region.

It is rocket science - why dietary nitrate is hard to 'beet'! Part II: further mechanisms and therapeutic potential of the nitrate-nitrite-NO pathway.

PubMed

Mills, Charlotte Elizabeth; Khatri, Jibran; Maskell, Perry; Odongerel, Chimed; Webb, Andrew James

2017-01-01

Dietary nitrate (found in green leafy vegetables such as rocket and in beetroot) is now recognized to be an important source of nitric oxide, via the nitrate-nitrite-NO pathway. Dietary nitrate confers several cardiovascular beneficial effects on blood pressure, platelets, endothelial function, mitochondrial efficiency and exercise. Having described key twists and turns in the elucidation of the pathway and the underlying mechanisms in Part I, we explore the more recent developments which have served to confirm mechanisms, extend our understanding, and discover new properties and potential therapeutic uses of the pathway in Part II. Even the established dependency on low oxygen states for bioactivation of nitrite has recently been challenged. Dietary nitrate appears to be an important component of 'healthy diets', such as the DASH diet to lower blood pressure and the Mediterranean diet, with its potential to lower cardiovascular risk, possibly through beneficial interactions with a range of other constituents. The World Cancer Research Foundation report strong evidence for vegetables including spinach and lettuce (high nitrate-containing) decreasing cancer risk (mouth, pharynx, larynx, oesophagus and stomach), summarized in a 'Nitrate-Cancer Risk Veg-Table'. The European Space Agency recommends that beetroot, lettuce, spinach and rocket (high-nitrate vegetables) are grown to provide food for long-term space missions. Nitrate, an ancient component of rocket fuel, could support sustainable crops for healthy humans. © 2016 The British Pharmacological Society.

Inter-residue coupling contributes to high-affinity subtype-selective binding of α-bungarotoxin to nicotinic receptors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sine, Steven M.; Huang, Sun; Li, Shu-Xing

2013-09-01

The crystal structure of a pentameric α7 ligand-binding domain chimaera with bound α-btx (α-bungarotoxin) showed that of the five conserved aromatic residues in α7, only Tyr 184 in loop C of the ligand-binding site was required for high-affinity binding. To determine whether the contribution of Tyr 184 depends on local residues, we generated mutations in an α7/5HT 3A (5-hydroxytryptamine type 3A) receptor chimaera, individually and in pairs, and measured 125I-labelled α-btx binding. The results show that mutations of individual residues near Tyr 184 do not affect α-btx affinity, but pairwise mutations decrease affinity in an energetically coupled manner. Kinetic measurementsmore » show that the affinity decreases arise through increases in the α-btx dissociation rate with little change in the association rate. Replacing loop C in α7 with loop C from the α-btx-insensitive α2 or α3 subunits abolishes high-affinity α-btx binding, but preserves acetylcholine-elicited single channel currents. However, in both the α2 and α3 construct, mutating either residue that flanks Tyr 184 to its α7 counterpart restores high-affinity α-btx binding. Analogously, in α7, mutating both residues that flank Tyr 184 to the α2 or α3 counterparts abolishes high-affinity α-btx binding. Thus interaction between Tyr 184 and local residues contributes to high-affinity subtype-selective α-btx binding.« less

Source apportionment of PM2.5 nitrate and sulfate in China using a source-oriented chemical transport model

NASA Astrophysics Data System (ADS)

Zhang, Hongliang; Li, Jingyi; Ying, Qi; Yu, Jian Zhen; Wu, Dui; Cheng, Yuan; He, Kebin; Jiang, Jingkun

2012-12-01

Nitrate and sulfate account for a significant fraction of PM2.5 mass and are generally secondary in nature. Contributions to these two inorganic aerosol components from major sources need to be identified for policy makers to develop cost effective regional emission control strategies. In this work, a source-oriented version of the Community Multiscale Air Quality (CMAQ) model that directly tracks the contributions from multiple emission sources to secondary PM2.5 is developed to determine the regional contributions of power, industry, transportation and residential sectors as well as biogenic sources to nitrate and sulfate concentrations in China in January and August 2009.The source-oriented CMAQ model is capable of reproducing most of the available PM10 and PM2.5 mass, and PM2.5 nitrate and sulfate observations. Model prediction suggests that monthly average PM2.5 inorganic components (nitrate + sulfate + ammonium ion) can be as high as 60 μg m-3 in January and 45 μg m-3 in August, accounting for 20-40% and 50-60% of total PM2.5 mass. The model simulations also indicate significant spatial and temporal variation of the nitrate and sulfate concentrations as well as source contributions in the country. In January, nitrate is high over Central and East China with a maximum of 30 μg m-3 in the Sichuan Basin. In August, nitrate is lower and the maximum concentration of 16 μg m-3 occurs in North China. In January, highest sulfate occurs in the Sichuan Basin with a maximum concentration of 18 μg m-3 while in August high sulfate concentration occurs in North and East China with a similar maximum concentration. Power sector is the dominating source of nitrate and sulfate in both January and August. Transportation sector is an important source of nitrate (20-30%) in both months. Industry sector contributes to both nitrate and sulfate concentrations by approximately 20-30%. Residential sector contributes to approximately 10-20% of nitrate and sulfate in January but its contribution is low in August.

[Interaction of human factor X with thromboplastin].

PubMed

Kiselev, S V; Zubairov, D M; Timarbaev, V N

2003-01-01

The binding of 125I-labeled human factor X to native and papaine-treated tissue tromboplastin in the presence of CaCl2 or EDTA was studied. The Scatchard analysis suggests the existence of high (Kd=l,8 x10(-9) M) and low affinity binding sites on the thromboplastin surface. The removal of Ca2+ reduced affinity of factor X to the high affinity sites. This was accompanied by some increase of their number. Proteolysis by papaine decreased affinity of high affinity sites and caused the increase of their number in the presence of Ca2+. In the absence of Ca2+ the affinity remained unchanged, but the number of sites decreased. At low concentrations of factor X positive cooperativity for high affinity binding sites was observed. It did not depend on the presence of Ca2+. The results indirectly confirm the role of hydrophobic interactons in Ca2+ dependent binding of factor X to thromboplastin and the fact that heterogeneity of this binding is determined by mesophase structure of the thromboplastin phospholipids.

Enhanced Requirement for TNFR2 in Graft Rejection Mediated by Low Affinity Memory CD8+ T Cells During Heterologous Immunity

PubMed Central

Krummey, Scott M.; Chen, Ching-Wen; Guasch, Sara A.; Liu, Danya; Wagener, Maylene; Larsen, Christian P; Ford, Mandy L.

2016-01-01

The affinity of a T cell receptor (TCR) binding to peptide:MHC profoundly impacts the phenotype and function of effector and memory cell differentiation. Little is known about the effect of low affinity priming on memory cell generation and function, which is particularly important in heterologous immunity, when microbe-specific T cells cross-react with allogeneic antigen and mediate graft rejection. We found that low affinity primed memory CD8+ T cells produced high levels of TNF ex vivo in response to heterologous rechallenge compared to high affinity primed memory T cells. Low affinity secondary effectors significantly upregulated TNFR2 on the cell surface and contained a higher frequency of TNFR2hi proliferating cells. Low affinity primed secondary effectors concurrently downregulated TNF production. Importantly, blockade of TNFR2 attenuated graft rejection in low but not high affinity primed animals. These data establish a functional connection between TNF signaling and TCR priming affinity and have implications for the immunomodulation of pathogenic T cell responses during transplantation. PMID:27481849

Estimating Discharge and Nonpoint Source Nitrate Loading to Streams From Three End-Member Pathways Using High-Frequency Water Quality Data

NASA Astrophysics Data System (ADS)

Miller, Matthew P.; Tesoriero, Anthony J.; Hood, Krista; Terziotti, Silvia; Wolock, David M.

2017-12-01

The myriad hydrologic and biogeochemical processes taking place in watersheds occurring across space and time are integrated and reflected in the quantity and quality of water in streams and rivers. Collection of high-frequency water quality data with sensors in surface waters provides new opportunities to disentangle these processes and quantify sources and transport of water and solutes in the coupled groundwater-surface water system. A new approach for separating the streamflow hydrograph into three components was developed and coupled with high-frequency nitrate data to estimate time-variable nitrate loads from chemically dilute quick flow, chemically concentrated quick flow, and slowflow groundwater end-member pathways for periods of up to 2 years in a groundwater-dominated and a quick-flow-dominated stream in central Wisconsin, using only streamflow and in-stream water quality data. The dilute and concentrated quick flow end-members were distinguished using high-frequency specific conductance data. Results indicate that dilute quick flow contributed less than 5% of the nitrate load at both sites, whereas 89 ± 8% of the nitrate load at the groundwater-dominated stream was from slowflow groundwater, and 84 ± 25% of the nitrate load at the quick-flow-dominated stream was from concentrated quick flow. Concentrated quick flow nitrate concentrations varied seasonally at both sites, with peak concentrations in the winter that were 2-3 times greater than minimum concentrations during the growing season. Application of this approach provides an opportunity to assess stream vulnerability to nonpoint source nitrate loading and expected stream responses to current or changing conditions and practices in watersheds.

Synoptic Sampling to Determine Distributed Groundwater-Surface Water Nitrate Loading and Removal Potential Along a Lowland River

NASA Astrophysics Data System (ADS)

Pai, Henry; Villamizar, Sandra R.; Harmon, Thomas C.

2017-11-01

Delineating pollutant reactive transport pathways that connect local land use patterns to surface water is an important goal. This work illustrates high-resolution river mapping of salinity or specific conductance (SC) and nitrate (NO3-) as a potential part of achieving this goal. We observed longitudinal river SC and nitrate distributions using high-resolution synoptic in situ sensing along the lower Merced River (38 river km) in Central California (USA) from 2010 to 2012. We calibrated a distributed groundwater-surface water (GW-SW) discharge model for a conservative solute using 13 synoptic SC sampling events at flows ranging from 1.3 to 31.6 m3 s-1. Nitrogen loads ranged from 0.3 to 1.6 kg N d-1 and were greater following an extended high flow period during a wet winter. Applying the distributed GW-SW discharge estimates to a simplistic reactive nitrate transport model, the model reproduced observed river nitrate distribution well (RRMSE = 5-21%), with dimensionless watershed-averaged nitrate removal (kt) ranging from 0 to 0.43. Estimates were uncertain due to GW nitrate data variability, but the resulting range was consistent with prior removal estimates. At the segment scale, estimated GW-SW nitrate loading ranged from 0 to 17 g NO3- s-1 km-1. Local loading peaked near the middle of the study reach, a location that coincides with a shallow clay lens and with confined animal feed operations in close proximity to the river. Overall, the results demonstrate the potential for high-resolution synoptic monitoring to support GW-SW modeling efforts aimed at understanding and managing nonpoint source pollution.

Thermodynamic Bounds on the Ultra- and Infra-affinity of Hsp70 for Its Substrates

NASA Astrophysics Data System (ADS)

Nguyen, Basile; Hartich, David; Seifert, Udo; Rios, Paolo De Los

2017-07-01

The 70 kDa Heat Shock Proteins Hsp70 have several essential functions in living systems, such as protecting cells against protein aggregation, assisting protein folding, remodeling protein complexes and driving the translocation into organelles. These functions require high affinity for non-specific amino-acid sequences that are ubiquitous in proteins. It has been recently shown that this high affinity, called ultra-affinity, depends on a process driven out of equilibrium by ATP hydrolysis. Here we establish the thermodynamic bounds for ultra-affinity, and further show that the same reaction scheme can in principle be used both to strengthen and to weaken affinities (leading in this case to infra-affinity). We show that cofactors are essential to achieve affinity beyond the equilibrium range. Finally, biological implications are discussed.

Association of nitrogen compounds in drinking water with incidence of esophageal squamous cell carcinoma in Shexian, China.

PubMed

Zhang, Nan; Yu, Cao; Wen, Denggui; Chen, Jun; Ling, Yiwei; Terajima, Kenshi; Akazawa, Kohei; Shan, Baoen; Wang, Shijie

2012-01-01

The incidence of esophageal squamous cell carcinoma (ESCC), which is the eighth most common malignancy worldwide, is highest in China. The purpose of this study was to investigate the association between nitrogen compounds in drinking water with the incidence of ESCC by geographical spatial analysis. The incidence of ESCC is high in Shexian county, China, and environmental factors, particularly nitrogen-contaminated drinking water, are the main suspected risk factors. This study focuses on three nitrogen compounds in drinking water, namely, nitrates, nitrites, and ammonia, all of which are derived mainly from domestic garbage and agricultural fertilizer. The study surveyed 48 villages in the Shexian area with a total population of 54,716 (661 adults with ESCC and 54,055 non-cancer subjects). Hot-spot analysis was used to identify spatial clusters with a high incidence of ESCC and a high concentration of nitrogen compounds. Logistic regression analysis was used to detect risk factors for ESCC incidence. Most areas with high concentrations of nitrate nitrogen in drinking water had a high incidence of ESCC. Correlation analysis revealed a significant positive relationship between nitrate concentration and ESCC (P = 0.01). Logistic regression analysis also confirmed that nitrate nitrogen has a significantly higher odds ratio. The results indicate that nitrate nitrogen is associated with ESCC incidence in Shexian county. In conclusion, high concentrations of nitrate nitrogen in drinking water may be a significant risk factor for the incidence of ESCC.

Nitrate removal in deep sediments of a nitrogen-rich river network: A test of a conceptual model

USGS Publications Warehouse

Stelzer, Robert S.; Bartsch, Lynn

2012-01-01

Many estimates of nitrogen removal in streams and watersheds do not include or account for nitrate removal in deep sediments, particularly in gaining streams. We developed and tested a conceptual model for nitrate removal in deep sediments in a nitrogen-rich river network. The model predicts that oxic, nitrate-rich groundwater will become depleted in nitrate as groundwater upwelling through sediments encounters a zone that contains buried particulate organic carbon, which promotes redox conditions favorable for nitrate removal. We tested the model at eight sites in upwelling reaches of lotic ecosystems in the Waupaca River Watershed that varied by three orders of magnitude in groundwater nitrate concentration. We measured denitrification potential in sediment core sections to 30 cm and developed vertical nitrate profiles to a depth of about 1 m with peepers and piezometer nests. Denitrification potential was higher, on average, in shallower core sections. However, core sections deeper than 5 cm accounted for 70%, on average, of the depth-integrated denitrification potential. Denitrification potential increased linearly with groundwater nitrate concentration up to 2 mg NO3-N/L but the relationship broke down at higher concentrations (> 5 mg NO3-N/L), a pattern that suggests nitrate saturation. At most sites groundwater nitrate declined from high concentrations at depth to much lower concentrations prior to discharge into the surface water. The profiles suggested that nitrate removal occurred at sediment depths between 20 and 40 cm. Dissolved oxygen concentrations were much higher in deep sediments than in pore water at 5 cm sediment depth at most locations. The substantial denitrification potential in deep sediments coupled with the declines in nitrate and dissolved oxygen concentrations in upwelling groundwater suggest that our conceptual model for nitrate removal in deep sediments is applicable to this river network. Our results suggest that nitrate removal rates can be high in deep sediments of upwelling stream reaches, which may have implications for efforts to understand and quantify nitrogen transport and removal at larger scales.

Can nitrate contaminated groundwater be remediated by optimizing flood irrigation rate with high nitrate water in a desert oasis using the WHCNS model?

PubMed

Liang, Hao; Qi, Zhiming; Hu, Kelin; Prasher, Shiv O; Zhang, Yuanpei

2016-10-01

Nitrate contamination of groundwater is an environmental concern in intensively cultivated desert oases where this polluted groundwater is in turn used as a major irrigation water resource. However, nitrate fluxes from root zone to groundwater are difficult to monitor in this complex system. The objectives of this study were to validate and apply the WHCNS (soil Water Heat Carbon Nitrogen Simulator) model to simulate water drainage and nitrate leaching under different irrigation and nitrogen (N) management practices, and to assess the utilization of groundwater nitrate as an approach to remediate nitrate contaminated groundwater while maintain crop yield. A two-year field experiment was conducted in a corn field irrigated with high nitrate groundwater (20 mg N L(-1)) in Alxa, Inner Mongolia, China. The experiment consisted of two irrigation treatments (Istd, standard, 750 mm per season; Icsv, conservation, 570 mm per season) factorially combined with two N fertilization treatments (Nstd, standard, 138 kg ha(-1); Ncsv, conservation, 92 kg ha(-1)). The validated results showed that the WHCNS model simulated values of crop dry matter, yield, soil water content and soil N concentration in soil profile all agreed well with the observed values. Compared to the standard water management (Istd), the simulated drainage and nitrate leaching decreased about 65% and 59%, respectively, under the conservation water management (Icsv). Nearly 55% of input N was lost by leaching under the IstdNstd and IstdNcsv treatments, compared to only 26% under the IcsvNstd and IcsvNcsv treatments. Simulations with more than 240 scenarios combing different levels of irrigation and fertilization indicated that irrigation was the main reason leading to the high risk of nitrate leaching, and the nitrate in irrigation groundwater can be best utilized without corn yield loss when the total irrigation was reduced from the current 750 mm to 491 mm. This reduced irrigation rate facilitated the use of approximately 42 kg N ha(-1) yr(-1) of nitrate from groundwater, which would gradually improve the groundwater quality. Future field studies on nitrate leaching in this area are suggested to investigate water and N dynamics under irrigation rates near 490 mm per season. Copyright © 2016 Elsevier Ltd. All rights reserved.

Dynamic modeling of nitrogen losses in river networks unravels the coupled effects of hydrological and biogeochemical processes

USGS Publications Warehouse

Alexander, Richard B.; Böhlke, John Karl; Boyer, Elizabeth W.; David, Mark B.; Harvey, Judson W.; Mulholland, Patrick J.; Seitzinger, Sybil P.; Tobias, Craig R.; Tonitto, Christina; Wollheim, Wilfred M.

2009-01-01

The importance of lotic systems as sinks for nitrogen inputs is well recognized. A fraction of nitrogen in streamflow is removed to the atmosphere via denitrification with the remainder exported in streamflow as nitrogen loads. At the watershed scale, there is a keen interest in understanding the factors that control the fate of nitrogen throughout the stream channel network, with particular attention to the processes that deliver large nitrogen loads to sensitive coastal ecosystems. We use a dynamic stream transport model to assess biogeochemical (nitrate loadings, concentration, temperature) and hydrological (discharge, depth, velocity) effects on reach-scale denitrification and nitrate removal in the river networks of two watersheds having widely differing levels of nitrate enrichment but nearly identical discharges. Stream denitrification is estimated by regression as a nonlinear function of nitrate concentration, streamflow, and temperature, using more than 300 published measurements from a variety of US streams. These relations are used in the stream transport model to characterize nitrate dynamics related to denitrification at a monthly time scale in the stream reaches of the two watersheds. Results indicate that the nitrate removal efficiency of streams, as measured by the percentage of the stream nitrate flux removed via denitrification per unit length of channel, is appreciably reduced during months with high discharge and nitrate flux and increases during months of low-discharge and flux. Biogeochemical factors, including land use, nitrate inputs, and stream concentrations, are a major control on reach-scale denitrification, evidenced by the disproportionately lower nitrate removal efficiency in streams of the highly nitrate-enriched watershed as compared with that in similarly sized streams in the less nitrate-enriched watershed. Sensitivity analyses reveal that these important biogeochemical factors and physical hydrological factors contribute nearly equally to seasonal and stream-size related variations in the percentage of the stream nitrate flux removed in each watershed.

Effects of agricultural practices and vadose zone stratigraphy on nitrate concentration in ground water in Kansas, USA

USGS Publications Warehouse

Townsend, M.A.; Sleezer, R.O.; Macko, S.A.; ,

1996-01-01

Differences in nitrate-N concentrations in,around water in Kansas can be explained by variations in agricultural practices and vadose-zone stratigraphy. In northwestern Kansas, past use of a local stream for tailwater runoff from irrigation and high fertilizer applications for sugar-beet farming resulted in high nitrate-N concentrations (12-60 mg L-1; in both soil and ground water. Nitrogen isotope values from the soil and ground water range from +4 to +8? which is typical for a fertilizer source. In parts of south-central Kansas, the use of crop rotation and the presence of both continuous fine-textured layers and a reducing ground-water chemistry resulted in ground-water nitrate-N values of 10 mg L-1; in both soil and grounwater. Nitrogen isotope values of +3 to +7? indicate a fertilizer source. Crop rotation decreased nitrate-N values in the shallow ground water (9 m). However, deeper ground water showed increasing nitrate-N concentrations as a result of past farming practices.

High-density defects on PdAg nanowire networks as catalytic hot spots for efficient dehydrogenation of formic acid and reduction of nitrate.

PubMed

Liu, Hu; Yu, Yongsheng; Yang, Weiwei; Lei, Wenjuan; Gao, Manyi; Guo, Shaojun

2017-07-13

Controlling the surface defects of nanocrystals is a new way of tuning/boosting their catalytic properties. Herein, we report networked PdAg nanowires (NWs) with high-density defects as catalytic hot spots for efficient catalytic dehydrogenation of formic acid (FA) and catalytic reduction of nitrates. The networked PdAg NWs exhibit composition-dependent catalytic activity for the dehydrogenation reaction of FA without any additive, with Pd 5 Ag 5 NWs exhibiting the highest activity. They also show good durability, reflected by the retention of their initial activity during the dehydrogenation reaction of FA even after five cycles. Their initial TOF is 419 h -1 at 60 °C in water solution, much higher than those of the most Pd-based catalysts with a support. Moreover, they can efficiently reduce nitrates to alleviate nitrate pollution in water (conversion yield >99%). This strategy opens up a new green synthetic technique to design support-free heterogeneous catalysts with high-density defects as catalytic hot spots for efficient dehydrogenation catalysis of FA to meet the requirement of fuel cell applications and catalytic reduction of nitrates in water polluted with nitrates.

The formation of nitrogen-containing organic oxidation products in a heavily polluted urban environment

NASA Astrophysics Data System (ADS)

Chen, Q.; Cheng, X.; Zheng, Y.; Li, Y.; Zhu, T.; Zhang, Q.; Canagaratna, M. R.; Nowark, J.; Worsnop, D. R.

2017-12-01

Nitrogen-containing organic oxidation products are important species that may contribute to secondary organic aerosol and to redistribute nitrogen oxides through photolysis and oxidation. We deployed a nitrate ion chemical ionization time-of-flight mass spectrometer and a long time-of-flight aerosol mass spectrometer in Beijing for various seasons to study the nitrogen-containing organic species in both gas and particle phases. High concentrations of nitrated phenols were observed in both winter and summer, due to contributions from both primary and secondary sources. The concentrations of gaseous dinitrophenols tracked the severe haze events and correlated well with many highly oxygenated organic molecules. We also identified mass spectral tracers for quantifying organic nitrates. Significant photochemical production of nitrate and organic nitrates were evident during severe haze events, which may contribute to persistent particle formation. The findings are consistent with observed high OH turnover rates, highlighting the importance of gaseous oxidation pathways on persistent particle formation during haze.

A study of the uptake of chloroquine in malaria-infected erythrocytes. High and low affinity uptake and the influence of glucose and its analogues.

PubMed

Diribe, C O; Warhurst, D C

1985-09-01

A study of concentration- and substrate-dependence of chloroquine uptake has been carried out on mouse erythrocytes infected with the chloroquine-sensitive NK65 and the chloroquine-resistant RC strains of Plasmodium berghei. The presence of drug binding sites of high and low affinity in such strains of P. berghei was confirmed. High affinity uptake sites in cells parasitized with chloroquine-sensitive and chloroquine-resistant parasites have similar characteristics, but in the sensitive strain the major component of chloroquine-uptake is at high affinity and dependent on the availability of ATP whilst in the resistant strain the major component of uptake is at low affinity and independent of energy. An absolute increase in the quantity of the low affinity site in erythrocytes parasitized with chloroquine-resistant P. berghei was noted, which may be related to an increase in quantity of parasite membrane.

Persistent Influences of the 2002 Hayman Fire on Stream Nitrate and Dissolved Organic Carbon

NASA Astrophysics Data System (ADS)

Rhoades, C.; Pierson, D. N.; Fegel, T. S., II; Chow, A. T.; Covino, T. P.

2016-12-01

Large, high severity wildfires alter the physical and biological conditions that determine how watersheds retain and release nutrients and regulate stream water quality. For five years after the 2002 Hayman Fire burned in Colorado conifer forests, stream nitrate concentrations and export increased steadily in watersheds with extensive high-severity burning. Stream temperature and turbidity also increased in relation to the extent of high-severity burning and remained elevated above background levels throughout the initial five year post-fire period. Our recent sampling documents that 14 years after the Hayman Fire stream nitrate remains an order of magnitude higher in extensively-burned (35-90%) compared to unburned watersheds (0.2 vs 2.8 mg L-1). Nitrate represents 83% of the total dissolved N in extensively-burned watersheds compared to 29% in unburned watersheds. In contrast, dissolved organic carbon (DOC), was highest in watersheds that burned to a moderate extent (10-20%) and lowest in those with extensive burning. Catchments with a moderate extent burned had DOC concentrations 2.5 and 1.7 times more than those with extensive burning and unburned catchments, respectively. Peak concentrations of DOC and nitrate track the rising limb of the streamflow hydrograph and reach a maximum in May, but patterns among burn extent categories were seasonally consistent. Current riparian conditions are linked to stream nitrate in burned watersheds. For example, stream nitrate increases proportionally to the extent of riparian zones with low shrub cover (R2 = 0.76). We found signs of watershed recovery compared to the initial post-fire period; stream temperature and turbidity remained elevated in extensively burned catchments, but increases were only significant during the spring season. The persistent stream nitrate concentrations as well as the relation between riparian cover and post-fire stream nitrate may help prioritize restoration planting efforts and mitigate chronic, elevated nitrate export from burned watersheds.

Time-trial Performance in World-Class Speed Skaters After Chronic Nitrate Ingestion.

PubMed

Richard, Philippe; Koziris, Lymperis P; Charbonneau, Mathieu; Naulleau, Catherine; Tremblay, Jonathan; Billaut, François

2018-05-10

Nitrate supplementation can increase tolerance to high-intensity work rates, however limited data exist on the recovery of performance. We tested whether 5 days of nitrate supplementation could improve repeated time-trial performance in speed skating. Using a double-blind, placebo-controlled, crossover design, 9 international-level short-track speed skaters ingested one high (juice blend, ~6.5mmol nitrate; HI) or low dose (juice blend, ~1mmol nitrate; LO) per day on days 1-4. After a double dose of either HI or LO on day 5, athletes performed two on-ice 1000-m time trials, separated by 35 min, to simulate competition races. Differences between HI and LO were compared to the smallest practically important difference. Salivary [nitrate] and [nitrite] were higher in HI than LO before the first (nitrate 81%, ES 1.76; nitrite 72%, ES 1.73) and second pursuits (nitrate 81%, ES 1.92; nitrite 71%, ES 1.78). However, there was no difference in performance in the first (LO: 90.92±4.08sec; HI: 90.95±4.06sec, ES 0.01) nor the second time-trial (LO: 91.16±4.06; HI: 91.55±4.40sec, ES 0.09). Plasma [lactate] measured after the trials (LO: 14.8±1.1mM; HI: 14.8±1.2mM, ES 0.01) and at the end of the recovery period (LO: 9.8±2.1mM; HI: 10.2±1.9mM, ES 0.05) was not different between treatments. Five days of a high-dose nitrate supplementation did not change physiological responses and failed to improve single and repeated time-trial performances in world-class short-track speed skaters. These data suggest that nitrate ingestion up to 6.5mmol does not enhance recovery from supra-maximal exercise in world-class athletes.

Tracking nitrate sources in the Chaohu Lake, China, using the nitrogen and oxygen isotopic approach.

PubMed

Yu, Qibiao; Wang, Fang; Li, Xinyan; Yan, Weijin; Li, Yanqiang; Lv, Shucong

2018-05-07

The Chaohu Lake is highly polluted and suffers from severe eutrophication. Nitrate is a key form of nitrogen that can cause water quality degradation. In this study, hydrochemical and dual isotopic approaches were utilized to identify the seasonal variation of nitrate sources in the Chaohu Lake and its inflowing rivers. The average nitrate concentrations ([NO 3 - ]) of the lake and its inflowing rivers were 89.9 and 140.8 μmol L -1 , respectively. The isotopic values of δ 15 N-NO 3 - and δ 18 O-NO 3 - in the lake ranged from - 1.01 to + 16.67‰ and from - 4.39 to + 22.20‰, respectively. The two major rivers had distinct isotopic compositions, with average δ 15 N-NO 3 - values of + 17.52 and + 3.51‰, and average δ 18 O-NO 3 - values of + 2.71 and + 7.47‰ for the Nanfei River and Hangbu River, respectively. The results show that soil organic ammonium and urban wastewater discharge were the main sources of nitrate in the Chaohu Lake, and nitrate assimilation was an important process affected [NO 3 - ] and isotopic compositions, especially in the western Chaohu Lake. The elevated [NO 3 - ] and δ 15 N-NO 3 - values in the western Chaohu Lake indicate the high influence of human activities. Urban wastewater discharge was the primary nitrate source in the Nanfei River and soil organic ammonium was the main source in the Hangbu River. Although nitrate from direct atmospheric deposition was low, its strong flushing effect can substantially improve riverine [NO 3 - ] and nitrate loading from terrestrial ecosystem. The relatively high nitrate fluxes from the Hangbu River indicate that nitrogen loading from agricultural watershed is unneglectable in watershed nitrogen sources reduction strategies.

Nitrate Intake Does Not Influence Bladder Cancer Risk: The Netherlands Cohort Study

PubMed Central

Zeegers, Maurice P.; Selen, Roel F.M.; Kleinjans, Jos C.S.; Goldbohm, R. Alexandra; van den Brandt, Piet A.

2006-01-01

Objectives N-nitroso compounds, endogenously formed from nitrate-derived nitrite, are suspected to be important bladder carcinogens. However, the association between nitrate exposure from food or drinking water and bladder cancer has not been substantially investigated in epidemiologic studies. Methods We evaluated the associations between nitrate exposure and bladder cancer in the Netherlands Cohort Study, conducted among 120,852 men and women, 55–69 years of age at entry. Information on nitrate from diet was collected via a food frequency questionnaire in 1986 and a database on nitrate content of foods. Individual nitrate exposures from beverages prepared with tap water were calculated by linking the postal code of individual residence at baseline to water company data. After 9.3 years of follow-up and after excluding subjects with incomplete or inconsistent dietary data, 889 cases and 4,441 subcohort members were available for multivariate analyses. We calculated incidence rate ratios (RR) and corresponding 95% confidence intervals (CIs) using Cox regression analyses. We also evaluated possible effect modification of dietary intake of vitamins C and E (low/high) and cigarette smoking (never/ever). Results The multivariate RRs for nitrate exposure from food, drinking water, and estimated total nitrate exposure were 1.06 (95% CI, 0.81–1.31), 1.06 (95% CI, 0.82–1.37), and 1.09 (95% CI, 0.84–1.42), respectively, comparing the highest to the lowest quintiles of intake. Dietary intake of vitamins C and E (low/high) and cigarette smoking (never/ever) had no significant impact on these results. Conclusion Although the association between nitrate exposure and bladder cancer risk is biologically plausible, our results in this study do not support an association between nitrate exposure and bladder cancer risk. PMID:17035137

Effect of Ammonium and Nitrate on Ferric Chelate Reductase and Nitrate Reductase in Vaccinium Species

PubMed Central

POONNACHIT, U.; DARNELL, R.

2004-01-01

• Background and Aims Most Vaccinium species have strict soil requirements for optimal growth, requiring low pH, high iron availability and nitrogen primarily in the ammonium form. These soils are limited and are often located near wetlands. Vaccinium arboreum is a wild species adapted to a wide range of soils, including high pH, low iron, and nitrate‐containing soils. This broader soil adaptation in V. arboreum may be related to increased efficiency of iron or nitrate uptake compared with the cultivated Vaccinium species. • Methods Nitrate, ammonium and iron uptake, and nitrate reductase (NR) and ferric chelate reductase (FCR) activities were compared in two Vaccinium species grown hydroponically in either nitrate or ammonia, with or without iron. The species studied were the wild V. arboreum and the cultivated V. corymbosum interspecific hybrid, which exhibits the strict soil requirements of most Vaccinium species. • Key Results Ammonium uptake was significantly greater than nitrate uptake in both species, while nitrate uptake was greater in the wild species, V. arboreum, compared with the cultivated species, V. corymbosum. The increased nitrate uptake in V. arboreum was correlated with increased root NR activity compared with V. corymbosum. The lower nitrate uptake in V. corymbosum was reflected in decreased plant dry weight in this species compared with V. arboreum. Root FCR activity increased significantly in V. corymbosum grown under iron‐deficient conditions, compared with the same species grown under iron‐sufficient conditions or with V. arboreum grown under either iron condition. • Conclusions. V. arboreum appears to be more efficient in acquiring nitrate compared with V. corymbosum, possibly due to increased NR activity and this may partially explain the wider soil adaptation of V. arboreum. PMID:14980973

Number of nitrate groups determines reactivity and potency of organic nitrates: a proof of concept study in ALDH-2−/− mice

PubMed Central

Wenzel, P; Hink, U; Oelze, M; Seeling, A; Isse, T; Bruns, K; Steinhoff, L; Brandt, M; Kleschyov, A L; Schulz, E; Lange, K; Weiner, H; Lehmann, J; Lackner, K J; Kawamoto, T; Münzel, T; Daiber, A

2007-01-01

Background and purpose: Mitochondrial aldehyde dehydrogenase (ALDH-2) has been shown to provide a pathway for bioactivation of organic nitrates and to be prone to desensitization in response to highly potent, but not to less potent, nitrates. We therefore sought to support the hypothesis that bioactivation by ALDH-2 critically depends on the number of nitrate groups within the nitrovasodilator. Experimental approach: Nitrates with one (PEMN), two (PEDN; GDN), three (PETriN; glyceryl trinitrate, GTN) and four (pentaerithrityl tetranitrate, PETN) nitrate groups were investigated. Vasodilatory potency was measured in isometric tension studies using isolated aortic segments of wild type (WT) and ALDH-2−/− mice. Activity of the cGMP-dependent kinase-I (reflected by levels of phosphorylated VAsodilator Stimulated Phosphoprotein, P-VASP) was quantified by Western blot analysis, mitochondrial dehydrogenase activity by HPLC. Following incubation of isolated mitochondria with PETN, PETriN-chromophore and PEDN, metabolites were quantified using chemiluminescence nitrogen detection and mass spectrometry. Key results: Compared to WT, vasorelaxation in response to PETN, PETriN and GTN was attenuated about 10fold in ALDH-2−/− mice, identical to WT vessels preincubated with inhibitors of ALDH-2. Reduced vasodilator potency correlated with reduced P-VASP formation and diminished biotransformation of the tetranitrate- and trinitrate-compounds. None of these findings were observed for PEDN, GDN and PEMN. Conclusions and implications: Our results support the crucial role of ALDH-2 in bioactivating highly reactive nitrates like GTN, PETN and PETriN. ALDH-2-mediated relaxation by organic nitrates therefore depends mainly on the number of nitrate groups. Less potent nitrates like PEDN, GDN and PEMN are apparently biotransformed by other pathways. PMID:17220910

Response of humic acid formation to elevated nitrate during chicken manure composting.

PubMed

Shi, Mingzi; Wei, Zimin; Wang, Liqin; Wu, Junqiu; Zhang, Duoying; Wei, Dan; Tang, Yu; Zhao, Yue

2018-06-01

Nitrate can stimulate microbes to degrade aromatic compounds, whereas humic acid (HA) as a high molecular weight aromatic compound, its formation may be affected by elevated nitrate during composting. Therefore, this study is conducted to determine the effect of elevated nitrate on HA formation. Five tests were executed by adding different nitrate concentrations to chicken manure composting. Results demonstrate that the concentration of HA in treatment group is significantly decreased compared with control group (p < 0.05), especially in the highest nitrate concentration group. RDA indicates that the microbes associated with HA and environmental parameters are influenced by elevated nitrate. Furthermore, structural equation model reveals that elevated nitrate reduces HA formation by mediating microbes directly, or by affecting ammonia and pH as the indirect drivers to regulate microbial community structure. Copyright © 2018 Elsevier Ltd. All rights reserved.

Structural basis for dynamic mechanism of nitrate/nitrite antiport by NarK

NASA Astrophysics Data System (ADS)

Fukuda, Masahiro; Takeda, Hironori; Kato, Hideaki E.; Doki, Shintaro; Ito, Koichi; Maturana, Andrés D.; Ishitani, Ryuichiro; Nureki, Osamu

2015-05-01

NarK belongs to the nitrate/nitrite porter (NNP) family in the major facilitator superfamily (MFS) and plays a central role in nitrate uptake across the membrane in diverse organisms, including archaea, bacteria, fungi and plants. Although previous studies provided insight into the overall structure and the substrate recognition of NarK, its molecular mechanism, including the driving force for nitrate transport, remained elusive. Here we demonstrate that NarK is a nitrate/nitrite antiporter, using an in vitro reconstituted system. Furthermore, we present the high-resolution crystal structures of NarK from Escherichia coli in the nitrate-bound occluded, nitrate-bound inward-open and apo inward-open states. The integrated structural, functional and computational analyses reveal the nitrate/nitrite antiport mechanism of NarK, in which substrate recognition is coupled to the transport cycle by the concomitant movement of the transmembrane helices and the key tyrosine and arginine residues in the substrate-binding site.

Effect of dietary nitrate level on enteric methane production, hydrogen emission, rumen fermentation, and nutrient digestibility in dairy cows.

PubMed

Olijhoek, D W; Hellwing, A L F; Brask, M; Weisbjerg, M R; Højberg, O; Larsen, M K; Dijkstra, J; Erlandsen, E J; Lund, P

2016-08-01

Nitrate may lower methane production in ruminants by competing with methanogenesis for available hydrogen in the rumen. This study evaluated the effect of 4 levels of dietary nitrate addition on enteric methane production, hydrogen emission, feed intake, rumen fermentation, nutrient digestibility, microbial protein synthesis, and blood methemoglobin. In a 4×4 Latin square design 4 lactating Danish Holstein dairy cows fitted with rumen, duodenal, and ileal cannulas were assigned to 4 calcium ammonium nitrate addition levels: control, low, medium, and high [0, 5.3, 13.6, and 21.1g of nitrate/kg of dry matter (DM), respectively]. Diets were made isonitrogenous by replacing urea. Cows were fed ad libitum and, after a 6-d period of gradual introduction of nitrate, adapted to the corn-silage-based total mixed ration (forage:concentrate ratio 50:50 on DM basis) for 16d before sampling. Digesta content from duodenum, ileum, and feces, and rumen liquid were collected, after which methane production and hydrogen emissions were measured in respiration chambers. Methane production [L/kg of dry matter intake (DMI)] linearly decreased with increasing nitrate concentrations compared with the control, corresponding to a reduction of 6, 13, and 23% for the low, medium, and high diets, respectively. Methane production was lowered with apparent efficiencies (measured methane reduction relative to potential methane reduction) of 82.3, 71.9, and 79.4% for the low, medium, and high diets, respectively. Addition of nitrate increased hydrogen emissions (L/kg of DMI) quadratically by a factor of 2.5, 3.4, and 3.0 (as L/kg of DMI) for the low, medium, and high diets, respectively, compared with the control. Blood methemoglobin levels and nitrate concentrations in milk and urine increased with increasing nitrate intake, but did not constitute a threat for animal health and human food safety. Microbial crude protein synthesis and efficiency were unaffected. Total volatile fatty acid concentration and molar proportions of acetate, butyrate, and propionate were unaffected, whereas molar proportions of formate increased. Milk yield, milk composition, DMI and digestibility of DM, organic matter, crude protein, and neutral detergent fiber in rumen, small intestine, hindgut, and total tract were unaffected by addition of nitrate. In conclusion, nitrate lowered methane production linearly with minor effects on rumen fermentation and no effects on nutrient digestibility. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Reactive transport model for bioremediation of nitrate using fumarate in groundwater system: verification and field application

NASA Astrophysics Data System (ADS)

Lee, S.; Yeo, I. W.; Yeum, Y.; Kim, Y.

2016-12-01

Previous studies showed that groundwater of rural areas in Korea is often contaminated with nitrate highly exceeding the drinking water standard of 10 mg/L (NO3-N), which poses a major threat in human and livestock health. In-situ bioremediation method has been developed to reduce high nitrate-nitrogen concentration in groundwater using slowly released encapsulated carbon source. Collaborative research of this study revealed that fumarate was found to be a very effective carbon source in terms of cost and nitrate reduction against formate, propionate, and lactate. For reactive transport modeling of the bioremediation of nitrate using fumarate, the BTEX module of RT3D incorporated in GMS, a commercial groundwater modeling software developed by AQUAVEO, was adopted, where BTEX was replaced with fumarate as a carbon source. Column tests were carried out to determine transport and reaction parameters for numerical modeling such as dispersity and first order degradation rate of nitrate by fumarate. The calibration of the numerical model against column tests strongly indicated that nitrate, known to be not reactive in groundwater system, appeared to be retarded due to sorption by fumarate. The calibrated model was tested for field-scale application to the composting facility in Gimje, Korea. The numerical results showed that the model could simulate the nitrate reduction by fumarate in field scale groundwater system. The reactive transport model for nitrate can be used as a tool for optimum design of in-situ nitrate bioremediation system, such as released depth and amount of fumarate and the spacing of wells that encapsulated fumarate is released through.

Nitrate reductase activity of Staphylococcus carnosus affecting the color formation in cured raw ham.

PubMed

Bosse Née Danz, Ramona; Gibis, Monika; Schmidt, Herbert; Weiss, Jochen

2016-07-01

The influence of the nitrate reductase activity of two Staphylococcus carnosus strains used as starter cultures on the formation of nitrate, nitrite and color pigments in cured raw ham was investigated. In this context, microbiological, chemical and multivariate image analyses were carried out on cured raw hams, which were injected with different brines containing either nitrite or nitrate, with or without the S. carnosus starter cultures. During processing and storage, the viable counts of staphylococci remained constant at 6.5logcfu/g in the hams inoculated with starter cultures, while the background microbiota of the hams processed without the starter cultures developed after 14days. Those cured hams inoculated with S. carnosus LTH 7036 (high nitrate reductase activity) showed the highest decrease in nitrate and high nitrite concentrations in the end product, but were still in the range of the legal European level. The hams cured with nitrate and without starter culture or with the other strain, S. carnosus LTH 3838 (low nitrate reductase activity) showed higher residual nitrate levels and a lower nitrite content in the end product. The multivariate image analysis identified spatial and temporal differences in the meat pigment profiles of the differently cured hams. The cured hams inoculated with S. carnosus LTH 3838 showed an uncured core due to a delay in pigment formation. Therefore, the selection of starter cultures based on their nitrate reductase activity is a key point in the formation of curing compounds and color pigments in cured raw ham manufacture. Copyright © 2016 Elsevier Ltd. All rights reserved.

Thyroid Cancer Induction: Nitrates as Independent Risk Factors or Risk Modulators after Radiation Exposure, with a Focus on the Chernobyl Accident.

PubMed

Drozd, Valentina M; Branovan, Igor; Shiglik, Nikolay; Biko, Johannes; Reiners, Christoph

2018-03-01

In recent decades, differentiated thyroid cancer (DTC) incidence has been increasing worldwide. The important contributions to this phenomenon of "overdiagnosis" driven by wider use of improved ultrasound systems are amply documented, notwithstanding the "real" carcinogenic effects of ionizing radiation, e.g., from the Chernobyl accident or health care interventions. Less well understood is the role of nitrates - as environmental pollutants, in diet, and in medication - in thyroid carcinogenesis. Increasing exposure to nitrates is associated with rising incidence of esophageal, stomach, bladder, and colon cancers. Recent data suggest that in agricultural areas with higher mean nitrate levels in groundwater, DTC risk is also elevated. Our work in Belarus after Chernobyl has shown that children in districts with high nitrate concentrations in drinking water had significantly higher thyroid cancer incidence after irradiation than did their counterparts in areas with lower nitrate concentrations. Notwithstanding thyroid shielding, increasing use of computed tomography and dental X-rays heightens radiation exposure of the salivary glands in the general population, especially in children and adolescents. When nitrate intake is increased, salivary gland irradiation may potentially result in carcinogenic elevations in plasma nitric oxide concentrations. In conclusion, excess nitrate intake seems to be an independent risk factor for DTC. Additionally, we hypothesize from our data that high nitrate levels modulate the carcinogenic effect of radiation on the thyroid. Cohort studies, case-control studies, or both, are needed to quantify the effects of nitrates on DTC risk in the presence or absence of radiation exposure, e.g., that associated with diagnostic or therapeutic health care interventions.

Denitrification using a monopolar electrocoagulation/flotation (ECF) process.

PubMed

Emamjomeh, Mohammad M; Sivakumar, Muttucumaru

2009-01-01

Nitrate levels are limited due to health concerns in potable water. Nitrate is a common contaminant in water supplies, and especially prevalent in surface water supplies and shallow wells. Nitrate is a stable and highly soluble ion with low potential for precipitation or adsorption. These properties make it difficult to remove using conventional water treatment methods. A laboratory batch electrocoagulation/flotation (ECF) reactor was designed to investigate the effects of different parameters such as electrolysis time, electrolyte pH, initial nitrate concentration, and current rate on the nitrate removal efficiency. The optimum nitrate removal was observed at a pH range of between 9 and 11. It appeared that the nitrate removal rate was 93% when the initial nitrate concentration and electrolysis time respectively were 100 mg L(-1)-NO(3)(-) and 40 min. The results showed a linear relationship between the electrolysis time for total nitrate removal and the initial nitrate concentration. It is concluded that the electrocoagulation technology for denitrification can be an effective preliminary process when the ammonia byproduct must be effectively removed by the treatment facilities.

Mass spectrometric identification of intermediates in the O2-driven [4Fe-4S] to [2Fe-2S] cluster conversion in FNR

PubMed Central

Crack, Jason C.; Thomson, Andrew J.

2017-01-01

The iron-sulfur cluster containing protein Fumarate and Nitrate Reduction (FNR) is the master regulator for the switch between anaerobic and aerobic respiration in Escherichia coli and many other bacteria. The [4Fe-4S] cluster functions as the sensory module, undergoing reaction with O2 that leads to conversion to a [2Fe-2S] form with loss of high-affinity DNA binding. Here, we report studies of the FNR cluster conversion reaction using time-resolved electrospray ionization mass spectrometry. The data provide insight into the reaction, permitting the detection of cluster conversion intermediates and products, including a [3Fe-3S] cluster and persulfide-coordinated [2Fe-2S] clusters [[2Fe-2S](S)n, where n = 1 or 2]. Analysis of kinetic data revealed a branched mechanism in which cluster sulfide oxidation occurs in parallel with cluster conversion and not as a subsequent, secondary reaction to generate [2Fe-2S](S)n species. This methodology shows great potential for broad application to studies of protein cofactor–small molecule interactions. PMID:28373574

High-frequency DOC and nitrate measurements provide new insights into their export and their relationships to rainfall-runoff processes

NASA Astrophysics Data System (ADS)

Schwab, Michael; Klaus, Julian; Pfister, Laurent; Weiler, Markus

2015-04-01

Over the past decades, stream sampling protocols for environmental tracers were often limited by logistical and technological constraints. Long-term sampling programs would typically rely on weekly sampling campaigns, while high-frequency sampling would remain restricted to a few days or hours at best. We stipulate that the currently predominant sampling protocols are too coarse to capture and understand the full amplitude of rainfall-runoff processes and its relation to water quality fluctuations. Weekly sampling protocols are not suited to get insights into the hydrological system during high flow conditions. Likewise, high frequency measurements of a few isolated events do not allow grasping inter-event variability in contributions and processes. Our working hypothesis is based on the potential of a new generation of field-deployable instruments for measuring environmental tracers at high temporal frequencies over an extended period. With this new generation of instruments we expect to gain new insights into rainfall-runoff dynamics, both at intra- and inter-event scales. Here, we present the results of one year of DOC and nitrate measurements with the field deployable UV-Vis spectrometer spectro::lyser (scan Messtechnik GmbH). The instrument measures the absorption spectrum from 220 to 720 nm in situ and at high frequencies and derives DOC and nitrate concentrations. The measurements were carried out at 15 minutes intervals in the Weierbach catchment (0.47 km2) in Luxemburg. This fully forested catchment is characterized by cambisol soils and fractured schist as underlying bedrock. The time series of DOC and nitrate give insights into the high frequency dynamics of stream water. Peaks in DOC concentrations are closely linked to discharge peaks that occur during or right after a rainfall event. Those first discharge peaks can be linked to fast near surface runoff processes and are responsible for a remarkable amount of DOC export. A special characterisation of the Weierbach catchment are the delayed second peaks a few days after the rainfall event. Nitrate concentrations are following this second peak. We assume that this delayed response is going back to subsurface or upper groundwater flows, with nitrate enriched water. On an inter-event scale during low flow / base flow conditions, we observe interesting diurnal patterns of both DOC and nitrate concentrations. Overall, the long-term high-frequency measurements of DOC and nitrate provide us the opportunity to separate different rainfall-runoff processes and link the amount of DOC and nitrate export to them to quantify the overall relevance of the different processes.

The role of CH/π interactions in the high affinity binding of streptavidin and biotin.

PubMed

Ozawa, Motoyasu; Ozawa, Tomonaga; Nishio, Motohiro; Ueda, Kazuyoshi

2017-08-01

The streptavidin-biotin complex has an extraordinarily high affinity (Ka: 10 15 mol -1 ) and contains one of the strongest non-covalent interactions known. This strong interaction is widely used in biological tools, including for affinity tags, detection, and immobilization of proteins. Although hydrogen bond networks and hydrophobic interactions have been proposed to explain this high affinity, the reasons for it remain poorly understood. Inspired by the deceased affinity of biotin observed for point mutations of streptavidin at tryptophan residues, we hypothesized that a CH/π interaction may also contribute to the strong interaction between streptavidin and biotin. CH/π interactions were explored and analyzed at the biotin-binding site and at the interface of the subunits by the fragment molecular orbital method (FMO) and extended applications: PIEDA and FMO4. The results show that CH/π interactions are involved in the high affinity for biotin at the binding site of streptavidin. We further suggest that the involvement of CH/π interactions at the subunit interfaces and an extended CH/π network play more critical roles in determining the high affinity, rather than involvement at the binding site. Copyright © 2017 Elsevier Inc. All rights reserved.

Influence of nitrate, sulfate and operational parameters on the bioreduction of perchlorate using an up-flow packed bed reactor at high salinity.

PubMed

Chung, J; Shin, S; Oh, J

2010-05-01

In this study we have investigated whether electron acceptors, such as nitrate or sulphate ions, competitively inhibit the reduction of perchlorate in brine in continuous up-flow packed bed bioreactors. The effect of pH and hydraulic retention time (HRT) on the reduction of perchlorate at high salinity has also been examined. Reduction of perchlorate was found to be only moderately influenced by nitrate (under 163 mg N L-'), implying that there was no significant microbial competition for electron acceptors. As a result of microbial diversity, there were few differences between microbial communities fed with a variety of media, suggesting that most nitrate-reducing bacteria are able to reduce perchlorate at high salinity. Reduction of perchlorate was almost complete at relatively high sulfate levels (1000 mg L(-1)), neutral pH (6-8) and relatively long HRTs (> 10 h).

Understanding the relationship between DOC and nitrate export and dominant rainfall-runoff processes through long-term high frequency measurements

NASA Astrophysics Data System (ADS)

Schwab, Michael; Klaus, Julian; Pfister, Laurent; Weiler, Markus

2016-04-01

Over the past decades, stream sampling protocols for hydro-geochemical parameters were often limited by logistical and technological constraints. While long-term monitoring protocols were typically based on weekly sampling intervals, high frequency sampling was commonly limited to a few single events. In our study, we combined high frequency and long-term measurements to understand the DOC and nitrate behaviour and dynamics for different runoff events and seasons. Our study area is the forested Weierbach catchment (0.47 km2) in Luxembourg. The fractured schist bedrock is covered by cambisol soils. The runoff response of the catchment is characterized by a double peak behaviour. A first discharge peak occurs during or right after a rainfall event (triggered by fast near surface runoff generation processes), while a second delayed peak lasts several days (generated by subsurface flow/ shallow groundwater flow). Peaks in DOC concentrations are closely linked to the first discharge peak, whereas nitrate concentrations follow the second peak. Our observations were carried out with the field deployable instrument spectro::lyser (scan Messtechnik GmbH). This instrument relies on the principles of UV-Vis spectrometry and measures DOC and nitrate concentrations. The measurements were carried out at a high frequency of 15 minutes in situ in the Weierbach creek for more than two years. In addition, a long-term validation was carried out with data obtained from the analysis of water collected with automatic samplers. The long-term, high-frequency measurements allowed us to calculate a complete and detailed balance of DOC and nitrate export over two years. Transport behaviour of the DOC and nitrate showed different dynamics between the first and second hydrograph peaks. DOC is mainly exported during first peaks, while nitrate is mostly exported during the delayed second peaks. In combination with other measurements in the catchment, the long and detailed observations have enabled us to derive relationships between DOC and nitrate export and different catchment states: soil wetness and groundwater levels, precipitation and seasonality. Altogether, the long-term and high-frequency time series provides the opportunity to study DOC and nitrate export without having to just rely only on either a few single event measurements or coarse measurement protocols.

Synthesis and Mass Spectrometric Characterization of Organic Nitrates

NASA Astrophysics Data System (ADS)

Grünert, A.; Woidich, S.; Ballschmiter, K.

2003-04-01

Organic nitrates, as trace constituents in urban air, can be analyzed by adsorptive low volume sampling (LVS) as well as by adsorptive high volume sampling (HVS). Air samples ranging from 25 L to 100 L for the LVS and 100 m3 to 500 m3 for the (HVS) were collected, respectively. Analysis is performed by thermodesorption (LVS) or solvent elution combined with group separation (HVS) using normal-phase HPLC and high resolution capillary gas chromatography with electron capture detection (HRGC-ECD) and mass selective detection (HRGC-MSD). For identification and quantification available reference compounds are required for both methods (1;2). Following numbers of congeners of organic nitrate have been synthesized: 77 monoalkyl nitrates (C1-C16), 43 dialkyl nitrates (C2-C10), 37 hydroxy alkyl nitrates (C2-C8) and 41 carbonyl alkyl nitrates (C3-C12). Alkanes, alkenes, alcohols, ketones and halocarbons have been used as precursors. Characterisation of the reference compounds by retention-data and mass-spectra was performed by high resolution capillary gas chromatography with mass selective detection in the EI- and the NCI (CH4) mode (1-3). EI-ionization leads to the dominating indicator ion NO2+ for organic nitrates with m/z = 46 u. The characteristic fragments with NCI (CH4) show ions at m/z = 46 u and m/z = 62 u, corresponding to NO2- and NO3-. The use of flame ionisation detection (HRGC-FID) and the principle of the molar response for carbon allows the quantitation of reference solutions as the final tool for the determination of the levels and patterns of organic nitrates in urban air samples. (1) J. Kastler: "Analytik, Massenspektrometrie und Vorkommen multifunktioneller Alkylnitrate in belasteter und unbelasteter Atmosphäre" Dr.rer.nat.-Thesis, University of Ulm (1999) (2) G. Werner, J. Kastler, R. Looser, K. Ballschmiter: "Organic Nitrates of Isoprenes as Atmospheric Trace Compounds" Angew. Chem. Int. Ed. (1999) 38(11): 1634-1637 (3) S.Woidich, O. Froscheis, O. Luxenhofer, K. Ballschmiter: "EI- and NCI-mass spectrometry of aryl-alkyl nitrates and their occurrence in urban air" Fresenius J Anal Chem (1999) 364: 91-99

Analysis of the acute response of Galleria mellonella larvae to potassium nitrate.

PubMed

Maguire, Ronan; Kunc, Martin; Hyrsl, Pavel; Kavanagh, Kevin

2017-05-01

Potassium nitrate (E252) is widely used as a food preservative and has applications in the treatment of high blood pressure however high doses are carcinogenic. Larvae of Galleria mellonella were administered potassium nitrate to establish whether the acute effects in larvae correlated with those evident in mammals. Intra-haemocoel injection of potassium nitrate resulted in a significant increase in the density of circulating haemocytes and a small change in the relative proportions of haemocytes but haemocytes showed a reduced fungicidal ability. Potassium nitrate administration resulted in increased superoxide dismutase activity and in the abundance of a range of proteins associated with mitochondrial function (e.g. mitochondrial aldehyde dehydrogenase, putative mitochondrial Mn superoxide dismutase), metabolism (e.g. triosephosphate isomerase, glyceraldehyde 3 phosphate dehydrogenase) and nitrate metabolism (e.g. aliphatic nitrilase, glutathione S-transferase). A strong correlation exists between the toxicity of a range of food preservatives when tested in G. mellonella larvae and rats. In this work a correlation between the effect of potassium nitrate in larvae and mammals is shown and opens the way to the utilization of insects for studying the in vivo acute and chronic toxicity of xenobiotics. Copyright © 2017 Elsevier Inc. All rights reserved.

Enhanced removal of nitrate from water using nZVI@MWCNTs composite: synthesis, kinetics and mechanism of reduction.

PubMed

Babaei, Ali Akbar; Azari, Ali; Kalantary, Roshanak Rezaei; Kakavandi, Babak

2015-01-01

Herein, multi-wall carbon nanotubes (MWCNTs) were used as the carrier of nano-zero valent iron (nZVI) particles to fabricate a composite known as nZVI@MWCNTs. The composite was then characterized and applied in the nitrate removal process in a batch system under anoxic conditions. The influential parameters such as pH, various concentrations of nitrate and composite were investigated within 240 min of the reaction. The mechanism, kinetics and end-products of nitrate reduction were also evaluated. Results revealed that the removal nitrate percentage for nZVI@MWCNTs composite was higher than that of nZVI and MWCNTs alone. Experimental data from nitrate reduction were fitted to the Langmuir-Hinshelwood kinetic model. The values of observed rate constant (kobs) decreased with increasing the initial concentration of nitrate. Our experiments proved that the nitrate removal efficiency was favorable once both high amounts of nZVI@MWCNTs and low concentrations of nitrate were applied. The predominant end-products of the nitrate reduction were ammonium (84%) and nitrogen gas (15%). Our findings also revealed that ZVI@MWCNTs is potentially a good composite for removal/reduction of nitrate from aqueous solutions.

Observations on particulate organic nitrates and unidentified components of NO y

NASA Astrophysics Data System (ADS)

Nielsen, Torben; Egeløv, Axel H.; Granby, Kit; Skov, Henrik

A method to determine the total content of particulate organic nitrates (PON) has been developed and ambient air measurements of PON, NO, N02, HNO3, peroxyacetyl nitrate (PAN), peroxypropionyl nitrate (PPN), gas NOY and particulate inorganic nitrate have been performed in the spring and early summer at an agricultural site in Denmark and compared with measurements of ozone, H 2O 2, SO 2, formic acid, acetic acid and methane sulphonic acid. The gas NO y detector determines the sum NO + NO 2 + HNO 2 + HNO 3 + PAN + PPN + gas phase organic nitrates + 2 × N 2O 5 + NO 3. The content of residual gas NO y ( = gas NO y - NO - NO 2 - HNO 3 - PAN - PPN) was determined and a group of unidentified NO y compounds was found. The phenomenon was observed at a site with relatively high NO x/NO y ratios compared to previous observations in U.S.A. and Canada. The residual gas NO y made up 7 ± 6% of total NOY (total NO y = gas NO y + particulate inorganic nitrate). Residual gas NO y was much higher than the particulate fraction of organic nitrates (PON). PON was only 0.25 ± 0.11% of total NO y. Both residual gas NO y and particulate organic nitrates episodes occurred with elevated concentrations of photochemical oxidants in connection with high-pressure systems suggesting atmospheric processes being the major source. Clean marine air can be discarded as a source for PON and residual gas NO y.

New insights into nitrate dynamics in a karst groundwater system gained from in situ high-frequency optical sensor measurements

USGS Publications Warehouse

Opsahl, Stephen P.; Musgrove, MaryLynn; Slattery, Richard N.

2017-01-01

Understanding nitrate dynamics in groundwater systems as a function of climatic conditions, especially during contrasting patterns of drought and wet cycles, is limited by a lack of temporal and spatial data. Nitrate sensors have the capability for making accurate, high-frequency measurements of nitrate in situ, but have not yet been evaluated for long-term use in groundwater wells. We measured in situ nitrate continuously in two groundwater monitoring wells —one rural and one urban—located in the recharge zone of a productive karst aquifer in central Texas in order to resolve changes that occur over both short-term (hourly to daily) and long-term (monthly to yearly) periods. Nitrate concentrations, measured as nitrate-nitrogen in milligrams per liter (mg/L), during drought conditions showed little or no temporal change as groundwater levels declined. During aquifer recharge, extremely rapid changes in concentration occurred at both wells as documented by hourly data. At both sites, nitrate concentrations were affected by recharging surface water as evidenced by nitrate concentrations in groundwater recharge (0.8–1.3 mg/L) that were similar to previously reported values for regional recharging streams. Groundwater nitrate concentrations responded differently at urban and rural sites during groundwater recharge. Concentrations at the rural well (approximately 1.0 mg/L) increased as a result of higher nitrate concentrations in groundwater recharge relative to ambient nitrate concentrations in groundwater, whereas concentrations at the urban well (approximately 2.7 mg/L) decreased as a result of the dilution of higher ambient nitrate concentrations relative to those in groundwater recharge. Notably, nitrate concentrations decreased to as low as 0.8 mg/L at the urban site during recharge but postrecharge concentrations exceeded 3.0 mg/L. A return to higher nitrate concentrations postrecharge indicates mobilization of a localized source of elevated nitrate within the urbanized area of the aquifer. Changes in specific conductance were observed at both sites during groundwater recharge, and a significant correlation between specific conductance and nitrate (correlation coefficient [R] = 0.455) was evident at the urban site where large (3-fold) changes in nitrate occurred. Nitrate concentrations and specific conductance measured during a depth profile indicated that the water column was generally homogeneous as expected for this karst environment, but changes were observed in the most productive zone of the aquifer that might indicate some heterogeneity within the complex network of flow paths. Resolving the timing and magnitude of changes and characterizing fine-scale vertical differences would not be possible using conventional sampling techniques. The patterns observed in situ provided new insight into the dynamic nature of nitrate in a karst groundwater system.

Factors influencing protein tyrosine nitration – structure-based predictive models

PubMed Central

Bayden, Alexander S.; Yakovlev, Vasily A.; Graves, Paul R.; Mikkelsen, Ross B.; Kellogg, Glen E.

2010-01-01

Models for exploring tyrosine nitration in proteins have been created based on 3D structural features of 20 proteins for which high resolution X-ray crystallographic or NMR data are available and for which nitration of 35 total tyrosines has been experimentally proven under oxidative stress. Factors suggested in previous work to enhance nitration were examined with quantitative structural descriptors. The role of neighboring acidic and basic residues is complex: for the majority of tyrosines that are nitrated the distance to the heteroatom of the closest charged sidechain corresponds to the distance needed for suspected nitrating species to form hydrogen bond bridges between the tyrosine and that charged amino acid. This suggests that such bridges play a very important role in tyrosine nitration. Nitration is generally hindered for tyrosines that are buried and for those tyrosines where there is insufficient space for the nitro group. For in vitro nitration, closed environments with nearby heteroatoms or unsaturated centers that can stabilize radicals are somewhat favored. Four quantitative structure-based models, depending on the conditions of nitration, have been developed for predicting site-specific tyrosine nitration. The best model, relevant for both in vitro and in vivo cases predicts 30 of 35 tyrosine nitrations (positive predictive value) and has a sensitivity of 60/71 (11 false positives). PMID:21172423

[Role of hemoglobin affinity to oxygen in adaptation to hypoxemia].

PubMed

Kwasiborski, Przemysław Jerzy; Kowalczyk, Paweł; Zieliński, Jakub; Przybylski, Jacek; Cwetsch, Andrzej

2010-04-01

One of the basic mechanisms of adapting to hypoxemia is a decrease in the affinity of hemoglobin for oxygen. This process occurs mainly due to the increased synthesis of 2,3-diphosphoglycerate (2,3-DPG) in the erythrocytes, as well as through the Bohr effect. Hemoglobin with decreased affinity for oxygen increases the oxygenation of tissues, because it gives up oxygen more easily during microcirculation. In foetal circulation, however, at a partial oxygen pressure (pO2) of 25 mmHg in the umbilical vein, the oxygen carrier is type F hemoglobin which has a high oxygen affinity. The commonly accepted role for hemoglobin F is limited to facilitating diffusion through the placenta. Is fetal life the only moment when haemoglobin F is useful? THE AIM OF STUDY was to create a mathematical model, which would answer the question at what conditions an increase, rather than a decrease, in haemoglobin oxygen affinity is of benefit to the body. Using the kinetics of dissociation of oxygen from hemoglobin described by the Hill equation as the basis for further discussion, we created a mathematical model describing the pO2 value in the microcirculatory system and its dependence on arterial blood pO2. The calculations were performed for hemoglobin with low oxygen affinity (adult type) and high-affinity hemoglobin (fetal type). The modelling took into account both physiological and pathological ranges of acid-base equilibrium and tissue oxygen extraction parameters. It was shown that for the physiological range of acid-base equilibrium and the resting level of tissue oxygen extraction parameters, with an arterial blood pO2 of 26.8 mmHg, the higher-affinity hemoglobin becomes the more effective oxygen carrier. It was also demonstrated that the arterial blood pO2, below which the high-affinity hemoglobin becomes the more effective carrier, is dependent on blood pH and the difference between the arterial and venous oxygen saturation levels. Simulations performed for the pathological states showed that acidosis and increased tissue oxygen demand lead to a broadened arterial blood pO2 range, in which the high-affinity hemoglobin is more efficient. Contrary to the widely held view that the only response to hypoxemia is a decrease in haemoglobin oxygen affinity, it was shown that under extreme hypoxemic conditions, an increased haemoglobin oxygen affinity improves the oxygenation of tissues. It was also shown that the dominance of hemoglobin with a high oxygen affinity rapidly exceeds hemoglobin with low oxygen affinity in the case of acidosis with its accompanying high tissue oxygen extraction. In cases of extreme disruptions of the acid-base equilibrium, the dominance of high-oxygen-affinity hemoglobin spans over the entire possible range of pO2 in arterial blood.

Affinity proteomics to study endogenous protein complexes: Pointers, pitfalls, preferences and perspectives

PubMed Central

LaCava, John; Molloy, Kelly R.; Taylor, Martin S.; Domanski, Michal; Chait, Brian T.; Rout, Michael P.

2015-01-01

Dissecting and studying cellular systems requires the ability to specifically isolate distinct proteins along with the co-assembled constituents of their associated complexes. Affinity capture techniques leverage high affinity, high specificity reagents to target and capture proteins of interest along with specifically associated proteins from cell extracts. Affinity capture coupled to mass spectrometry (MS)-based proteomic analyses has enabled the isolation and characterization of a wide range of endogenous protein complexes. Here, we outline effective procedures for the affinity capture of protein complexes, highlighting best practices and common pitfalls. PMID:25757543

Legacy Nitrate Impacts on Groundwater and Streams

NASA Astrophysics Data System (ADS)

Tesoriero, A. J.; Juckem, P. F.; Miller, M. P.

2017-12-01

Decades of recharge of high-nitrate groundwater have created a legacy—a mass of high-nitrate groundwater—that has implications for future nitrate concentrations in groundwater and in streams. In the United States, inorganic nitrogen fertilizer applications to the land surface have increased ten-fold since 1950, resulting in sharp increases in nitrate concentrations in recharging groundwater, which pose a risk to deeper groundwater and streams. This study assesses the factors that control time lags and eventual concentrations of legacy nitrate in groundwater and streams. Results from the USGS National Water-Quality Assessment Project are presented which elucidate nitrate trends in recharging groundwater, delineate redox zones and assess groundwater and stream vulnerability to legacy nitrate sources on a regional scale. This study evaluated trends and transformations of agricultural chemicals based on groundwater age and water chemistry data along flow paths from recharge areas to streams at 20 study sites across the United States. Median nitrate recharge concentrations in these agricultural areas have increased markedly over the last 50 years, from 4 to 7.5 mg N/L. The effect that nitrate accumulation in shallow aquifers will have on drinking water quality and stream ecosystems is dependent on the redox zones encountered along flow paths and on the age distribution of nitrate discharging to supply wells and streams. Delineating redox zones on a regional scale is complicated by the spatial variability of reaction rates. To overcome this limitation, we applied logistic regression and machine learning techniques to predict the probability of a specific redox condition in groundwater in the Chesapeake Bay watershed and the Fox-Wolf-Peshtigo study area in Wisconsin. By relating redox-active constituent concentrations in groundwater samples to indicators of residence time and/or electron donor availability, we were able to delineate redox zones on a regional scale - an important indicator of groundwater vulnerability and the vulnerability of streams to legacy nitrate sources.

No Effect of Acute and 6-Day Nitrate Supplementation on VO2 and Time-Trial Performance in Highly Trained Cyclists.

PubMed

Nyakayiru, Jean M; Jonvik, Kristin L; Pinckaers, Philippe J M; Senden, Joan; van Loon, Luc J C; Verdijk, Lex B

2017-02-01

While the majority of studies reporting ergogenic effects of dietary nitrate have used a multiday supplementation protocol, some studies suggest that a single dose of dietary nitrate before exercise can also improve subsequent performance. We aimed to compare the impact of acute and 6-day sodium nitrate supplementation on oxygen uptake (V̇O 2 ) and time-trial performance in trained cyclists. Using a randomized, double-blind, cross-over design, 17 male cyclists (25 ± 4 y, V̇O 2peak 65 ± 4 ml·kg -1 ·min -1 , W max 411 ± 35 W) were subjected to 3 different trials; 5 days placebo and 1 day sodium nitrate supplementation (1-DAY); 6 days sodium nitrate supplementation (6-DAY); 6 days placebo supplementation (PLA). Nitrate was administered as 1097 mg sodium nitrate providing 800 mg (~12.9 mmol) nitrate per day. Three hours after ingestion of the last supplemental bolus, indirect calorimetry was performed while subjects performed 30 min of exercise at 45% W max and 30 min at 65% W max on a cycle ergometer, followed by a 10 km time-trial. Immediately before exercise, plasma [nitrate] and [nitrite] increased to a similar extent during the 6-DAY and 1-DAY trial, but not with PLA (plasma nitrite: 501 ± 205, 553 ± 278, and 239 ± 74 nM, respectively; p < .001). No differences were observed between interventions in V̇O 2 during submaximal exercise, or in time to complete the time-trial (6-DAY: 1004 ± 61, 1-DAY: 1022 ± 72, PLA: 1017 ± 71 s; p = .28). We conclude that both acute and 6-days of sodium nitrate supplementation do not alter V̇O 2 during submaximal exercise or improve time-trial performance in highly trained cyclists, despite increasing plasma [nitrate] and [nitrite].

Stable isotope evidence for an atmospheric origin of desert nitrate deposits in northern Chile and southern California, U.S.A.

USGS Publications Warehouse

Böhlke, J.K.; Ericksen, G.E.; Revesz, K.

1997-01-01

Natural surficial accumulations of nitrate-rich salts in the Atacama Desert, northern Chile, and in the Death Valley region of the Mojave Desert, southern California, are well known, but despite many geologic and geochemical studies, the origins of the nitrates have remained controversial. N and O isotopes in nitrate, and S isotopes in coexisting soluble sulfate, were measured to determine if some proposed N sources could be supported or rejected, and to determine if the isotopic signature of these natural deposits could be used to distinguish them from various types of anthropogenic nitrate contamination that might be found in desert groundwaters. High-grade caliche-type nitrate deposits from both localities have ??15N values that range from -5 to +5???, but are mostly near 0???. Values of ??15N near 0??? are consistent with either bulk atmospheric N deposition or microbial N fixation as major sources of the N in the deposits. ??18O values of those desert nitrates with ??15N near 0??? range from about +31 to + 50??? (V-SMOW), significantly higher than that of atmospheric O2 (+ 23.5???). Such high values of ??18O are considered unlikely to result entirely from nitrification of reduced N, but rather resemble those of modern atmospheric nitrate in precipitation from some other localities. Assuming that limited modern atmospheric isotope data are applicable to the deposits, and allowing for nitrification of co-deposited ammonium, it is estimated that the fraction of the nitrate in the deposits that could be accounted for isotopically by atmospheric N deposition may be at least 20% and possibly as much as 100%. ??34S values are less diagnostic but could also be consistent with atmospheric components in some of the soluble sulfates associated with the deposits. The stable isotope data support the hypothesis that some high-grade caliche-type nitrate-rich salt deposits in some of the Earth's hyperarid deserts represent long-term accumulations of atmospheric deposition (possibly in the order of 104 yr for the Death Valley region. 107 yr for the Atacama Desert) in the relative absence of soil leaching or biologic cycling. The combined N and O isotope signature of the nitrate in these deposits is significantly different from those of many other natural and anthropogenic sources of nitrate.

HIGH-AFFINITY T CELL RECEPTOR DIFFERENTIATES COGNATE PEPTIDE-MHC AND ALTERED PEPTIDE LIGANDS WITH DISTINCT KINETICS AND THERMODYNAMICS

PubMed Central

Persaud, Stephen P.; Donermeyer, David L.; Weber, K. Scott; Kranz, David M.; Allen, Paul M.

2010-01-01

Interactions between the T cell receptor and cognate peptide-MHC are crucial initiating events in the adaptive immune response. These binding events are highly specific yet occur with micromolar affinity. Even weaker interactions between TCR and self-pMHC complexes play critical regulatory roles in T cell development, maintenance and coagonist activity. Due to their low affinity, the kinetics and thermodynamics of such weak interactions are difficult to study. In this work, we used M15, a high-affinity TCR engineered from the 3.L2 TCR system, to study the binding properties, thermodynamics, and specificity of two altered peptide ligands (APLs). Our affinity measurements of the high-affinity TCR support the view that the wild type TCR binds these APLs in the millimolar affinity range, and hence very low affinities can still elicit biological functions. Finally, single methylene differences among the APLs gave rise to strikingly different binding thermodynamics. These minor changes in the pMHC antigen were associated with significant and unpredictable changes in both the entropy and enthalpy of the reaction. As the identical TCR was analyzed with several structurally similar ligands, the distinct thermodynamic binding profiles provide a mechanistic perspective on how exquisite antigen specificity is achieved by the T cell receptor. PMID:20334923

Quantifying in-stream nitrate reaction rates using continuously-collected water quality data

Treesearch

Matthew Miller; Anthony Tesoriero; Paul Capel

2016-01-01

High frequency in situ nitrate data from three streams of varying hydrologic condition, land use, and watershed size were used to quantify the mass loading of nitrate to streams from two sources – groundwater discharge and event flow – at a daily time step for one year. These estimated loadings were used to quantify temporally-variable in-stream nitrate processing ...

Mechanisms underlying export of N from high-elevation catchments during seasonal transitions

USGS Publications Warehouse

Sickman, J.O.; Leydecker, A.L.; Chang, Cecily C.Y.; Kendall, C.; Melack, J.M.; Lucero, D.M.; Schimel, J.

2003-01-01

Mechanisms underlying catchment export of nitrogen (N) during seasonal transitions (i.e., winter to spring and summer to autumn) were investigated in high-elevation catchments of the Sierra Nevada using stable isotopes of nitrate and water, intensive monitoring of stream chemistry and detailed catchment N-budgets. We had four objectives: (1) determine the relative contribution of snowpack and soil nitrate to the spring nitrate pulse, (2) look for evidence of biotic control of N losses at the catchment scale, (3) examine dissolved organic nitrogen (DON) export patterns to gain a better understanding of the biological and hydrological controls on DON loss, and (4) examine the relationship between soil physico-chemical conditions and N export. At the Emerald Lake watershed, nitrogen budgets and isotopic analyses of the spring nitrate pulse indicate that 50 to 70% of the total nitrate exported during snowmelt (ca. April to July) is derived from catchment soils and talus; the remainder is snowpack nitrate. The spring nitrate pulse occurred several weeks after the start of snowmelt and was different from export patterns of less biologically labile compounds such as silica and DON suggesting that: (1) nitrate is produced and released from soils only after intense flushing has occurred and (2) a microbial N-sink is operating in catchment soils during the early stages of snowmelt. DON concentrations varied less than 20-30% during snowmelt, indicating that soil processes tightly controlled DON losses.

Microbially Enhanced Oil Recovery by Sequential Injection of Light Hydrocarbon and Nitrate in Low- And High-Pressure Bioreactors.

PubMed

Gassara, Fatma; Suri, Navreet; Stanislav, Paul; Voordouw, Gerrit

2015-10-20

Microbially enhanced oil recovery (MEOR) often involves injection of aqueous molasses and nitrate to stimulate resident or introduced bacteria. Use of light oil components like toluene, as electron donor for nitrate-reducing bacteria (NRB), offers advantages but at 1-2 mM toluene is limiting in many heavy oils. Because addition of toluene to the oil increased reduction of nitrate by NRB, we propose an MEOR technology, in which water amended with light hydrocarbon below the solubility limit (5.6 mM for toluene) is injected to improve the nitrate reduction capacity of the oil along the water flow path, followed by injection of nitrate, other nutrients (e.g., phosphate) and a consortium of NRB, if necessary. Hydrocarbon- and nitrate-mediated MEOR was tested in low- and high-pressure, water-wet sandpack bioreactors with 0.5 pore volumes of residual oil in place (ROIP). Compared to control bioreactors, those with 11-12 mM of toluene in the oil (gained by direct addition or by aqueous injection) and 80 mM of nitrate in the aqueous phase produced 16.5 ± 4.4% of additional ROIP (N = 10). Because toluene is a cheap commodity chemical, HN-MEOR has the potential to be a cost-effective method for additional oil production even in the current low oil price environment.

Let's get specific: the relationship between specificity and affinity.

PubMed

Eaton, B E; Gold, L; Zichi, D A

1995-10-01

The factors that lead to high-affinity binding are a good fit between the surfaces of the two molecules in their ground state and charge complementarity. Exactly the same factors give high specificity for a target. We argue that selection for high-affinity binding automatically leads to highly specific binding. This principle can be used to simplify screening approaches aimed at generating useful drugs.

High-Resolution in Situ Measurement of Nitrate in Runoff from the Greenland Ice Sheet.

PubMed

Beaton, Alexander D; Wadham, Jemma L; Hawkings, Jon; Bagshaw, Elizabeth A; Lamarche-Gagnon, Guillaume; Mowlem, Matthew C; Tranter, Martyn

2017-11-07

We report the first in situ high-resolution nitrate time series from two proglacial meltwater rivers draining the Greenland Ice Sheet, using a recently developed submersible analyzer based on lab-on-chip (LOC) technology. The low sample volume (320 μL) required by the LOC analyzer meant that low concentration (few micromolar to submicromolar), highly turbid subglacial meltwater could be filtered and colorimetrically analyzed in situ. Nitrate concentrations in rivers draining Leverett Glacier in southwest Greenland and Kiattuut Sermiat in southern Greenland exhibited a clear diurnal signal and a gradual decline at the commencement of the melt season, displaying trends that would not be discernible using traditional daily manual sampling. Nitrate concentrations varied by 4.4 μM (±0.2 μM) over a 10 day period at Kiattuut Sermiat and 3.0 μM (±0.2 μM) over a 14 day period at Leverett Glacier. Marked changes in nitrate concentrations were observed when discharge began to increase. High-resolution in situ measurements such as these have the potential to significantly advance the understanding of nutrient cycling in remote systems, where the dynamics of nutrient release are complex but are important for downstream biogeochemical cycles.

Effects of melanin on the accumulation of exopolysaccharides by Aureobasidium pullulans grown on nitrate.

PubMed

Zheng, Weifa; Campbell, Bradley S; McDougall, Barbara M; Seviour, Robert J

2008-11-01

Aureobasidium pullulans produced pullulan and melanin when grown in medium containing low nitrate levels. With high nitrate concentrations, however, this fungus produced a mixture of exopolysaccharides (EPS) without melanin synthesis. At 0.78 g l(-1) N as nitrate, where no melanin synthesis occurred, maximum EPS yields reached 6.92 g l(-1) and then decreased to the final yield of 2.36 g l(-1). Following melanin addition (0.1 g l(-1)), yields reached 7.02 g l(-1) at 48 h and fell to a final yield of 5.21 g l(-1). The EPS produced in high nitrate medium contained both pullulan and (1-->3)-beta-glucan, but only pullulan was produced with melanin-supplementation. With melanin addition a doubling of (1-->3)-beta-glucanase activity was observed in high nitrate medium compared to that without supplementation. On the other hand amylolytic activities disappeared in medium with melanin production or addition. Culture filtrates sustained a higher reducing capacity (RC) when melanin was present. Low RC appeared to reduce (1-->3)-beta-glucanase activity and increase amylolytic activities. Thus, higher RC appears to inhibit production/activity of amylose-degrading enzymes capable of degrading pullulan, and stimulates (1-->3)-beta-glucanase synthesis/activity, leading to a preferential accumulation of pullulan.

Chemometric evaluation of nitrate contamination in the groundwater of a hard rock area in Dharapuram, south India

NASA Astrophysics Data System (ADS)

Sajil Kumar, P. J.; Jegathambal, P.; James, E. J.

2014-12-01

This paper presents the results of investigations on groundwater nitrate contamination in the Dharapuram area of Tamil Nadu in south India as a primary step to initiate denitrification. Groundwater samples were collected from 26 selected locations during the pre-monsoon season in July 2010 and analysed for nitrate and other water quality parameters. Two important water types were identified, viz. Ca-Na-HCO3 and mixed Ca-Mg-Cl. It is found that the majority of samples possess high nitrate concentration; 57 % of samples exceeded the permissible limit of Indian (45 mg/L) and WHO (50 mg/L) drinking water standard. Spatial distribution map of NO3 suggested that major contamination was observed in the SW and NW parts of the study area. This result was in agreement with the corresponding land-use pattern in this study area. Denitrification process at greater depths was evident from the negative correlation between NO3 and well depth. The sources and controlling factors of high nitrate were investigated using cross plots of NO3 with other selected hydrochemical parameters. Positive correlation for NO3 was observed with EC, K, Cl and SO4. This analysis was capable of differentiating the various sources of nitrate in groundwater. The major sources of nitrate contamination are identified as areas of high fertilizer application, sewages and animal waste dumping yards. Regulation of these pollutant sources with appropriate and cost-effective denitrification process can restore the water quality in this area.

Pig slurry application and irrigation effects on nitrate leaching in Mediterranean soil lysimeters.

PubMed

Daudén, A; Quílez, D; Vera, M V

2004-01-01

Land application of animal manures, such as pig slurry (PS), is a common practice in intensive-farming agriculture. However, this practice has a pitfall consisting of the loss of nutrients, in particular nitrate, toward water courses. The objective of this study was to evaluate nitrate leaching for three application rates of pig slurry (50, 100, and 200 Mg ha(-1)) and a control treatment of mineral fertilizer (275 kg N ha(-1)) applied to corn grown in 10 drainage lysimeters. The effects of two irrigation regimes (low vs. high irrigation efficiency) were also analyzed. In the first two irrigation events, drainage NO(3)-N concentrations as high as 145 and 69 mg L(-1) were measured in the high and moderate PS rate treatments, respectively, in the low irrigation efficiency treatments. This indicates the fast transformation of the PS ammonium into nitrate and the subsequent leaching of the transformed nitrate. Drainage NO(3)-N concentration and load increased linearly by 0.69 mg NO(3)-N L(-1) and 4.6 kg NO(3)-N ha(-1), respectively, for each 10 kg N ha(-1) applied over the minimum of 275 kg N ha(-1). An increase in irrigation efficiency did not induce a significant increase of leachate concentration and the amount of nitrate leached decreased about 65%. Application of low PS doses before sowing complemented with sidedressing N application and a good irrigation management are the key factors to reduce nitrate contamination of water courses.

Three-component access to pyrroles promoted by the CAN-silver nitrate system under high-speed vibration milling conditions: a generalization of the Hantzsch pyrrole synthesis.

PubMed

Estévez, Verónica; Villacampa, Mercedes; Menéndez, J Carlos

2013-01-21

A sequential multicomponent process involving the high-speed vibration milling of ketones with N-iodosuccinimide and p-toluenesulfonic acid, followed by addition of a mixture of primary amines, β-dicarbonyl compounds, cerium(IV) ammonium nitrate and silver nitrate afforded polysubstituted, functionalized pyrroles. This one-pot, solid-state process can be considered as the coupling of an α-iodoketone preparation with a general version of the classical Hantzsch pyrrole synthesis.

Thyroid Cancer Induction: Nitrates as Independent Risk Factors or Risk Modulators after Radiation Exposure, with a Focus on the Chernobyl Accident

PubMed Central

Drozd, Valentina M.; Branovan, Igor; Shiglik, Nikolay; Biko, Johannes; Reiners, Christoph

2018-01-01

In recent decades, differentiated thyroid cancer (DTC) incidence has been increasing worldwide. The important contributions to this phenomenon of “overdiagnosis” driven by wider use of improved ultrasound systems are amply documented, notwithstanding the “real” carcinogenic effects of ionizing radiation, e.g., from the Chernobyl accident or health care interventions. Less well understood is the role of nitrates – as environmental pollutants, in diet, and in medication – in thyroid carcinogenesis. Increasing exposure to nitrates is associated with rising incidence of esophageal, stomach, bladder, and colon cancers. Recent data suggest that in agricultural areas with higher mean nitrate levels in groundwater, DTC risk is also elevated. Our work in Belarus after Chernobyl has shown that children in districts with high nitrate concentrations in drinking water had significantly higher thyroid cancer incidence after irradiation than did their counterparts in areas with lower nitrate concentrations. Notwithstanding thyroid shielding, increasing use of computed tomography and dental X-rays heightens radiation exposure of the salivary glands in the general population, especially in children and adolescents. When nitrate intake is increased, salivary gland irradiation may potentially result in carcinogenic elevations in plasma nitric oxide concentrations. In conclusion, excess nitrate intake seems to be an independent risk factor for DTC. Additionally, we hypothesize from our data that high nitrate levels modulate the carcinogenic effect of radiation on the thyroid. Cohort studies, case-control studies, or both, are needed to quantify the effects of nitrates on DTC risk in the presence or absence of radiation exposure, e.g., that associated with diagnostic or therapeutic health care interventions. PMID:29594057

Probability-based nitrate contamination map of groundwater in Kinmen.

PubMed

Liu, Chen-Wuing; Wang, Yeuh-Bin; Jang, Cheng-Shin

2013-12-01

Groundwater supplies over 50% of drinking water in Kinmen. Approximately 16.8% of groundwater samples in Kinmen exceed the drinking water quality standard (DWQS) of NO3 (-)-N (10 mg/L). The residents drinking high nitrate-polluted groundwater pose a potential risk to health. To formulate effective water quality management plan and assure a safe drinking water in Kinmen, the detailed spatial distribution of nitrate-N in groundwater is a prerequisite. The aim of this study is to develop an efficient scheme for evaluating spatial distribution of nitrate-N in residential well water using logistic regression (LR) model. A probability-based nitrate-N contamination map in Kinmen is constructed. The LR model predicted the binary occurrence probability of groundwater nitrate-N concentrations exceeding DWQS by simple measurement variables as independent variables, including sampling season, soil type, water table depth, pH, EC, DO, and Eh. The analyzed results reveal that three statistically significant explanatory variables, soil type, pH, and EC, are selected for the forward stepwise LR analysis. The total ratio of correct classification reaches 92.7%. The highest probability of nitrate-N contamination map presents in the central zone, indicating that groundwater in the central zone should not be used for drinking purposes. Furthermore, a handy EC-pH-probability curve of nitrate-N exceeding the threshold of DWQS was developed. This curve can be used for preliminary screening of nitrate-N contamination in Kinmen groundwater. This study recommended that the local agency should implement the best management practice strategies to control nonpoint nitrogen sources and carry out a systematic monitoring of groundwater quality in residential wells of the high nitrate-N contamination zones.

Unmasking of CD22 Co-receptor on Germinal Center B-cells Occurs by Alternative Mechanisms in Mouse and Man*

PubMed Central

Macauley, Matthew S.; Kawasaki, Norihito; Peng, Wenjie; Wang, Shui-Hua; He, Yuan; Arlian, Britni M.; McBride, Ryan; Kannagi, Reiji; Khoo, Kay-Hooi; Paulson, James C.

2015-01-01

CD22 is an inhibitory B-cell co-receptor whose function is modulated by sialic acid (Sia)-bearing glycan ligands. Glycan remodeling in the germinal center (GC) alters CD22 ligands, with as yet no ascribed biological consequence. Here, we show in both mice and humans that loss of high affinity ligands on GC B-cells unmasks the binding site of CD22 relative to naive and memory B-cells, promoting recognition of trans ligands. The conserved modulation of CD22 ligands on GC B-cells is striking because high affinity glycan ligands of CD22 are species-specific. In both species, the high affinity ligand is based on the sequence Siaα2–6Galβ1–4GlcNAc, which terminates N-glycans. The human ligand has N-acetylneuraminic acid (Neu5Ac) as the sialic acid, and the high affinity ligand on naive B-cells contains 6-O-sulfate on the GlcNAc. On human GC B-cells, this sulfate modification is lost, giving rise to lower affinity CD22 ligands. Ligands of CD22 on naive murine B-cells do not contain the 6-O-sulfate modification. Instead, the high affinity ligand for mouse CD22 has N-glycolylneuraminic acid (Neu5Gc) as the sialic acid, which is replaced on GC B-cells with Neu5Ac. Human naive and memory B-cells express sulfated glycans as high affinity CD22 ligands, which are lost on GC B-cells. In mice, Neu5Gc-containing glycans serve as high affinity CD22 ligands that are replaced by Neu5Ac-containing glycans on GC B-cells. Our results demonstrate that loss of high affinity CD22 ligands on GC B-cells occurs in both mice and humans through alternative mechanisms, unmasking CD22 relative to naive and memory B-cells. PMID:26507663

Unmasking of CD22 Co-receptor on Germinal Center B-cells Occurs by Alternative Mechanisms in Mouse and Man.

PubMed

Macauley, Matthew S; Kawasaki, Norihito; Peng, Wenjie; Wang, Shui-Hua; He, Yuan; Arlian, Britni M; McBride, Ryan; Kannagi, Reiji; Khoo, Kay-Hooi; Paulson, James C

2015-12-11

CD22 is an inhibitory B-cell co-receptor whose function is modulated by sialic acid (Sia)-bearing glycan ligands. Glycan remodeling in the germinal center (GC) alters CD22 ligands, with as yet no ascribed biological consequence. Here, we show in both mice and humans that loss of high affinity ligands on GC B-cells unmasks the binding site of CD22 relative to naive and memory B-cells, promoting recognition of trans ligands. The conserved modulation of CD22 ligands on GC B-cells is striking because high affinity glycan ligands of CD22 are species-specific. In both species, the high affinity ligand is based on the sequence Siaα2-6Galβ1-4GlcNAc, which terminates N-glycans. The human ligand has N-acetylneuraminic acid (Neu5Ac) as the sialic acid, and the high affinity ligand on naive B-cells contains 6-O-sulfate on the GlcNAc. On human GC B-cells, this sulfate modification is lost, giving rise to lower affinity CD22 ligands. Ligands of CD22 on naive murine B-cells do not contain the 6-O-sulfate modification. Instead, the high affinity ligand for mouse CD22 has N-glycolylneuraminic acid (Neu5Gc) as the sialic acid, which is replaced on GC B-cells with Neu5Ac. Human naive and memory B-cells express sulfated glycans as high affinity CD22 ligands, which are lost on GC B-cells. In mice, Neu5Gc-containing glycans serve as high affinity CD22 ligands that are replaced by Neu5Ac-containing glycans on GC B-cells. Our results demonstrate that loss of high affinity CD22 ligands on GC B-cells occurs in both mice and humans through alternative mechanisms, unmasking CD22 relative to naive and memory B-cells. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Differential effects of ethanol and other inducers of drug metabolism on the two forms of hamster liver microsomal aniline hydroxylase.

PubMed

McCoy, G D

1980-03-01

The aniline hydroxylase activity of microsomes isolated from hamster liver can be differentiated kinetically into high affinity (low K(m), form I) and low affinity (high K(m), form II) forms. Microsomes isolated from uninduced animals contain slightly more form I activity. The activity of the low affinity form (form II) is preferentially enhanced by Aroclor or 3-methylcholanthrene treatment, while phenobarbital treatment increases the activity of both forms. Chronic ethanol consumption results in enhancement of only the high affinity form (form I).

Evolutionary processes and sources of high-nitrate haze episodes over Beijing, Spring.

PubMed

Yang, Ting; Sun, Yele; Zhang, Wei; Wang, Zifa; Liu, Xingang; Fu, Pingqing; Wang, Xiquan

2017-04-01

Rare and consecutive high-nitrate haze pollution episodes were observed in Beijing in spring 2012. We present detailed characterization of the sources and evolutionary mechanisms of this haze pollution, and focus on an episode that occurred between 15 and 26 April. Submicron aerosol species were found to be substantially elevated during haze episodes, and nitrates showed the largest increase and occupation (average: 32.2%) in non-refractory submicron particles (NR-PM 1 ), which did not occur in other seasons as previously reported. The haze episode (HE) was divided into three sub-episodes, HEa, HEb, and HEc. During HEa and HEc, a shallow boundary layer, stagnant meteorological conditions, and high humidity favored the formation of high-nitrate concentrations, which were mainly produced by three different processes - daytime photochemical production, gas-particle partitioning, and nighttime heterogeneous reactions - and the decline in visibility was mainly induced by NR-PM 1. However, unlike HEa and HEc, during HEb, the contribution of high nitrates was partly from the transport of haze from the southeast of Beijing - the transport pathway was observed at ~800-1000m by aerosol Lidar - and the decline in visibility during HEb was primarily caused by PM 2.5 . Our results provide useful information for air quality improvement strategies in Beijing during Spring. Copyright © 2016. Published by Elsevier B.V.

Removal of highly elevated nitrate from drinking water by pH-heterogenized heterotrophic denitrification facilitated with ferrous sulfide-based autotrophic denitrification.

PubMed

Huang, Bin; Chi, Guangyu; Chen, Xin; Shi, Yi

2011-11-01

The performance of acetic acid-supported pH-heterogenized heterotrophic denitrification (HD) facilitated with ferrous sulfide-based autotrophic denitrification (AD) was investigated in upflow activated carbon-packed column reactors for reliable removal of highly elevated nitrate (42 mg NO(3)-Nl(-1)) in drinking water. The use of acetic acid as substrate provided sufficient internal carbon dioxide to completely eliminate the need of external pH adjustment for HD, but simultaneously created vertically heterogenized pH varying from 4.8 to 7.8 in the HD reactor. After 5-week acclimation, the HD reactor developed a moderate nitrate removal capacity with about one third of nitrate removal occurring in the acidic zone (pH 4.8-6.2). To increase the treatment reliability, acetic acid-supported HD was operated under 10% carbon limitation to remove >85% of nitrate, and ferrous sulfide-based AD was supplementally operated to remove residual nitrate and formed nitrite without excess of soluble organic carbon, nitrite or sulfate in the final effluent. Copyright © 2011 Elsevier Ltd. All rights reserved.

[Erythropoietin influence on oxygen transport function of blood and prooxidant/antioxidant balance in rabbits under lipopolysaccharide injection].

PubMed

Zinchuk, V V; Shul'ga, E V; Guliaĭ, I E

2010-01-01

We aimed to study the erythropoietin influence on oxygen transport function of blood and prooxidant/antioxidant balance in rabbits under lipopolysaccharide injection. Recombinant human erythropoietin-alpha was administered intraperitoneally in the dose 1000 U/kg 30 minutes before intravenous injection of 500 mkg/kg lipopolysaccharide from E. coli. After 12 hours, blood samples were collected for the assessment of oxygen transport function of blood; nitrate/nitrite levels and tissue samples were collected for measurement of conjugated dienes, malondialdehyde, alpha-tocopherol and catalase. Erythropoietin improves parameters of oxygen transport function of blood, increases hemoglobin-oxygen affinity through the NO-dependent mechanism, reduces activity of free radical processes, and increases antioxidant protection under lipopolysaccharide injection.

Assessment of the Density Functional Tight Binding Method for Protic Ionic Liquids

PubMed Central

2015-01-01

Density functional tight binding (DFTB), which is ∼100–1000 times faster than full density functional theory (DFT), has been used to simulate the structure and properties of protic ionic liquid (IL) ions, clusters of ions and the bulk liquid. Proton affinities for a wide range of IL cations and anions determined using DFTB generally reproduce G3B3 values to within 5–10 kcal/mol. The structures and thermodynamic stabilities of n-alkyl ammonium nitrate clusters (up to 450 quantum chemical atoms) predicted with DFTB are in excellent agreement with those determined using DFT. The IL bulk structure simulated using DFTB with periodic boundary conditions is in excellent agreement with published neutron diffraction data. PMID:25328497

Cu2+ ions as a paramagnetic probe to study the surface chemical modification process of layered double hydroxides and hydroxide salts with nitrate and carboxylate anions.

PubMed

Arizaga, Gregorio Guadalupe Carbajal; Mangrich, Antonio Salvio; Wypych, Fernando

2008-04-01

A layered zinc hydroxide nitrate (Zn5(OH)8(NO3)2.2H2O) and a layered double hydroxide (Zn/Al-NO3) were synthesized by coprecipitation and doped with different amounts of Cu2+ (0.2, 1, and 10 mol%), as paramagnetic probe. Although the literature reports that the nitrate ion is free (with D3h symmetry) between the layers of these two structures, the FTIR spectra of two zinc hydroxide nitrate samples show the C2v symmetry for the nitrate ion, whereas the g ||/A || value in the EPR spectra of Cu2+ is high. This fact suggests bonding of some nitrate ions to the layers of the zinc hydroxide nitrate. The zinc hydroxide nitrate was used as matrix in the intercalation reaction with benzoate, o-chlorobenzoate, and o-iodobenzoate ions. FTIR spectra confirm the ionic exchange reaction and the EPR spectroscopy reveals bonding of the organic ions to the inorganic layers of the zinc hydroxide nitrate, while the layered double hydroxides show only exchange reactions.

Systems analysis of transcriptome data provides new hypotheses about Arabidopsis root response to nitrate treatments

PubMed Central

Canales, Javier; Moyano, Tomás C.; Villarroel, Eva; Gutiérrez, Rodrigo A.

2014-01-01

Nitrogen (N) is an essential macronutrient for plant growth and development. Plants adapt to changes in N availability partly by changes in global gene expression. We integrated publicly available root microarray data under contrasting nitrate conditions to identify new genes and functions important for adaptive nitrate responses in Arabidopsis thaliana roots. Overall, more than 2000 genes exhibited changes in expression in response to nitrate treatments in Arabidopsis thaliana root organs. Global regulation of gene expression by nitrate depends largely on the experimental context. However, despite significant differences from experiment to experiment in the identity of regulated genes, there is a robust nitrate response of specific biological functions. Integrative gene network analysis uncovered relationships between nitrate-responsive genes and 11 highly co-expressed gene clusters (modules). Four of these gene network modules have robust nitrate responsive functions such as transport, signaling, and metabolism. Network analysis hypothesized G2-like transcription factors are key regulatory factors controlling transport and signaling functions. Our meta-analysis highlights the role of biological processes not studied before in the context of the nitrate response such as root hair development and provides testable hypothesis to advance our understanding of nitrate responses in plants. PMID:24570678

Nitration of naphthalene and remarks on the mechanism of electrophilic aromatic nitration.

PubMed

Olah, G A; Narang, S C; Olah, J A

1981-06-01

Naphthalene was nitrated with a variety of nitrating agents. Comparison of data with Perrin's electrochemical nitration [Perrin, C. L. (1977) J. Am. Chem. Soc. 99, 5516-5518] shows that nitration of naphthalene gives an alpha-nitronaphthalene to beta-nitronaphthalene ratio that varies between 9 and 29 and is thus not constant. Perrin's data, therefore, are considered to be inconclusive evidence for the proposed one-electron transfer mechanism for the nitration of naphthalene and other reactive aromatics. Moodie and Schoefield [Hoggett, J. G., Moodie, R. B., Penton, J. R. & Schoefield, K. (1971) Nitration and Aromatic Reactivity (Cambridge Univ. Press, London)], as well as Perrin, independently concluded that, in the general scheme of nitration of reactive aromatics, there is the necessity to introduce into the classical Ingold mechanism an additional step involving a distinct intermediate preceding the formation of the Wheland intermediate (sigma complexes). This view coincides with our two-step mechanistic picture [Kuhn, S. J. & Olah, G. A. (1961) J. Am. Chem. Soc. 83, 4564-4571] of the nitronium salt nitration of aromatic hydrocarbons (including benzene and toluene), in which low substrate selectivity but high positional selectivity was found, indicating the independence of substrate from positional selectivity.

Nitration of naphthalene and remarks on the mechanism of electrophilic aromatic nitration*

PubMed Central

Olah, George A.; Narang, Subhash C.; Olah, Judith A.

1981-01-01

Naphthalene was nitrated with a variety of nitrating agents. Comparison of data with Perrin's electrochemical nitration [Perrin, C. L. (1977) J. Am. Chem. Soc. 99, 5516-5518] shows that nitration of naphthalene gives an α-nitronaphthalene to β-nitronaphthalene ratio that varies between 9 and 29 and is thus not constant. Perrin's data, therefore, are considered to be inconclusive evidence for the proposed one-electron transfer mechanism for the nitration of naphthalene and other reactive aromatics. Moodie and Schoefield [Hoggett, J. G., Moodie, R. B., Penton, J. R. & Schoefield, K. (1971) Nitration and Aromatic Reactivity (Cambridge Univ. Press, London)], as well as Perrin, independently concluded that, in the general scheme of nitration of reactive aromatics, there is the necessity to introduce into the classical Ingold mechanism an additional step involving a distinct intermediate preceding the formation of the Wheland intermediate (σ complexes). This view coincides with our two-step mechanistic picture [Kuhn, S. J. & Olah, G. A. (1961) J. Am. Chem. Soc. 83, 4564-4571] of the nitronium salt nitration of aromatic hydrocarbons (including benzene and toluene), in which low substrate selectivity but high positional selectivity was found, indicating the independence of substrate from positional selectivity. PMID:16593026

Electrophilic and free radical nitration of benzene and toluene with various nitrating agents*

PubMed Central

Olah, George A.; Lin, Henry C.; Olah, Judith A.; Narang, Subhash C.

1978-01-01

Electrophilic nitration of toluene and benzene was studied under various conditions with several nitrating systems. It was found that high orthopara regioselectivity is prevalent in all reactions and is independent of the reactivity of the nitrating agent. The methyl group of toluene is predominantly ortho-para directing under all reaction conditions. Steric factors are considered to be important but not the sole reason for the variation in the ortho/para ratio. The results reinforce our earlier views that, in electrophilic aromatic nitrations with reactive nitrating agents, substrate and positional selectivities are determined in two separate steps. The first step involves a π-aromatic-NO2+ ion complex or encounter pair, whereas the subsequent step is of arenium ion nature (separate for the ortho, meta, and para positions). The former determines substrate selectivity, whereas the latter determines regioselectivity. Thermal free radical nitration of benzene and toluene with tetranitromethane in sharp contrast gave nearly statistical product distributions. PMID:16592503

Beetroot Juice Supplementation Improves High-Intensity Intermittent Type Exercise Performance in Trained Soccer Players.

PubMed

Nyakayiru, Jean; Jonvik, Kristin L; Trommelen, Jorn; Pinckaers, Philippe J M; Senden, Joan M; van Loon, Luc J C; Verdijk, Lex B

2017-03-22

It has been shown that nitrate supplementation can enhance endurance exercise performance. Recent work suggests that nitrate ingestion can also increase intermittent type exercise performance in recreational athletes. We hypothesized that six days of nitrate supplementation can improve high-intensity intermittent type exercise performance in trained soccer players. Thirty-two male soccer players (age: 23 ± 1 years, height: 181 ± 1 m, weight: 77 ± 1 kg, playing experience: 15.2 ± 0.5 years, playing in the first team of a 2nd or 3rd Dutch amateur league club) participated in this randomized, double-blind cross-over study. All subjects participated in two test days in which high-intensity intermittent running performance was assessed using the Yo-Yo IR1 test. Subjects ingested nitrate-rich (140 mL; ~800 mg nitrate/day; BR) or a nitrate-depleted beetroot juice (PLA) for six subsequent days, with at least eight days of wash-out between trials. The distance covered during the Yo-Yo IR1 was the primary outcome measure, while heart rate (HR) was measured continuously throughout the test, and a single blood and saliva sample were collected just prior to the test. Six days of BR ingestion increased plasma and salivary nitrate and nitrite concentrations in comparison to PLA ( p < 0.001), and enhanced Yo-Yo IR1 test performance by 3.4 ± 1.3% (from 1574 ± 47 to 1623 ± 48 m; p = 0.027). Mean HR was lower in the BR (172 ± 2) vs. PLA trial (175 ± 2; p = 0.014). Six days of BR ingestion effectively improves high-intensity intermittent type exercise performance in trained soccer players.

Beetroot Juice Supplementation Improves High-Intensity Intermittent Type Exercise Performance in Trained Soccer Players

PubMed Central

Nyakayiru, Jean; Jonvik, Kristin L.; Trommelen, Jorn; Pinckaers, Philippe J. M.; Senden, Joan M.; van Loon, Luc J. C.; Verdijk, Lex B.

2017-01-01

It has been shown that nitrate supplementation can enhance endurance exercise performance. Recent work suggests that nitrate ingestion can also increase intermittent type exercise performance in recreational athletes. We hypothesized that six days of nitrate supplementation can improve high-intensity intermittent type exercise performance in trained soccer players. Thirty-two male soccer players (age: 23 ± 1 years, height: 181 ± 1 m, weight: 77 ± 1 kg, playing experience: 15.2 ± 0.5 years, playing in the first team of a 2nd or 3rd Dutch amateur league club) participated in this randomized, double-blind cross-over study. All subjects participated in two test days in which high-intensity intermittent running performance was assessed using the Yo-Yo IR1 test. Subjects ingested nitrate-rich (140 mL; ~800 mg nitrate/day; BR) or a nitrate-depleted beetroot juice (PLA) for six subsequent days, with at least eight days of wash-out between trials. The distance covered during the Yo-Yo IR1 was the primary outcome measure, while heart rate (HR) was measured continuously throughout the test, and a single blood and saliva sample were collected just prior to the test. Six days of BR ingestion increased plasma and salivary nitrate and nitrite concentrations in comparison to PLA (p < 0.001), and enhanced Yo-Yo IR1 test performance by 3.4 ± 1.3% (from 1574 ± 47 to 1623 ± 48 m; p = 0.027). Mean HR was lower in the BR (172 ± 2) vs. PLA trial (175 ± 2; p = 0.014). Six days of BR ingestion effectively improves high-intensity intermittent type exercise performance in trained soccer players. PMID:28327503

Purine uptake in Plasmodium: transport versus metabolism.

PubMed

Kirk, Kiaran; Howitt, Susan M; Bröer, Stefan; Saliba, Kevin J; Downie, Megan J

2009-06-01

In a recent paper, Quashie et al. have proposed that purine uptake into the intraerythrocytic malaria parasite involves four different plasma membrane transporters - two high affinity and two low affinity. They equate one of the two high-affinity transporters with PfNT1, a transporter reported previously to be a low-affinity system. Here, we offer an alternative interpretation of their data, suggesting that the conclusions drawn by Quashie et al. take insufficient account of metabolism.

Host-Guest Complexes with Protein-Ligand-Like Affinities: Computational Analysis and Design

PubMed Central

Moghaddam, Sarvin; Inoue, Yoshihisa

2009-01-01

It has recently been discovered that guests combining a nonpolar core with cationic substituents bind cucurbit[7]uril (CB[7]) in water with ultra-high affinities. The present study uses the Mining Minima algorithm to study the physics of these extraordinary associations and to computationally test a new series of CB[7] ligands designed to bind with similarly high affinity. The calculations reproduce key experimental observations regarding the affinities of ferrocene-based guests with CB[7] and β-cyclodextrin and provide a coherent view of the roles of electrostatics and configurational entropy as determinants of affinity in these systems. The newly designed series of compounds is based on a bicyclo[2.2.2]octane core, which is similar in size and polarity to the ferrocene core of the existing series. Mining Minima predicts that these new compounds will, like the ferrocenes, bind CB[7] with extremely high affinities. PMID:19133781

Structural Insights into the Affinity of Cel7A Carbohydrate-binding Module for Lignin*

PubMed Central

Strobel, Kathryn L.; Pfeiffer, Katherine A.; Blanch, Harvey W.; Clark, Douglas S.

2015-01-01

The high cost of hydrolytic enzymes impedes the commercial production of lignocellulosic biofuels. High enzyme loadings are required in part due to their non-productive adsorption to lignin, a major component of biomass. Despite numerous studies documenting cellulase adsorption to lignin, few attempts have been made to engineer enzymes to reduce lignin binding. In this work, we used alanine-scanning mutagenesis to elucidate the structural basis for the lignin affinity of Trichoderma reesei Cel7A carbohydrate binding module (CBM). T. reesei Cel7A CBM mutants were produced with a Talaromyces emersonii Cel7A catalytic domain and screened for their binding to cellulose and lignin. Mutation of aromatic and polar residues on the planar face of the CBM greatly decreased binding to both cellulose and lignin, supporting the hypothesis that the cellulose-binding face is also responsible for lignin affinity. Cellulose and lignin affinity of the 31 mutants were highly correlated, although several mutants displayed selective reductions in lignin or cellulose affinity. Four mutants with increased cellulose selectivity (Q2A, H4A, V18A, and P30A) did not exhibit improved hydrolysis of cellulose in the presence of lignin. Further reduction in lignin affinity while maintaining a high level of cellulose affinity is thus necessary to generate an enzyme with improved hydrolysis capability. This work provides insights into the structural underpinnings of lignin affinity, identifies residues amenable to mutation without compromising cellulose affinity, and informs engineering strategies for family one CBMs. PMID:26209638

Effect of Nitrates, Thiocyanates and Selenium on the Iron and Iodine Status of Postpartum Women.

PubMed

Bivolarska, Anelia V; Maneva, Ana I; Gatseva, Penka D; Katsarova, Mariana N

2016-09-01

To find correlations between high thiocyanate and nitrate levels and low selenium levels and the indicators of the iodine and iron status of postpartum women. The study included 41 mothers aged 26.4±5.9 yrs from Asenovgrad and nearby villages. Urinary iodine was determined by the Sandell-Kolthoff reaction and thiocyanate - by the interaction of these ions with acidic solution of KMnO4; for serum nitrates we used the colorimetric method; serum selenium was assessed by electro-thermal atomic-absorption spectrophotometry; thyroxin (FT4), the thyroid stimulating hormone (TSH), serum ferritin (SF), and serum transferrin receptor (sTfR) were determined using ELISA; Hb levels were determined by hematology analyzer. Assessing the iodine status, we found a negative correlation between the levels of iodine and thiocyanates in urine (R=-0.717, р<0.0001), a positive correlation between nitrates and TSH (R=0.487, р=0.003) and a negative correlation between nitrates and FT4 (R=-0.312, р=0.06). For the iron status, we found a negative correlation between nitrates and SF (R=-0.429, р=0.009) and between nitrates and Hb (R=-0.383, р=0.021). The Mann-Whitney U-test showed that in women with nitrate levels higher than the mean value there was low FT4 level (р=0.06), high TSH level (р=0.013), low Hb concentration (р=0.061) and low SF concentration (р=0.005). The combined effects of environmental factors (elevated nitrate levels and low selenium level) on the iodine and iron status are manifested by low concentrations of FT4 (р=0.033), Hb (р=0.06) and SF (р=0.05) and high level of TSH (р=0.05). In conclusion, we found that environmental factors, especially when combined, have a negative impact on the iron and iodine status of females.

Predicting nitrate discharge dynamics in mesoscale catchments using the lumped StreamGEM model and Bayesian parameter inference

NASA Astrophysics Data System (ADS)

Woodward, Simon James Roy; Wöhling, Thomas; Rode, Michael; Stenger, Roland

2017-09-01

The common practice of infrequent (e.g., monthly) stream water quality sampling for state of the environment monitoring may, when combined with high resolution stream flow data, provide sufficient information to accurately characterise the dominant nutrient transfer pathways and predict annual catchment yields. In the proposed approach, we use the spatially lumped catchment model StreamGEM to predict daily stream flow and nitrate concentration (mg L-1 NO3-N) in four contrasting mesoscale headwater catchments based on four years of daily rainfall, potential evapotranspiration, and stream flow measurements, and monthly or daily nitrate concentrations. Posterior model parameter distributions were estimated using the Markov Chain Monte Carlo sampling code DREAMZS and a log-likelihood function assuming heteroscedastic, t-distributed residuals. Despite high uncertainty in some model parameters, the flow and nitrate calibration data was well reproduced across all catchments (Nash-Sutcliffe efficiency against Log transformed data, NSL, in the range 0.62-0.83 for daily flow and 0.17-0.88 for nitrate concentration). The slight increase in the size of the residuals for a separate validation period was considered acceptable (NSL in the range 0.60-0.89 for daily flow and 0.10-0.74 for nitrate concentration, excluding one data set with limited validation data). Proportions of flow and nitrate discharge attributed to near-surface, fast seasonal groundwater and slow deeper groundwater were consistent with expectations based on catchment geology. The results for the Weida Stream in Thuringia, Germany, using monthly as opposed to daily nitrate data were, for all intents and purposes, identical, suggesting that four years of monthly nitrate sampling provides sufficient information for calibration of the StreamGEM model and prediction of catchment dynamics. This study highlights the remarkable effectiveness of process based, spatially lumped modelling with commonly available monthly stream sample data, to elucidate high resolution catchment function, when appropriate calibration methods are used that correctly handle the inherent uncertainties.

Sources of nitrate yields in the Mississippi River Basin.

PubMed

David, Mark B; Drinkwater, Laurie E; McIsaac, Gregory F

2010-01-01

Riverine nitrate N in the Mississippi River leads to hypoxia in the Gulf of Mexico. Several recent modeling studies estimated major N inputs and suggested source areas that could be targeted for conservation programs. We conducted a similar analysis with more recent and extensive data that demonstrates the importance of hydrology in controlling the percentage of net N inputs (NNI) exported by rivers. The average fraction of annual riverine nitrate N export/NNI ranged from 0.05 for the lower Mississippi subbasin to 0.3 for the upper Mississippi River basin and as high as 1.4 (4.2 in a wet year) for the Embarras River watershed, a mostly tile-drained basin. Intensive corn (Zea mays L.) and soybean [Glycine max (L.) Merr.] watersheds on Mollisols had low NNI values and when combined with riverine N losses suggest a net depletion of soil organic N. We used county-level data to develop a nonlinear model ofN inputs and landscape factors that were related to winter-spring riverine nitrate yields for 153 watersheds within the basin. We found that river runoff times fertilizer N input was the major predictive term, explaining 76% of the variation in the model. Fertilizer inputs were highly correlated with fraction of land area in row crops. Tile drainage explained 17% of the spatial variation in winter-spring nitrate yield, whereas human consumption of N (i.e., sewage effluent) accounted for 7%. Net N inputs were not a good predictor of riverine nitrate N yields, nor were other N balances. We used this model to predict the expected nitrate N yield from each county in the Mississippi River basin; the greatest nitrate N yields corresponded to the highly productive, tile-drained cornbelt from southwest Minnesota across Iowa, Illinois, Indiana, and Ohio. This analysis can be used to guide decisions about where efforts to reduce nitrate N losses can be most effectively targeted to improve local water quality and reduce export to the Gulf of Mexico.

User’s Guide and History of ANFO (Ammonium Nitrate/Fuel Oil) as a Nuclear Weapons Effect Simulation Explosive

DTIC Science & Technology

1983-03-31

SHOCK SIMULATION 1659 - Amonium nitrate first prepared by Glauber 1867 - Swedish patent granted to Ohlsson and Norrbein for use of ammonium nitrate ...neceessay aqd identify by block number) Ammonium Nitrate -Fuel Oil Aiiblast - . ANFO . Craters High Explosives Explosive Charge Construction * Nuclear...utilizatilon of ANFO for future W FJOAMw. 1473- EDIT00 or INOW ,Sois 0"LTZ"" DO t 473 UNCLASSIFIED SECUM"TY CLASSIFfCATIOl# OF THIS PAGEI(Whonf D Ia LI L

Factors influencing protein tyrosine nitration--structure-based predictive models.

PubMed

Bayden, Alexander S; Yakovlev, Vasily A; Graves, Paul R; Mikkelsen, Ross B; Kellogg, Glen E

2011-03-15

Models for exploring tyrosine nitration in proteins have been created based on 3D structural features of 20 proteins for which high-resolution X-ray crystallographic or NMR data are available and for which nitration of 35 total tyrosines has been experimentally proven under oxidative stress. Factors suggested in previous work to enhance nitration were examined with quantitative structural descriptors. The role of neighboring acidic and basic residues is complex: for the majority of tyrosines that are nitrated the distance to the heteroatom of the closest charged side chain corresponds to the distance needed for suspected nitrating species to form hydrogen bond bridges between the tyrosine and that charged amino acid. This suggests that such bridges play a very important role in tyrosine nitration. Nitration is generally hindered for tyrosines that are buried and for those tyrosines for which there is insufficient space for the nitro group. For in vitro nitration, closed environments with nearby heteroatoms or unsaturated centers that can stabilize radicals are somewhat favored. Four quantitative structure-based models, depending on the conditions of nitration, have been developed for predicting site-specific tyrosine nitration. The best model, relevant for both in vitro and in vivo cases, predicts 30 of 35 tyrosine nitrations (positive predictive value) and has a sensitivity of 60/71 (11 false positives). Copyright © 2010 Elsevier Inc. All rights reserved.

Molybdenum Availability Is Key to Nitrate Removal in Contaminated Groundwater Environments

PubMed Central

Thorgersen, Michael P.; Lancaster, W. Andrew; Vaccaro, Brian J.; Poole, Farris L.; Rocha, Andrea M.; Mehlhorn, Tonia; Pettenato, Angelica; Ray, Jayashree; Waters, R. Jordan; Melnyk, Ryan A.; Chakraborty, Romy; Deutschbauer, Adam M.; Arkin, Adam P.

2015-01-01

The concentrations of molybdenum (Mo) and 25 other metals were measured in groundwater samples from 80 wells on the Oak Ridge Reservation (ORR) (Oak Ridge, TN), many of which are contaminated with nitrate, as well as uranium and various other metals. The concentrations of nitrate and uranium were in the ranges of 0.1 μM to 230 mM and <0.2 nM to 580 μM, respectively. Almost all metals examined had significantly greater median concentrations in a subset of wells that were highly contaminated with uranium (≥126 nM). They included cadmium, manganese, and cobalt, which were 1,300- to 2,700-fold higher. A notable exception, however, was Mo, which had a lower median concentration in the uranium-contaminated wells. This is significant, because Mo is essential in the dissimilatory nitrate reduction branch of the global nitrogen cycle. It is required at the catalytic site of nitrate reductase, the enzyme that reduces nitrate to nitrite. Moreover, more than 85% of the groundwater samples contained less than 10 nM Mo, whereas concentrations of 10 to 100 nM Mo were required for efficient growth by nitrate reduction for two Pseudomonas strains isolated from ORR wells and by a model denitrifier, Pseudomonas stutzeri RCH2. Higher concentrations of Mo tended to inhibit the growth of these strains due to the accumulation of toxic concentrations of nitrite, and this effect was exacerbated at high nitrate concentrations. The relevance of these results to a Mo-based nitrate removal strategy and the potential community-driving role that Mo plays in contaminated environments are discussed. PMID:25979890

Occurrence of nitrate in Tanzanian groundwater aquifers: A review

NASA Astrophysics Data System (ADS)

Elisante, Eliapenda; Muzuka, Alfred N. N.

2017-03-01

More than 25 % of Tanzanian depends on groundwater as the main source of water for drinking, irrigation and industrial activities. The current trend of land use may lead to groundwater contamination and thus increasing risks associated with the usage of contaminated water. Nitrate is one of the contaminants resulting largely from anthropogenic activities that may find its way to the aquifers and thus threatening the quality of groundwater. Elevated levels of nitrate in groundwater may lead to human health and environmental problems. The current trend of land use in Tanzania associated with high population growth, poor sanitation facilities and fertilizer usage may lead to nitrate contamination of groundwater. This paper therefore aimed at providing an overview of to what extent human activities have altered the concentration of nitrate in groundwater aquifers in Tanzania. The concentration of nitrate in Tanzanian groundwater is variable with highest values observable in Dar es Salaam (up to 477.6 mg/l), Dodoma (up to 441.1 mg/l), Tanga (above 100 mg/l) and Manyara (180 mg/l). Such high values can be attributed to various human activities including onsite sanitation in urban centres and agricultural activities in rural areas. Furthermore, there are some signs of increasing concentration of nitrate in groundwater with time in some areas in response to increased human activities. However, reports on levels and trends of nitrate in groundwater in many regions of the country are lacking. For Tanzania to appropriately address the issue of groundwater contamination, a deliberate move to determine nitrate concentration in groundwater is required, as well as protection of recharge basins and improvement of onsite sanitation systems.

Chemical generation of nitric oxide in the mouth from the enterosalivary circulation of dietary nitrate.

PubMed

Duncan, C; Dougall, H; Johnston, P; Green, S; Brogan, R; Leifert, C; Smith, L; Golden, M; Benjamin, N

1995-06-01

High concentrations of nitrite present in saliva (derived from dietary nitrate) may, upon acidification, generate nitrogen oxides in the stomach in sufficient amounts to provide protection from swallowed pathogens. We now show that, in the rat, reduction of nitrate to nitrite is confined to a specialized area on the posterior surface of the tongue, which is heavily colonized by bacteria, and that nitrate reduction is absent in germ-free rats. We also show that in humans increased salivary nitrite production resulting from nitrate intake enhances oral nitric oxide production. We propose that the salivary generation of nitrite is accomplished by a symbiotic relationship involving nitrate-reducing bacteria on the tongue surface, which is designed to provide host defence against microbial pathogens in the mouth and lower gut. These results provide further evidence for beneficial effects of dietary nitrate.

High-affinity K+ uptake in pepper plants.

PubMed

Martínez-Cordero, M Angeles; Martínez, Vicente; Rubio, Francisco

2005-06-01

High-affinity K+ uptake is an essential process for plant nutrition under K+-limiting conditions. The results presented here demonstrate that pepper (Capsicum annuum) plants grown in the absence of NH4+ and starved of K+ show an NH4+-sensitive high-affinity K+ uptake that allows plant roots to deplete external K+ to values below 1 microM. When plants are grown in the presence of NH4+, high-affinity K+ uptake is not inhibited by NH4+. Although NH4+-grown plants deplete external K+ below 1 microM in the absence of NH4+, when 1 mM NH4+ is present they do not deplete external K+ below 10 microM. A K+ transporter of the HAK family, CaHAK1, is very likely mediating the NH4+-sensitive component of the high-affinity K+ uptake in pepper roots. CaHAK1 is strongly induced in the roots that show the NH4+-sensitive high-affinity K+ uptake and its induction is reduced in K+-starved plants grown in the presence of NH4+. The NH4+-insensitive K+ uptake may be mediated by an AKT1-like K+ channel.

High l-Trp affinity of indoleamine 2,3-dioxygenase 1 is attributed to two residues located in the distal heme pocket.

PubMed

Yuasa, Hajime J

2016-10-01

Indoleamine 2, 3-dioxygenase (IDO) catalyzes the oxidative cleavage of the pyrrole ring of l-Trp to generate N-formyl-kynurenine. Two IDO genes, IDO1 and IDO2, are found in vertebrates. Mammalian IDO1s are high-affinity, l-Trp-degrading enzymes, whereas IDO2s generally have a relatively low affinity. It has been suggested that the distal-Ser (corresponding to Ser167 of human IDO1) was crucial for improvement in the affinity for l-Trp but this idea was insufficient to explain the high affinity shown by mammalian IDO1. In this study, the amino acid sequences of vertebrate ancestral IDO1 and ancestral IDO2 were inferred, and bacterially expressed ancestral IDOs were characterized. Although the amino acid sequences of the enzymes shared high identity (86%) with each other, they showed distinct enzymatic properties. In analyses of a series of ancestral IDO1/IDO2 chimeric enzymes and their variants, the distal-Tyr (corresponding to Tyr126 of human IDO1) was detected as another and was probably the most crucial residue for high l-Trp affinity. The two amino acid substitutions (distal-Ser to Thr and distal-Tyr to His) drastically decreased the l-Trp affinity and catalytic efficiency of IDO1s. Conversely, two substitutions (distal-Thr to Ser and distal-His to Tyr) were sufficient to bestow IDO1-like high affinity on ancestral and chicken IDO2. © 2016 Federation of European Biochemical Societies.

Sensitivity to nitrate and nitrite in pond-breeding amphibians from the Pacific Northwest, USA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marco, A.; Quilchano, C.; Blaustein, A.R.

1999-12-01

In static experiments, the authors studied the effects of nitrate and nitrate solutions on newly hatched larvae of five species of amphibians, namely Rana pretiosa, Rana aurora, Bufo boreas, Hyla regilla, and Ambystoma gracile. When nitrate or nitrite ions were added to the water, some larvae of some species reduced feeding activity, swam less vigorously, showed disequilibrium and paralysis, suffered abnormalities and edemas, and eventually died. The observed effects increased with both concentration and time, and there were significant differences in sensitivity among species. Ambrystoma gracile displayed the highest acute effect in water with nitrate and nitrite. The three ranidmore » species had acute effects in water with nitrite. In chronic exposures, R. pretiosa was the most sensitive species to nitrates and nitrites. All species showed 15-d LC50s lower than 2 mg N-NO{sub 2{sup {minus}}}/L. For both N ions, B. boreas was the least sensitive amphibian. All species showed a high morality at the US Environmental Protection Agency-recommended limits of nitrite for warm-water fishes and a significant larval mortality at the recommended limits of nitrite concentration for drinking water. The recommended levels of nitrate for warm-water fishes were highly toxic for R. pretiosa and A. gracile larvae.« less

DOC and nitrate export linked to dominant rainfall-runoff processes, end-members and seasonality - a long-term high frequency measurement campaign

NASA Astrophysics Data System (ADS)

Schwab, M. P.; Klaus, J.; Pfister, L.; Weiler, M.

2016-12-01

Over the past decades, stream sampling protocols for hydro-geochemical parameters were often limited by logistical and technological constraints. While long-term monitoring protocols were typically based on weekly sampling intervals, high frequency sampling was commonly limited to a few single events. In this contribution, we combined high frequency and long-term measurements to understand DOC and nitrate dynamics in a forest headwater for different runoff events and seasons. Our study area is the forested Weierbach catchment (0.47 km2) in Luxembourg, where the fractured schist bedrock is covered by cambisol soils. The runoff response is characterized by a double peak behaviour. The first peak occurs during or right after a rainfall event triggered by fast near surface runoff generation processes, while a second delayed peak lasts several days and is generated by subsurface flow. This second peak occurs only if a distinct storage threshold of the catchment is exceeded. Our observations were carried out with a field deployable UV-Vis spectrometer measuring DOC and nitrate concentrations in-situ at 15 min intervals for more than two years. In addition, a long-term validation was carried out with data obtained from the analysis of water collected with grab samples. The long-term, high-frequency measurements allowed us to calculate a complete and detailed balance of DOC and nitrate export over two years. Transport behaviour of the DOC and nitrate showed different dynamics between the first and second hydrograph peaks. DOC is mainly exported during the first peaks, while nitrate is mostly exported during the delayed second peaks. Biweekly end-member measurement of soil and groundwater over several years enables us to link the behaviour of DOC and nitrate export to various end-members in the catchment. Altogether, the long-term and high-frequency time series provides the opportunity to study DOC and nitrate export processes without having to just rely only on either a few single event measurements or coarse measurement protocols.

Identifying the Source of High-Nitrate Ground Water Related to Artificial Recharge in a Desert Basin

NASA Astrophysics Data System (ADS)

Densmore, J. N.; Nishikawa, T.; Bohlke, J. K.; Martin, P.

2002-12-01

Ground water has been the sole source of water supply for the community of Yucca Valley in the Mojave Desert, California. Domestic wastewater from the community is treated using septic tanks. An imbalance between ground-water recharge and pumpage caused ground-water levels in the ground-water basin to decline by as much as 300 feet from the late 1940s through 1994. In response to this decline, the local water district, Hi-Desert Water District, began an artificial recharge program in February 1995 to replenish the ground water in the basin using imported surface water. The artificial recharge program resulted in water-level recovery of about 250 feet between February 1995 and December 2001; however, nitrate concentrations in some wells also increased from a background concentration of 10 mg/L as NO3 to more than the U.S. Environmental Protection Agency maximum contaminant level of 45 mg/L as NO3, limiting water use for some wells. The largest increase in nitrate concentrations occurred adjacent to the artificial recharge sites where the largest increase in water levels occurred even though the recharge water had low nitrate concentrations. The source of high nitrate concentrations observed in ground water during aquifer recovery was identified by compiling historical water-quality data; monitoring changes in water quality since artificial recharge began; and analyzing selected samples for major-ion chemistry, stable isotopes of H,O, and N, caffeine, and pharmaceuticals. The major-ions and H and O stable-isotope data indicate that ground-water samples that had the highest nitrate concentrations were mixtures of imported water and native ground water. Nitrate-to-chloride ratios, N isotopes and caffeine and pharmaceutical data indicate septic-tank seepage (septage) is the primary source of increases in nitrate concentration. The rapid rise in water levels entrained the large volume of high-nitrate septage stored in the unsaturated zone, resulting in the rapid increase in nitrate concentrations. Results of this study indicate that the potential for ground-water contamination should be evaluated before beginning an artificial recharge program in an area that uses septic tanks.

Mining nitrate concentration patterns from high-frequency in situ monitoring: a step towards more detailed understanding of hydrological processes?

NASA Astrophysics Data System (ADS)

Aubert, Alice; Houska, Tobias; Plesca, Ina; Kraft, Philipp; Breuer, Lutz

2015-04-01

Recently developed sensing technics allow collecting a considerable amount of high-frequency data; not only for hydrologic parameters (water levels, rainfall, etc.) but also for water chemistry. With devices such as in situ spectrophotometer, nitrate concentration can be monitored down to sub-hourly intervals. Thus, opening the way to new questions: what about daily or sub-daily instream nitrate concentration variations? What do these newly observed variations tell us about hydrological processes? In the Vollnkirchener Bach catchment, a headwater creek flows through a human impacted landscape dominated by agricultural and forest use and including a small settlement. Since March 2013, a Pro-PS device has been installed at the gauging station (monitored since 2011). Nitrate concentration is measured every 15 minutes, discharge and water temperature every 5 minutes. Data mining, more precisely motif discovery, is performed on these time series to identify high-resolution patterns. Spectral analysis highlighted that, in data measured at sub-hourly sampling frequency, variations up to a few hours are more likely to be dominated by measurement noise rather than real-world fluctuations. Therefore, we focus on daily motifs and flood patterns (given the fact that hydrological conditions are changing during flood events, we assume that nitrate concentration changes are depicting real processes). Various flood motifs were extracted: (1) nitrate can either be diluted or (2) concentrated, or (3) both (dilution followed by a bumpy recession curve indicating nitrate enrichment at the end of the flood). In addition to these classical nutrient-discharge behaviors, a variety of other interesting motifs were highlighted. (4) A daily nitrate cycle is clearly observed, but only during a specific year period. (5) Lag to peak time between parameters differentiate flood patterns: sometimes nitrate peaks first, sometimes discharge peaks first. (6) Furthermore, we are able to pinpoint the contributions of a combined sewer overflow, as it creates a different motif from diffuse nitrate inflows from adjacent agricultural fields. We look into the other hydrological parameters to explain this variety of patterns and their occurrence time.

Design and conduct of 'Xtreme Alps': a double-blind, randomised controlled study of the effects of dietary nitrate supplementation on acclimatisation to high altitude.

PubMed

Martin, Daniel S; Gilbert-Kawai, Edward T; Meale, Paula M; Fernandez, Bernadette O; Cobb, Alexandra; Khosravi, Maryam; Mitchell, Kay; Grocott, Michael P W; Levett, Denny Z H; Mythen, Michael G; Feelisch, Martin

2013-11-01

The study of healthy human volunteers ascending to high altitude provides a robust model of the complex physiological interplay that emulates human adaptation to hypoxaemia in clinical conditions. Nitric oxide (NO) metabolism may play an important role in both adaptation to high altitude and response to hypoxaemia during critical illness at sea level. Circulating nitrate and nitrite concentrations can be augmented by dietary supplementation and this is associated with improved exercise performance and mitochondrial efficiency. We hypothesised that the administration of a dietary substance (beetroot juice) rich in nitrate would improve oxygen efficiency during exercise at high altitude by enhancing tissue microcirculatory blood flow and oxygenation. Furthermore, nitrate supplementation would lead to measurable increases in NO bioactivity throughout the body. This methodological manuscript describes the design and conduct of the 'Xtreme Alps' expedition, a double-blind randomised controlled trial investigating the effects of dietary nitrate supplementation on acclimatisation to hypobaric hypoxia at high altitude in healthy human volunteers. The primary outcome measure was the change in oxygen efficiency during exercise at high altitude between participants allocated to receive nitrate supplementation and those receiving a placebo. A number of secondary measures were recorded, including exercise capacity, peripheral and microcirculatory blood flow and tissue oxygenation. Results from this study will further elucidate the role of NO in adaption to hypoxaemia and guide clinical trials in critically ill patients. Improved understanding of hypoxaemia in critical illness may provide new therapeutic avenues for interventions that will improve survival in critically ill patients. © 2013. Published by Elsevier Inc. All rights reserved.

Relations that affect the probability and prediction of nitrate concentration in private wells in the glacial aquifer system in the United States

USGS Publications Warehouse

Warner, Kelly L.; Arnold, Terri L.

2010-01-01

Nitrate in private wells in the glacial aquifer system is a concern for an estimated 17 million people using private wells because of the proximity of many private wells to nitrogen sources. Yet, less than 5 percent of private wells sampled in this study contained nitrate in concentrations that exceeded the U.S. Environmental Protection Agency (USEPA) Maximum Contaminant Level (MCL) of 10 mg/L (milligrams per liter) as N (nitrogen). However, this small group with nitrate concentrations above the USEPA MCL includes some of the highest nitrate concentrations detected in groundwater from private wells (77 mg/L). Median nitrate concentration measured in groundwater from private wells in the glacial aquifer system (0.11 mg/L as N) is lower than that in water from other unconsolidated aquifers and is not strongly related to surface sources of nitrate. Background concentration of nitrate is less than 1 mg/L as N. Although overall nitrate concentration in private wells was low relative to the MCL, concentrations were highly variable over short distances and at various depths below land surface. Groundwater from wells in the glacial aquifer system at all depths was a mixture of old and young water. Oxidation and reduction potential changes with depth and groundwater age were important influences on nitrate concentrations in private wells. A series of 10 logistic regression models was developed to estimate the probability of nitrate concentration above various thresholds. The threshold concentration (1 to 10 mg/L) affected the number of variables in the model. Fewer explanatory variables are needed to predict nitrate at higher threshold concentrations. The variables that were identified as significant predictors for nitrate concentration above 4 mg/L as N included well characteristics such as open-interval diameter, open-interval length, and depth to top of open interval. Environmental variables in the models were mean percent silt in soil, soil type, and mean depth to saturated soil. The 10-year mean (1992-2001) application rate of nitrogen fertilizer applied to farms was included as the potential source variable. A linear regression model also was developed to predict mean nitrate concentrations in well networks. The model is based on network averages because nitrate concentrations are highly variable over short distances. Using values for each of the predictor variables averaged by network (network mean value) from the logistic regression models, the linear regression model developed in this study predicted the mean nitrate concentration in well networks with a 95 percent confidence in predictions.

Nitrate Contamination in the groundwater of the Lake Acıgöl Basin, SW Turkey

NASA Astrophysics Data System (ADS)

Karaman, Muhittin; Budakoǧlu, Murat; Taşdelen, Suat

2017-04-01

The lacustrine Acıgöl basin, formed as an extensional half-graben, hosts various bodies of water, such as cold-hot springs, lakes, streams, and wells. The hydrologically closed basin contains a hypersaline lake (Lake Acıgöl) located in the southern part of the basin. The brackish springs and deep waters discharged along the Acıgöl fault zone in the southern part of the basin feed the hypersaline lake. Groundwater is used as drinking, irrigation, and domestic water in the closed Acıgöl Basin. Groundwater flows into the hypersaline lake from the highland. The Acıgöl basin hosts large plains (Hambat, Başmakçı, and Evciler). Waters in agricultural areas contain high amounts of nitrate; groundwater samples in agricultural areas contain nitrate levels higher than 10 mg/L. Nitrate concentrations in the groundwater samples varied from 0 to 487 mg/L (n=165); 25.4 % of the groundwater samples from the basin had nitrate concentrations above 10 mg/L (the WHO drinking guideline) and 52.2% of the groundwater samples from the basin had nitrate concentrations above 3.0 mg/L, and these high values were regarded as the result of human activity. The highest nitrate values were measured in the Hambat plain (480 and 100 mg/L) and Yirce Pinari spring (447 mg/L), which discharges along the Acıgöl fault zone in the southern part of the basin. The average multi-temporal nitrate concentration of the Yirce Pınarı spring was 3.3 mg/L. Extreme nitrate values were measured in the Yirce Pınarı spring during periods when sheep wool was washed (human activity). The lowest nitrate concentrations were observed in some springs that discharged along the Acıgöl fault zone in the southern part of the basin. Nitrate was not detected in deep groundwater discharged along the Acıgöl fault zone. Nitrate concentrations in deep groundwater and some springs discharged along the Acıgöl fault zone and those feeding the hypersaline lake were significantly affected by redox conditions. Nitrate in these reducing waters was transformed into ammonium. Nitrate concentrations in the Acıgöl Basin were enriched in groundwater beneath agricultural areas and this affected redox conditions. The main source of nitrate contamination was agricultural fertilizers. Elevated nitrate concentrations in groundwater, especially in agricultural areas of the Acigol Basin, can cause public health problems and environmental pollution.

Dietary intake of polyphenols, nitrate and nitrite and gastric cancer risk in Mexico City

PubMed Central

Hernández-Ramírez, Raúl U.; Galván-Portillo, Marcia V.; Ward, Mary H.; Agudo, Antonio; González, Carlos A.; Oñate-Ocaña, Luis F.; Herrera-Goepfert, Roberto; Palma-Coca, Oswaldo; López-Carrillo, Lizbeth

2009-01-01

N-Nitroso compounds (NOC) are potent animal carcinogens and potential human carcinogens. The primary source of exposure for most individuals may be endogenous formation, a process that can be inhibited by dietary polyphenols. To estimate the risk of gastric cancer (GC) in relation to the individual and combined consumption of polyphenols and NOC precursors (nitrate and nitrite), a population-based case–control study was carried out in Mexico City from 2004 to 2005 including 257 histologically confirmed GC cases and 478 controls. Intake of polyphenols, nitrate and nitrite were estimated using a food frequency questionnaire. High intakes of cinnamic acids, secoisolariciresinol and coumestrol were associated with an ~50% reduction in GC risk. A high intake of total nitrite as well as nitrate and nitrite from animal sources doubled the GC risk. Odds ratios around 2-fold were observed among individuals with both low intake of cinnamic acids, secoisolariciresinol or coumestrol and high intake of animal-derived nitrate or nitrite, compared to high intake of the polyphenols and low animal nitrate or nitrite intake, respectively. Results were similar for both the intestinal and diffuse types of GC. Our results show, for the first time, a protective effect for GC because of higher intake of cinnamic acids, secoisolariciresinol and coumestrol, and suggest that these polyphenols reduce GC risk through inhibition of endogenous nitrosation. The main sources of these polyphenols were pears, mangos and beans for cinnamic acids; beans, carrots and squash for secoisolariciresinol and legumes for coumestrol. PMID:19449378

Modeled nitrate levels in well water supplies and prevalence of abnormal thyroid conditions among the Old Order Amish in Pennsylvania

PubMed Central

2012-01-01

Background Nitrate is a widespread contaminant of drinking water supplies, especially in agricultural areas. Nitrate intake from drinking water and dietary sources can interfere with the uptake of iodide by the thyroid, thus potentially impacting thyroid function. Methods We assessed the relation of estimated nitrate levels in well water supplies with thyroid health in a cohort of 2,543 Old Order Amish residing in Lancaster, Chester, and Lebanon counties in Pennsylvania for whom thyroid stimulating hormone (TSH) levels were measured during 1995-2008. Nitrate measurement data (1976-2006) for 3,613 wells in the study area were obtained from the U.S. Geological Survey and we used these data to estimate concentrations at study participants' residences using a standard linear mixed effects model that included hydrogeological covariates and kriging of the wells' residuals. Nitrate levels estimated by the model ranged from 0.35 mg/L to 16.4 mg/L N-NO3-, with a median value of 6.5 mg/L, which was used as the cutpoint to define high and low nitrate exposure. In a validation analysis of the model, we calculated that the sensitivity of the model was 67% and the specificity was 93%. TSH levels were used to define the following outcomes: clinical hyperthyroidism (n = 10), clinical hypothyroidism (n = 56), subclinical hyperthyroidism (n = 25), and subclinical hypothyroidism (n = 228). Results In women, high nitrate exposure was significantly associated with subclinical hypothyroidism (OR = 1.60; 95% CI: 1.11-2.32). Nitrate was not associated with subclinical thyroid disease in men or with clinical thyroid disease in men or women. Conclusions Although these data do not provide strong support for an association between nitrate in drinking water and thyroid health, our results do suggest that further exploration of this hypothesis is warranted using studies that incorporate individual measures of both dietary and drinking water nitrate intake. PMID:22339761

Modeled nitrate levels in well water supplies and prevalence of abnormal thyroid conditions among the Old Order Amish in Pennsylvania.

PubMed

Aschebrook-Kilfoy, Briseis; Heltshe, Sonya L; Nuckols, John R; Sabra, Mona M; Shuldiner, Alan R; Mitchell, Braxton D; Airola, Matt; Holford, Theodore R; Zhang, Yawei; Ward, Mary H

2012-02-17

Nitrate is a widespread contaminant of drinking water supplies, especially in agricultural areas. Nitrate intake from drinking water and dietary sources can interfere with the uptake of iodide by the thyroid, thus potentially impacting thyroid function. We assessed the relation of estimated nitrate levels in well water supplies with thyroid health in a cohort of 2,543 Old Order Amish residing in Lancaster, Chester, and Lebanon counties in Pennsylvania for whom thyroid stimulating hormone (TSH) levels were measured during 1995-2008. Nitrate measurement data (1976-2006) for 3,613 wells in the study area were obtained from the U.S. Geological Survey and we used these data to estimate concentrations at study participants' residences using a standard linear mixed effects model that included hydrogeological covariates and kriging of the wells' residuals. Nitrate levels estimated by the model ranged from 0.35 mg/L to 16.4 mg/L N-NO3(-), with a median value of 6.5 mg/L, which was used as the cutpoint to define high and low nitrate exposure. In a validation analysis of the model, we calculated that the sensitivity of the model was 67% and the specificity was 93%. TSH levels were used to define the following outcomes: clinical hyperthyroidism (n = 10), clinical hypothyroidism (n = 56), subclinical hyperthyroidism (n = 25), and subclinical hypothyroidism (n = 228). In women, high nitrate exposure was significantly associated with subclinical hypothyroidism (OR = 1.60; 95% CI: 1.11-2.32). Nitrate was not associated with subclinical thyroid disease in men or with clinical thyroid disease in men or women. Although these data do not provide strong support for an association between nitrate in drinking water and thyroid health, our results do suggest that further exploration of this hypothesis is warranted using studies that incorporate individual measures of both dietary and drinking water nitrate intake.

Mortality of nitrate fertiliser workers.

PubMed Central

Al-Dabbagh, S; Forman, D; Bryson, D; Stratton, I; Doll, R

1986-01-01

An epidemiological cohort study was conducted to investigate the mortality patterns among a group of workers engaged in the production of nitrate based fertilisers. This study was designed to test the hypothesis that individuals exposed to high concentrations of nitrates might be at increased risk of developing cancers, particularly gastric cancer. A total of 1327 male workers who had been employed in the production of fertilisers between 1946 and 1981 and who had been occupationally exposed to nitrates for at least one year were followed up until 1 March 1981. In total, 304 deaths were observed in this group and these were compared with expected numbers calculated from mortality rates in the northern region of England, where the factory was located. Analysis was also carried out separately for a subgroup of the cohort who had been heavily exposed to nitrates--that is, working in an environment likely to contain more than 10 mg nitrate/m3 for a year or longer. In neither the entire cohort nor the subgroup was any significant excess observed for all causes of mortality or for mortality from any of five broad categories of cause or from four specific types of cancer. A small excess of lung cancer was noted more than 20 years after first exposure in men heavily exposed for more than 10 years. That men were exposed to high concentrations of nitrate was confirmed by comparing concentrations of nitrates in the saliva of a sample of currently employed men with control men, employed at the same factory but not in fertiliser production. The men exposed to nitrate had substantially raised concentrations of nitrate in their saliva compared with both controls within the industry and with men in the general population and resident nearby. The results of this study therefore weight against the idea that exposure to nitrates in the environment leads to the formation in vivo of material amounts of carcinogens. PMID:3015194

Nitrate dynamics within a stream-lake network through time and space

NASA Astrophysics Data System (ADS)

Loken, L. C.; Crawford, J. T.; Childress, E. S.; Casson, N. J.; Stanley, E. H.

2014-12-01

Nitrate dynamics in streams are governed by biology, hydrology, and geomorphology, and the ability to parse these drivers apart has improved with the development of accurate high-frequency sensors. By combining a stationary Eulerian and a quasi-Lagrangian sensor platform, we investigated the timing of nitrate flushing and identified locations of elevated biogeochemical cycling along a stream-lake network in Northern Wisconsin, USA. Two years of continuous oxygen, carbon dioxide, and discharge measurements were used to compute gross primary production (GPP) and ecosystem respiration (ER) downstream of a wetland reach of Allequash Creek. Metabolic rates and flow patterns were compared with nitrate concentrations measured every 30 minutes using an optical sensor. Additionally, we floated a sensor array from the headwater spring ponds through a heterogeneous stream reach consisting of wetlands, beaver ponds, forested segments, and two lakes. Two distinct temporal patterns of stream nitrate concentrations were observed. During high flow events such as spring snowmelt and summer rain events, nitrate concentrations increased from ~5 μM (baseflow) to 12 μM, suggesting flushing from catchment sources. During baseflow conditions, nitrate followed a diel cycle with a 0.3-1.0 μM daytime draw down. Daily nitrate reduction was positively correlated with GPP calculated from oxygen and carbon dioxide records. Lastly, spatial analyses revealed lowest nitrate concentrations in the wetland reach, approximately 2-3 μM lower than the upstream spring ponds, and downstream lakes and forested reaches. This snapshot implies greater nitrate removal potential in the wetland reach likely driven by denitrification in organic rich sediments and macrophyte uptake in the open canopy stream segment. Taken together the temporal and spatial results show the dynamics of hydrology, geomorphology, and biology to influence nitrate delivery and variability in ecosystem processing through a stream-lake system. Future ecosystem studies could benefit by including multiple reference frameworks to better assess processes not captured by a single station approach.

Pertussis toxin modifies the characteristics of both the inhibitory GTP binding proteins and the somatostatin receptor in anterior pituitary tumor cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mahy, N.; Woolkalis, M.; Thermos, K.

1988-08-01

The effects of pertussis toxin treatment on the characteristics of somatostatin receptors in the anterior pituitary tumor cell line AtT-20 were examined. Pertussis toxin selectively catalyzed the ADP ribosylation of the alpha subunits of the inhibitory GTP binding proteins in AtT-20 cells. Toxin treatment abolished somatostatin inhibition of forskolin-stimulated adenylyl cyclase activity and somatostatin stimulation of GTPase activity. To examine the effects of pertussis toxin treatment on the characteristics of the somatostatin receptor, the receptor was labeled by the somatostatin analog (125I)CGP 23996. (125I)CGP 23996 binding to AtT-20 cell membranes was saturable and within a limited concentration range was tomore » a single high affinity site. Pertussis toxin treatment reduced the apparent density of the high affinity (125I)CGP 23996 binding sites in AtT-20 cell membranes. Inhibition of (125I)CGP 23996 binding by a wide concentration range of CGP 23996 revealed the presence of two binding sites. GTP predominantly reduced the level of high affinity sites in control membranes. Pertussis toxin treatment also diminished the amount of high affinity sites. GTP did not affect (125I)CGP 23996 binding in the pertussis toxin-treated membranes. The high affinity somatostatin receptors were covalently labeled with (125I) CGP 23996 and the photoactivated crosslinking agent n-hydroxysuccinimidyl-4-azidobenzoate. No high affinity somatostatin receptors, covalently bound to (125I)CGP 23996, were detected in the pertussis toxin-treated membranes. These results are most consistent with pertussis toxin uncoupling the inhibitory G proteins from the somatostatin receptor thereby converting the receptor from a mixed population of high and low affinity sites to only low affinity receptors.« less

Absence of an effect of high nitrate intake from beetroot juice on blood pressure in treated hypertensive individuals: a randomized controlled trial.

PubMed

Bondonno, Catherine P; Liu, Alex H; Croft, Kevin D; Ward, Natalie C; Shinde, Sujata; Moodley, Yuben; Lundberg, Jon O; Puddey, Ian B; Woodman, Richard J; Hodgson, Jonathan M

2015-08-01

Dietary nitrate, which is in green leafy vegetables and beetroot, decreases blood pressure through the enterosalivary nitrate-nitrite-nitric oxide pathway in healthy individuals. Whether similar effects would occur in individuals with treated hypertension and, therefore, at increased risk of cardiovascular disease is unclear. We assessed whether increased dietary nitrate intake by using beetroot juice for 1 wk lowers blood pressure in treated hypertensive men and women. Participants (n = 27) were recruited to a randomized, placebo-controlled, double-blind crossover trial. The effect of 1-wk intake of nitrate-rich beetroot juice was compared with 1-wk intake of nitrate-depleted beetroot juice (placebo). The primary outcome was blood pressure assessed by measuring home blood pressure during the intervention and 24-h ambulatory blood pressure on day 7 of the intervention. Other outcomes included nitrate metabolism assessed by measuring nitrate and nitrite in plasma, saliva, and urine. Relative to the placebo, 1-wk intake of nitrate-rich beetroot juice resulted in a 3-fold increase in plasma nitrite and nitrate, a 7-fold increase in salivary nitrite, an 8-fold higher salivary nitrate, and a 4-fold increase in both urinary nitrite and nitrate (P < 0.001). However, no differences in home blood pressure and 24-h ambulatory blood pressure were observed with 1-wk intake of nitrate-rich beetroot juice in comparison with the placebo. An increase in dietary nitrate intake may not be an effective short-term approach to further lower blood pressure in treated hypertensive subjects. © 2015 American Society for Nutrition.

Membrane biological reactors to remove nitrate, digest biosolids, and eliminate water flushing requirements within replicated recirculation systems culturing rainbow trout

USDA-ARS?s Scientific Manuscript database

Nutrients, particularly nitrate (NO3), can accumulate to very high levels within low exchange recirculation aquaculture systems (RAS) and negatively impact a number of cultured species. To prevent the harmful effects of nitrate accumulation and to dispose of concentrated waste biosolids, many RAS ar...

Kinetics of nitrate and perchlorate reduction in ion exchange brine using the membrane biofilm reactor (MBfR)

EPA Science Inventory

Several sources of bacterial inocula were tested for their ability to reduce nitrate and perchlorate in synthetic ion-exchange spent brine (3-4.5% salinity) using a hydrogen-based membrane biofilm reactor (MBfR). Nitrate and perchlorate removal fluxes reached as high as 5.4 g N ...

Variation in benthic metabolism and nitrogen cycling across clam aquaculture sites.

PubMed

Murphy, Anna E; Nizzoli, Daniele; Bartoli, Marco; Smyth, Ashley R; Castaldelli, Giuseppe; Anderson, Iris C

2018-02-01

As bivalve aquaculture expands globally, an understanding of how it alters nitrogen is important to minimize impacts. This study investigated nitrogen cycling associated with clam aquaculture in the Sacca di Goro, Italy (Ruditapes philipinarum) and the Eastern Shore, USA (Mercenaria mercenaria). Ammonium and dissolved oxygen fluxes were positively correlated with clam biomass; R. philippinarum consumed ~6 times more oxygen and excreted ~5 times more NH 4 + than M. mercenaria. There was no direct effect of clams on denitrification or dissimilatory nitrate reduction to ammonium (DNRA); rather, nitrate availability controlled the competition between these microbial pathways. Highest denitrification rates were measured at sites where both water column nitrate and nitrification were elevated due to high densities of a burrowing amphipod (Corophium sp.). DNRA exceeded denitrification where water column nitrate was low and nitrification was suppressed in highly reduced sediment, potentially due to low hydrologic flow and high clam densities. Copyright © 2017 Elsevier Ltd. All rights reserved.

Distinguishing summertime atmospheric production of nitrate across the East Antarctic Ice Sheet

NASA Astrophysics Data System (ADS)

Shi, G.; Buffen, A. M.; Ma, H.; Hu, Z.; Sun, B.; Li, C.; Yu, J.; Ma, T.; An, C.; Jiang, S.; Li, Y.; Hastings, M. G.

2018-06-01

Surface snow and atmospheric samples collected along a traverse from the coast to the ice sheet summit (Dome A) are used to investigate summertime atmospheric production of nitrate (NO3-) across East Antarctica. The strong relationship observed between δ15N and δ18O of nitrate in the surface snow suggests a large (lesser) extent of nitrate photolysis in the interior (coastal) region. A linear correlation between the oxygen isotopes of nitrate (δ18O and Δ17O) indicates mixing of various oxidants that react with NOx (NOx = NO + NO2) to produce atmospheric nitrate. On the plateau, the isotopes of snow nitrate are best explained by local reoxidation chemistry of NOx, possibly occurring in both condensed and gas phases. Nitrate photolysis results in redistribution of snow nitrate, and the plateau snow is a net exporter of nitrate and its precursors. Our results suggest that while snow-sourced NOx from the plateau due to photolysis is a significant input to the nitrate budget in coastal snow (up to ∼35%), tropospheric transport from mid-low latitudes dominates (∼65%) coastal snow nitrate. The linear relationship of δ18O vs. Δ17O of the snow nitrate suggests a predominant role of hydroxyl radical (OH) and ozone (O3) in nitrate production, although a high Δ17O(O3) is required to explain the observations. Across Antarctica the oxygen isotope composition of OH appears to be dominated by exchange with water vapor, despite the very dry environment. One of the largest uncertainties in quantifying nitrate production pathways is the limited knowledge of atmospheric oxidant isotopic compositions.

Cytomegalovirus-Specific CD8+ T-Cells With Different T-Cell Receptor Affinities Segregate T-Cell Phenotypes and Correlate With Chronic Graft-Versus-Host Disease in Patients Post-Hematopoietic Stem Cell Transplantation

PubMed Central

Poiret, Thomas; Axelsson-Robertson, Rebecca; Remberger, Mats; Luo, Xiao-Hua; Rao, Martin; Nagchowdhury, Anurupa; Von Landenberg, Anna; Ernberg, Ingemar; Ringden, Olle; Maeurer, Markus

2018-01-01

Virus-specific T-cell responses are crucial to control cytomegalovirus (CMV) infections/reactivation in immunocompromised individuals. Adoptive cellular therapy with CMV-specific T-cells has become a viable treatment option. High-affinity anti-viral cellular immune responses are associated with improved long-term immune protection against CMV infection. To date, the characterization of high-affinity T-cell responses against CMV has not been achieved in blood from patients after allogeneic hematopoietic stem cell transplantation (HSCT). Therefore, the purpose of this study was to describe and analyze the phenotype and clinical impact of different CMV-specific CD8+ cytotoxic T-lymphocytes (CMV-CTL) classes based on their T-cell receptor (TCR) affinity. T-cells isolated from 23 patients during the first year following HSCT were tested for the expression of memory markers, programmed cell death 1 (PD-1), as well as TCR affinity, using three different HLA-A*02:01 CMVNLVPMVATV-Pp65 tetramers (wild-type, a245v and q226a mutants). High-affinity CMV-CTL defined by q226a tetramer binding, exhibited a higher frequency in CD8+ T-cells in the first month post-HSCT and exhibited an effector memory phenotype associated with strong PD-1 expression as compared to the medium- and low-affinity CMV-CTLs. High-affinity CMV-CTL was found at higher proportion in patients with chronic graft-versus-host disease (p < 0.001). This study provides a first insight into the detailed TCR affinities of CMV-CTL. This may be useful in order to improve current immunotherapy protocols using isolation of viral-specific T-cell populations based on their TCR affinity. PMID:29692783

pMHC affinity controls duration of CD8+ T cell–DC interactions and imprints timing of effector differentiation versus expansion

PubMed Central

Sharpe, James; Zehn, Dietmar; Kreutzfeldt, Mario

2016-01-01

During adaptive immune responses, CD8+ T cells with low TCR affinities are released early into the circulation before high-affinity clones become dominant at later time points. How functional avidity maturation is orchestrated in lymphoid tissue and how low-affinity cells contribute to host protection remains unclear. In this study, we used intravital imaging of reactive lymph nodes (LNs) to show that T cells rapidly attached to dendritic cells irrespective of TCR affinity, whereas one day later, the duration of these stable interactions ceased progressively with lowering peptide major histocompatibility complex (pMHC) affinity. This correlated inversely BATF (basic leucine zipper transcription factor, ATF-like) and IRF4 (interferon-regulated factor 4) induction and timing of effector differentiation, as low affinity–primed T cells acquired cytotoxic activity earlier than high affinity–primed ones. After activation, low-affinity effector CD8+ T cells accumulated at efferent lymphatic vessels for egress, whereas high affinity–stimulated CD8+ T cells moved to interfollicular regions in a CXCR3-dependent manner for sustained pMHC stimulation and prolonged expansion. The early release of low-affinity effector T cells led to rapid target cell elimination outside reactive LNs. Our data provide a model for affinity-dependent spatiotemporal orchestration of CD8+ T cell activation inside LNs leading to functional avidity maturation and uncover a role for low-affinity effector T cells during early microbial containment. PMID:27799622

Fertilizer Induced Nitrate Pollution in RCW: Calibration of the DNDC Model

NASA Astrophysics Data System (ADS)

El Hailouch, E.; Hornberger, G.; Crane, J. W.

2012-12-01

Fertilizer is widely used among urban and suburban households due to the socially driven attention of homeowners to lawn appearance. With high nitrogen content, fertilizer considerably impacts the environment through the emission of the highly potent greenhouse gas nitrous oxide and the leaching of nitrate. Nitrate leaching is significantly important because fertilizer sourced nitrate that is partially leached into soil causes groundwater pollution. In an effort to model the effect of fertilizer application on the environment, the geochemical DeNitrification-DeComposition model (DNDC) was previously developed to quantitatively measure the effects of fertilizer use. The purpose of this study is to use this model more effectively on a large scale through a measurement based calibration. For this reason, leaching was measured and studied on 12 sites in the Richland Creek Watershed (RCW). Information about the fertilization and irrigation regimes of these sites was collected, along with lysimeter readings that gave nitrate fluxes in the soil. A study of the amount and variation in nitrate leaching with respect to the varying geographical locations, time of the year, and fertilization and irrigation regimes has lead to a better understanding of the driving forces behind nitrate leaching. Quantifying the influence of each of these parameters allows for a more accurate calibration of the model thus permitting use that extends beyond the RCW. Measurement of nitrate leaching on a statewide or nationwide level in turn will help guide efforts in the reduction of groundwater pollution caused by fertilizer.

Observations of fine particulate nitrated phenols in four sites in northern China: concentrations, source apportionment, and secondary formation

NASA Astrophysics Data System (ADS)

Wang, Liwei; Wang, Xinfeng; Gu, Rongrong; Wang, Hao; Yao, Lan; Wen, Liang; Zhu, Fanping; Wang, Weihao; Xue, Likun; Yang, Lingxiao; Lu, Keding; Chen, Jianmin; Wang, Tao; Zhang, Yuanghang; Wang, Wenxing

2018-03-01

Filter samples of fine particulate matters were collected at four sites in northern China (urban, rural, and mountain) in summer and winter, and the contents of nine nitrated phenols were quantified in the laboratory with the use of ultra-high-performance liquid chromatography coupled with mass spectrometry. During the sampling periods, the concentrations of particulate nitrated phenols exhibited distinct temporal and spatial variation. On average, the total concentration of particulate nitrated phenols in urban Jinan in the wintertime reached 48.4 ng m-3, and those in the summertime were 9.8, 5.7, 5.9, and 2.5 ng m-3 in urban Jinan, rural Yucheng and Wangdu, and Mt. Tai, respectively. The elevated concentrations of nitrated phenols in wintertime and in urban areas demonstrate the apparent influences of anthropogenic sources. The positive matrix factorization receptor model was then applied to determine the origins of particulate nitrated phenols in northern China. The five major source factors were traffic, coal combustion, biomass burning, secondary formation, and aged coal combustion plume. Among them, coal combustion played a vital role, especially at the urban site in the wintertime, with a contribution of around 55 %. In the summertime, the observed nitrated phenols were highly influenced by aged coal combustion plumes at all of the sampling sites. Meanwhile, in remote areas, contributions from secondary formation were significant. Further correlation analysis indicates that nitrosalicylic acids were produced mostly from secondary formation that was dominated by NO2 nitration.

Colour formation in fermented sausages by meat-associated staphylococci with different nitrite- and nitrate-reductase activities.

PubMed

Gøtterup, Jacob; Olsen, Karsten; Knøchel, Susanne; Tjener, Karsten; Stahnke, Louise H; Møller, Jens K S

2008-04-01

Three Staphylococcus strains, S. carnosus, S. simulans and S. saprophyticus, selected due to their varying nitrite and/or nitrate-reductase activities, were used to initiate colour formation during sausage fermentation. During fermentation of sausages with either nitrite or nitrate added, colour was followed by L(∗)a(∗)b measurements and the content of nitrosylmyoglobin (MbFe(II)NO) quantified by electron spin resonance (ESR). MbFe(II)NO was rapidly formed in sausages with added nitrite independent of the presence of nitrite reducing bacteria, whereas the rate of MbFe(II)NO formation in sausages with added nitrate depended on the specific Staphylococcus strain. Strains with high nitrate-reductase activity showed a significantly faster rate of pigment formation, but other factors were of influence as well. Product stability for the sliced, packaged sausage was evaluated as surface colour and oxidation by autofluorescence and hexanal content, respectively. No significant direct effect of the Staphylococcus addition was observed, however, there was a clear correspondence between high initial amount of MbFe(II)NO in the different sausages and the colour stability during storage. Autofluorescence data correlated well with hexanal content, and may be used as predictive tools. Overall, nitrite- and nitrate-reductase activities of Staphylococcus strains in nitrite-cured sausages were of limited importance regarding colour development, while in nitrate-cured sausages strains with higher nitrate reductase activity were crucial for ensuring optimal colour formation during initial fermentation stages.

Root-type-specific plasticity in response to localized high nitrate supply in maize (Zea mays)

PubMed Central

Yu, Peng; Hochholdinger, Frank; Li, Chunjian

2015-01-01

Background and Aims Shoot-borne roots contribute to most of the nutrient uptake throughout the life cycle of maize (Zea mays). Compared with numerous studies with embryonic roots, detailed information on the phenotypic plasticity of shoot-borne roots in response to a heterogeneous nitrogen supply is scarce. The present study therefore provides a comprehensive profile of fine-scale plastic responses of distinct root types to localized high nitrate supply. Methods Seedlings of the maize inbred line B73 were grown in split-root systems. The anatomy and morphological plasticity of the primary root and the roots initiated from the 2nd, 5th and 7th shoot nodes, and their lateral roots, were studied in response to local high nitrate supply to one side of the root system. Key Results In contrast to the insensitivity of axial roots, local high nitrate supply increased the length of 1st-order lateral roots on the primary root and the three whorls of shoot-borne roots at different growth stages, and increased the density of 1st-order lateral roots on the 7th shoot-borne root after silking. The length and density of 2nd-order lateral roots on the three whorls of shoot-borne roots displayed a more flexible response to local high nitrate than 1st-order lateral roots. Root diameter and number, and total area and diameter of metaxylem vessels increased from the primary root to early and then later developed shoot-borne roots, which showed a positive relationship with shoot growth and N accumulation. Conclusions Maize axial roots and lateral roots responded differently to local high nitrate, and this was related to their function. The extent of morphological plasticity of lateral roots in response to local high nitrate depended on the initiation time of the shoot-borne roots on which the lateral roots developed. Morphological plasticity was higher on 2nd-order than on 1st-order lateral roots. The results suggest that higher order lateral root branching might be a potential target for genetic improvement in future maize breeding. PMID:26346717

Anion binding in the C3v-symmetric cavity of a protonated tripodal amine receptor: potentiometric and single crystal X-ray studies.

PubMed

Bose, Purnandhu; Ravikumar, I; Ghosh, Pradyut

2011-11-07

Tris(2-aminoethyl)amine (tren) based pentafluorophenyl-substituted tripodal L, tris[[(2,3,4,5,6-pentafluorobenzyl)amino]ethyl]amine receptor is synthesized in good yield and characterized by single crystal X-ray diffraction analysis. Detailed structural aspects of binding of different anionic guests toward L in its triprotonated form are examined thoroughly. Crystallographic results show binding of fluoride in the C(3v)-symmetric cavity of [H(3)L](3+) where spherical anion fluoride is in tricoordinated geometry via (N-H)(+)···F interaction in the complex [H(3)L(F)]·[F](2)·2H(2)O, (3). In the case of complexes [H(3)L(OTs)]·[OTs](2), (4) and [H(3)L(OTs)]·[NO(3)]·[OTs], (5), tetrahedral p-toluenesulphonate ion is engulfed in the cavity of [H(3)L](3+) via (N-H)(+)···O interactions. Interestingly, complex [(H(3)L)(2)(SiF(6))]·[BF(4)](4)·CH(3)OH·H(2)O, (6) shows encapsulation of octahedral hexafluorosilicate in the dimeric capsular assembly of two [H(3)L](3+) units, via a number of (N-H)(+)···F interactions. The kinetic parameters of L upon binding with different anions are evaluated using a potentiometric study in solution state. The potentiometric titration experiments in a polar protic methanol/water (1:1 v/v) binary solvent system show high affinity of the receptor toward more basic fluoride and acetate anions, with a lesser affinity for other inorganic anions (e.g., chloride, bromide, nitrate, sulfate, dihydrogenphosphate, and p-toluenesulphonate). © 2011 American Chemical Society

The Dionaea muscipula ammonium channel DmAMT1 provides NH₄⁺ uptake associated with Venus flytrap's prey digestion.

PubMed

Scherzer, Sönke; Krol, Elzbieta; Kreuzer, Ines; Kruse, Jörg; Karl, Franziska; von Rüden, Martin; Escalante-Perez, Maria; Müller, Thomas; Rennenberg, Heinz; Al-Rasheid, Khaled A S; Neher, Erwin; Hedrich, Rainer

2013-09-09

Ammonium transporter (AMT/MEP/Rh) superfamily members mediate ammonium uptake and retrieval. This pivotal transport system is conserved among all living organisms. For plants, nitrogen represents a macronutrient available in the soil as ammonium, nitrate, and organic nitrogen compounds. Plants living on extremely nutrient-poor soils have developed a number of adaptation mechanisms, including a carnivorous lifestyle. This study addresses the molecular nature, function, and regulation of prey-derived ammonium uptake in the Venus flytrap, Dionaea muscipula, one of the fastest active carnivores. The Dionaea muscipula ammonium transporter DmAMT1 was localized in gland complexes where its expression was upregulated upon secretion. These clusters of cells decorating the inner trap surface are engaged in (1) secretion of an acidic digestive enzyme cocktail and (2) uptake of prey-derived nutrients. Voltage clamp of Xenopus oocytes expressing DmAMT1 and membrane potential recordings with DmAMT1-expressing Dionaea glands were used to monitor and compare electrophysiological properties of DmAMT1 in vitro and in planta. DmAMT1 exhibited the hallmark biophysical properties of a NH4(+)-selective channel. At depolarized membrane potentials (Vm = 0), the Km (3.2 ± 0.3 mM) indicated a low affinity of DmAMT1 for ammonium that increased systematically with negative going voltages. Upon hyperpolarization to, e.g., -200 mV, a Km of 0.14 ± 0.015 mM documents the voltage-dependent shift of DmAMT1 into a NH4(+) transport system of high affinity. We suggest that regulation of glandular DmAMT1 and membrane potential readjustments of the endocrine cells provide for effective adaptation to varying, prey-derived ammonium sources. Copyright © 2013 Elsevier Ltd. All rights reserved.

Dopamine receptor contribution to the action of PCP, LSD and ketamine psychotomimetics.

PubMed

Seeman, P; Ko, F; Tallerico, T

2005-09-01

Although phencyclidine and ketamine are used to model a hypoglutamate theory of schizophrenia, their selectivity for NMDA receptors has been questioned. To determine the affinities of phencyclidine, ketamine, dizocilpine and LSD for the functional high-affinity state of the dopamine D2 receptor, D2High, their dissociation constants (Ki) were obtained on [3H]domperidone binding to human cloned dopamine D2 receptors. Phencyclidine had a high affinity for D2High with a Ki of 2.7 nM, in contrast to its low affinity for the NMDA receptor, with a Ki of 313 nM, as labeled by [3H]dizocilpine on rat striatal tissue. Ketamine also had a high affinity for D2High with a Ki of 55 nM, an affinity higher than its 3100 nM Ki for the NMDA sites. Dizocilpine had a Ki of 0.3 nM at D2High, but a Kd of 1.8 nM at the NMDA receptor. LSD had a Ki of 2 nM at D2High. Because the psychotomimetics had higher potency at D2High than at the NMDA site, the psychotomimetic action of these drugs must have a major contribution from D2 agonism. Because these drugs have a combined action on both dopamine receptors and NMDA receptors, these drugs, when given in vivo, test a combined hyperdopamine and hypoglutamate theory of psychosis.

NO Metabolites Levels in Human Red Blood Cells are Affected by Palytoxin, an Inhibitor of Na(+)/K(+)-ATPase Pump.

PubMed

Carelli-Alinovi, Cristiana; Tellone, Ester; Russo, Anna Maria; Ficarra, Silvana; Pirolli, Davide; Galtieri, Antonio; Giardina, Bruno; Misiti, Francesco

2014-01-01

Palytoxin (PTX), a marine toxin, represents an increasing hazard for human health. Despite its high toxicity for biological systems, the mechanisms triggered by PTX, are not well understood. The high affinity of PTX for erythrocyte Na(+)/K(+)-ATPase pump is largely known, and it indicates PTX as a sensitive tool to characterize the signal transducer role for Na(+)/K(+)-ATPase pump. Previously, it has been reported that in red blood cells (RBC), probably via a signal transduction generated by the formation of a PTX-Na(+)/K(+)-ATPase complex, PTX alters band 3 functions and glucose metabolism. The present study addresses the question of which other signaling pathways are regulated by Na(+)/K(+)-ATPase in RBC. Here it has been evidenced that PTX following its interaction with Na(+)/K(+)-ATPase pump, alters RBC morphology and this event is correlated to decreases by 30% in nitrites and nitrates levels, known as markers of plasma membrane eNOS activity. Orthovanadate (OV), an antagonist of PTX binding to Na(+)/K(+)-ATPase pump, was able to reverse the effects elicited by PTX. Finally, current investigation firstly suggests that Na(+)/K(+)-ATPase pump, following its interaction with PTX, triggers a signal transduction involved in NO metabolism regulation.

Identification and properties of steroid-binding proteins in nesting Chelonia mydas plasma.

PubMed

Ikonomopoulou, M P; Bradley, A J; Whittier, J M; Ibrahim, K

2006-11-01

We report for the first time the presence of a sex steroid-binding protein in the plasma of green sea turtles Chelonia mydas, which provides an insight into reproductive status. A high affinity, low capacity sex hormone steroid-binding protein was identified in nesting C. mydas and its thermal profile was established. In nesting C. mydas testosterone and oestradiol bind at 4 degrees C with high affinity (K (a) = 1.49 +/- 0.09 x 10(9) M(-1); 0.17 +/- 0.02 x 10(7) M(-1)) and low binding capacity (B (max) = 3.24 +/- 0.84 x 10(-5) M; 0.33 +/- 0.06 x 10(-4) M). The binding affinity and capacity of testosterone at 23 and 36 degrees C, respectively were similar to those determined at 4 degrees C. However, oestradiol showed no binding activity at 36 degrees C. With competition studies we showed that oestradiol and oestrone do not compete for binding sites. Furthermore, in nesting C. mydas plasma no high-affinity binding was observed for adrenocortical steroids (cortisol and corticosterone) and progesterone. Our results indicate that in nesting C. mydas plasma temperature has a minimal effect on the high-affinity binding of testosterone to sex steroid-binding protein, however, the high affinity binding of oestradiol to sex steroid-binding protein is abolished at a hypothetically high (36 degrees C) sea/ambient/body temperature. This suggests that at high core body temperatures most of the oestradiol becomes biologically available to the tissues rather than remaining bound to a high-affinity carrier.

Effect of Nitrite and Nitrate Concentrations on the Performance of AFB-MFC Enriched with High-Strength Synthetic Wastewater.

PubMed

Huang, Jian-Sheng; Yang, Ping; Li, Chong-Ming; Guo, Yong; Lai, Bo; Wang, Ye; Feng, Li; Zhang, Yun

2015-01-01

In order to study the effect of nitrite and nitrate on the performance of microbial fuel cell, a system combining an anaerobic fluidized bed (AFB) and a microbial fuel cell (MFC) was employed for high-strength nitrogen-containing synthetic wastewater treatment. Before this study, the AFB-MFC had been used to treat high-strength organic wastewater for about one year in a continuous flow mode. The results showed that when the concentrations of nitrite nitrogen and nitrate nitrogen were increased from 1700 mg/L to 4045 mg/L and 545 mg/L to 1427 mg/L, respectively, the nitrite nitrogen and nitrate nitrogen removal efficiencies were both above 99%; the COD removal efficiency went up from 60.00% to 88.95%; the voltage was about 375 ± 15 mV while the power density was at 70 ± 5 mW/m(2). However, when the concentrations of nitrite nitrogen and nitrate nitrogen were above 4045 mg/L and 1427 mg/L, respectively, the removal of nitrite nitrogen, nitrate nitrogen, COD, voltage, and power density were decreased to be 86%, 88%, 77%, 180 mV, and 17 mW/m(2) when nitrite nitrogen and nitrate nitrogen were increased to 4265 mg/L and 1661 mg/L. In addition, the composition of biogas generated in the anode chamber was analyzed by a gas chromatograph. Nitrogen gas, methane, and carbon dioxide were obtained. The results indicated that denitrification happened in anode chamber.

Identifying riparian sinks for watershed nitrate using soil surveys.

PubMed

Rosenblatt, A E; Gold, A J; Stolt, M H; Groffman, P M; Kellogg, D Q

2001-01-01

The capacity of riparian zones to serve as critical control locations for watershed nitrogen flux varies with site characteristics. Without a means to stratify riparian zones into different levels of ground water nitrate removal capacity, this variability will confound spatially explicit source-sink models of watershed nitrate flux and limit efforts to target riparian restoration and management. We examined the capability of SSURGO (1:15 840 Soil Survey Geographic database) map classifications (slope class, geomorphology, and/or hydric soil designation) to identify riparian sites with high capacity for ground water nitrate removal. The study focused on 100 randomly selected riparian locations in a variety of forested and glaciated settings within Rhode Island. Geomorphic settings included till, outwash, and organic/alluvial deposits. We defined riparian zones with "high ground water nitrate removal capacity" as field sites possessing both >10 m of hydric soil width and an absence of ground water surface seeps. SSURGO classification based on a combination of geomorphology and hydric soil status created two functionally distinct sets of riparian sites. More than 75% of riparian sites classified by SSURGO as organic/alluviumhydric or as outwash-hydric had field attributes that suggest a high capacity for ground water nitrate removal. In contrast, >85% of all till sites and nonhydric outwash sites had field characteristics that minimize the capacity for ground water nitrate removal. Comparing the STATSGO and SSURGO databases for a 64000-ha watershed, STATSGO grossly under-represented critical riparian features. We conclude that the SSURGO database can provide modelers and managers with important insights into riparian zone nitrogen removal potential.

The competitive advantage of a dual-transporter system.

PubMed

Levy, Sagi; Kafri, Moshe; Carmi, Miri; Barkai, Naama

2011-12-09

Cells use transporters of different affinities to regulate nutrient influx. When nutrients are depleted, low-affinity transporters are replaced by high-affinity ones. High-affinity transporters are helpful when concentrations of nutrients are low, but the advantage of reducing their abundance when nutrients are abundant is less clear. When we eliminated such reduced production of the Saccharomyces cerevisiae high-affinity transporters for phosphate and zinc, the elapsed time from the initiation of the starvation program until the lack of nutrients limited growth was shortened, and recovery from starvation was delayed. The latter phenotype was rescued by constitutive activation of the starvation program. Dual-transporter systems appear to prolong preparation for starvation and to facilitate subsequent recovery, which may optimize sensing of nutrient depletion by integrating internal and external information about nutrient availability.

Contamination potential of nitrogen compounds in the heterogeneous aquifers of the Choushui River alluvial fan, Taiwan

NASA Astrophysics Data System (ADS)

Jang, Cheng-Shin; Liu, Chen-Wuing

2005-10-01

This study aimed to analyze the contamination potential associated with the reactive transport of nitrate-N and ammonium-N in the Choushui River alluvial fan, Taiwan and to evaluate a risk region in developing a groundwater protection policy in 2021. In this area, an aquifer redox sequence provided a good understanding of the spatial distributions of nitrate-N and ammonium-N and of aerobic and anaerobic environments. Equiprobable hydraulic conductivity ( K) fields reproduced by geostatistical methods characterized the spatial uncertainty of contaminant transport in the heterogeneous aquifer. Nitrogen contamination potential fronts for high and low threshold concentrations based on a 95% risk probability were used to assess different levels of risk. The simulated result reveals that the spatial uncertainty of highly heterogeneous K fields governs the contamination potential assessment of the nitrogen compounds along the regional flow directions. The contamination potential of nitrate-N is more uncertain than that for ammonium-N. The high nitrate-N concentrations (≧ 3 mg/L) are prevalent in the aerobic environment. The low concentration nitrate-N plumes (0.5-3 mg/L) gradually migrate to the mid-fan area and to a maximum distance of 15 km from the aerobic region. The nitrate-N plumes pose a potential human health risk in the aerobic and anaerobic environments. The ammonium-N plumes remain stably confined to the distal-fan and partial mid-fan areas.

A HIGH-LEVEL CALCULATION OF THE PROTON AFFINITY OF DIBORANE

EPA Science Inventory

The experimental proton affinity of diborane (B2H6) is based on an unstable species, B2H,+, 4 which has been observed only at low temperatures. The present work calculates the proton 5 affinity of diborane using the Gaussian-3 method and other high-level compound ab initio 6 met...

Detection of Sequence-Specific Tyrosine Nitration of Manganese SOD and SERCA in Cardiovascular Disease and Aging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xu, Shanqin; Ying, Jia; Jiang, Bingbing

2006-06-01

Nitration of protein tyrosine residues (nY) is a marker of oxidative stress and may alter the biological activity of the modified proteins. The aim of this study was to develop antibodies towards site-specific nY-modified proteins and to use histochemical and immunoblotting to demonstrate protein nitration in tissues. Affinity-purified polyclonal antibodies towards peptides with known nY sites in MnSOD nY-34 and of two adjacent nY in the sarcoplasmic endoplasmic reticulum calcium ATPase (SERCA2 di-nY-294,295) were developed. Kidneys from rats infused with angiotensin II with known MnSOD nY and aorta from atherosclerotic rabbits and aging rat skeletal and cardiac sarcoplasmic reticulum withmore » known SERCA di-nY were used for positive controls. Staining for MnSOD nY-34 was most intense in distal renal tubules and collecting ducts. Staining of atherosclerotic aorta for SERCA2 di-nY was most intense in atherosclerotic plaques. Aging rat skeletal muscle and atherosclerotic aorta and cardiac atrium from human diabetic patients also stained positively. Staining was decreased by sodium dithionite that chemically reduces nitrotyrosine to aminotyrosine, and the antigenic nY-peptide blocked staining for each respective nY site, but not for the other. As previously demonstrated, immunoblotting failed to detect these modified proteins in whole tissue lysates, but did when the proteins were concentrated. Immunohistochemical staining for specific nY-modified tyrosine residues offers the ability to assess the effects of oxidant stress associated with pathological conditions on individual proteins whose function may be affected in specific tissue sites.« less

Solvation of actinide salts in water using a polarizable continuum model.

PubMed

Kumar, Narendra; Seminario, Jorge M

2015-01-29

In order to determine how actinide atoms are dressed when solvated in water, density functional theory calculations have been carried out to study the equilibrium structure of uranium plutonium and thorium salts (UO2(2+), PuO2(2+), Pu(4+), and Th(4+)) both in vacuum as well as in solution represented by a conductor-like polarizable continuum model. This information is of paramount importance for the development of sensitive nanosensors. Both UO2(2+) and PuO2(2+) ions show coordination number of 4-5 with counterions replacing one or two water molecules from the first coordination shell. On the other hand, Pu(4+), has a coordination number of 8 both when completely solvated and also in the presence of chloride and nitrate ions with counterions replacing water molecules in the first shell. Nitrates were found to bind more strongly to Pu(IV) than chloride anions. In the case of the Th(IV) ion, the coordination number was found to be 9 or 10 in the presence of chlorides. Moreover, the Pu(IV) ion shows greater affinity for chlorides than the Th(IV) ion. Adding dispersion and ZPE corrections to the binding energy does not alter the trends in relative stability of several conformers because of error cancelations. All structures and energetics of these complexes are reported.

Effect of nitrate concentration on filamentous bulking under low level of dissolved oxygen in an airlift inner circular anoxic-aerobic incorporate reactor.

PubMed

Su, Yiming; Zhang, Yalei; Zhou, Xuefei; Jiang, Ming

2013-09-01

This laboratory research investigated a possible cause of filamentous bulking under low level of dissolved oxygen conditions (dissolved oxygen value in aerobic zone maintained between 0.6-0.8 mg O2/L) in an airlift inner-circular anoxic-aerobic reactor. During the operating period, it was observed that low nitrate concentrations affected sludge volume index significantly. Unlike the existing hypothesis, the batch tests indicated that filamentous bacteria (mainly Thiothrix sp.) could store nitrate temporarily under carbon restricted conditions. When nitrate concentration was below 4 mg/L, low levels of carbon substrates and dissolved oxygen in the aerobic zone stimulated the nitrate-storing capacity of filaments. When filamentous bacteria riched in nitrate reached the anoxic zone, where they were exposed to high levels of carbon but limited nitrate, they underwent denitrification. However, when nonfilamentous bacteria were exposed to similar conditions, denitrification was restrained due to their intrinsic nitrate limitation. Hence, in order to avoid filamentous bulking, the nitrate concentration in the return sludge (from aerobic zone to the anoxic zone) should be above 4 mg/L, or alternatively, the nitrate load in the anoxic zone should be kept at levels above 2.7 mg NO(3-)-N/g SS.

Detection of ovomucoid-specific low-affinity IgE in infants and its relationship to eczema.

PubMed

Kawamoto, Norio; Kamemura, Norio; Kido, Hiroshi; Fukao, Toshiyuki

2017-06-01

Allergen-specific low-affinity IgE was previously detected in cord blood by a highly sensitive densely carboxylated protein (DCP) chip, but not by ImmunoCAP. Here, we investigated the presence of low-affinity IgE during the early life of infants and observed its relationship with eczema. We conducted a birth cohort study, collecting sera at birth and 6 and 14 months of age (n = 110). We monitored the ovomucoid (OM)- and egg white (EW)-specific IgE (sIgE) by ImmunoCAP or DCP chip and analyzed the antigen affinity of sIgE by binding inhibition assays in the presence or absence of a mild chaotropic agent, diethyl amine (DEA). The low- and high-affinity OM-sIgEs and sensitization risk factors were analyzed by a multivariate logistic analysis. The OM-sIgE measured by DCP chip significantly correlated with that measured by ImmunoCAP, but some samples assessed as OM-sIgE positive by DCP chip were considered OM-sIgE negative by ImmunoCAP. Binding inhibition analysis after DEA treatment was performed for participants judged as OM-sIgE positive by DCP chip at 14 M. The group assessed as negative for OM- and EW-sIgE by ImmunoCAP at 6 and 14 months showed a larger binding inhibition curve shift after DEA treatment than did the group assessed as positive at these times, indicating the presence of low-affinity sIgE antibodies at 14 months. The logistic regression analysis found that persistent eczema from 6 to 14 months is a significant risk factor for developing high-affinity, but not low-affinity, sIgE. Human infant peripheral blood contains allergen-specific low-affinity sIgE. Persistent eczema is related to the development of high-affinity, but not low-affinity, IgE. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Modeling nitrate from land surface to wells' perforations under agricultural land: success, failure, and future scenarios in a Mediterranean case study

NASA Astrophysics Data System (ADS)

Levy, Yehuda; Shapira, Roi H.; Chefetz, Benny; Kurtzman, Daniel

2017-07-01

Contamination of groundwater resources by nitrate leaching under agricultural land is probably the most troublesome agriculture-related water contamination worldwide. Contaminated areas often show large spatial variability of nitrate concentration in wells. In this study, we tried to assess whether this spatial variability can be characterized on the basis of land use and standard agricultural practices. Deep soil sampling (10 m) was used to calibrate vertical flow and nitrogen-transport numerical models of the unsaturated zone under different agricultural land uses. Vegetable fields (potato and strawberry) and deciduous orchards (persimmon) in the Sharon area overlying the coastal aquifer of Israel were examined. Average nitrate-nitrogen fluxes below vegetable fields were 210-290 kg ha-1 yr-1 and under deciduous orchards were 110-140 kg ha-1 yr-1. The output water and nitrate-nitrogen fluxes of the unsaturated-zone models were used as input data for a three-dimensional flow and nitrate-transport model in the aquifer under an area of 13.3 km2 of agricultural land. The area was subdivided into four agricultural land uses: vegetables, deciduous orchards, citrus orchards, and non-cultivated. Fluxes of water and nitrate-nitrogen below citrus orchards were taken from a previous study in the area. The groundwater flow model was calibrated to well heads by changing the hydraulic conductivity. The nitrate-transport model, which was fed by the above-mentioned models of the unsaturated zone, succeeded in reconstructing the average nitrate concentration in the wells. However, this transport model failed in calculating the high concentrations in the most contaminated wells and the large spatial variability of nitrate concentrations in the aquifer. To reconstruct the spatial variability and enable predictions, nitrate fluxes from the unsaturated zone were multiplied by local multipliers. This action was rationalized by the fact that the high concentrations in some wells cannot be explained by regular agricultural activity and are probably due to malfunctions in the well area. Prediction of the nitrate concentration 40 years in the future with three nitrogen-fertilization scenarios showed that (i) under the business as usual fertilization scenario, the nitrate concentration (as NO3-) will increase on average by 19 mg L-1; (ii) under a scenario of 25 % reduction of nitrogen fertilization, the nitrate concentration in the aquifer will stabilize; (iii) with a 50 % reduction of nitrogen fertilization, the nitrate concentration will decrease on average by 18 mg L-1.

Of microbes and men: Determining sources of nitrate in a high alpine catchment in the Front Range of Colorado and science outreach on alpine hydrology

NASA Astrophysics Data System (ADS)

Hafich, Katya A.

High elevation ecosystems throughout the Colorado Front Range are undergoing changes in biogeochemical cycling due to an increase in nitrogen deposition in precipitation and a changing climate. While nitrate concentrations continue to rise in surface water of the Green Lakes Valley (GLV) by 0.27 umol L-1 per year, atmospheric deposition of inorganic nitrogen has recently curtailed due to drought, leaving a gap in our understanding of the source of the increased export of nitrate. Here, we employ a novel triple isotope method, using Delta 17O-NO3- for the first time in an alpine catchment to quantify the terrestrial and atmospheric contribution of nitrate to numerous water types in GLV. Results show that nitrate in surface waters, including talus, soil water and rock glacier melt, is more than 75% terrestrial, with the strongest atmospheric signals present during snowmelt. Results suggest that alpine catchment biogeochemistry in GLV has transitioned to a net nitrification system.

Endogenous levels of nitrites and nitrates in wide consumption foodstuffs: Results of five years of official controls and monitoring.

PubMed

Iammarino, Marco; Di Taranto, Aurelia; Cristino, Marianna

2013-10-15

The massive introduction of nitrogen fertilisers, necessary to maximise the global food production, has brought about an increase of the residual amounts of nitrites and nitrates in the products. Notoriously, these compounds may exercise toxic effects. In this work the results obtained from 5years of official controls and monitoring focused on tracing quantifiable amounts of nitrites and nitrates in 1785 samples of meat, dairy, fish products and leafy vegetables are reported. A widespread presence of nitrates at low concentrations in foodstuffs was verified. High concentrations of nitrates were registered in some leafy vegetables and mussels samples, while high nitrites concentrations were registered in some spinach samples. The results confirmed the necessity to develop most controls and suggest the introduction of new legal limits related to some combinations contaminant/matrix. Such new limits may fill legislative gaps that may cause wrong interpretations of the results obtained during official controls. Copyright © 2012 Elsevier Ltd. All rights reserved.

Forensic applications of nitrogen and oxygen isotopes in tracing nitrate sources in urban environments

USGS Publications Warehouse

Silva, S.R.; Ging, P.B.; Lee, R.W.; Ebbert, J.C.; Tesoriero, A.J.; Inkpen, E.L.

2002-01-01

Ground and surface waters in urban areas are susceptible to nitrate contamination from septic systems, leaking sewer lines, and fertilizer applications. Source identification is a primary step toward a successful remediation plan in affected areas. In this respect, nitrogen and oxygen isotope ratios of nitrate, in conjunction with hydrologic data and water chemistry, have proven valuable in urban studies from Austin, Texas, and Tacoma, Washington. In Austin, stream water was sampled during stremflow and baseflow conditions to assess surface and subsurface sources of nitrate, respectively. In Tacoma, well waters were sampled in adjacent sewered and un-sewered areas to determine if locally high nitrate concentrations were caused by septic systems in the un-sewered areas. In both studies, sewage was identified as a nitrate source and mixing between sewage and other sources of nitrate was apparent. In addition to source identification, combined nitrogen and oxygen isotopes were important in determining the significance of denitrification, which can complicate source assessment by reducing nitrate concentrations and increasing ??15N values. The two studies illustrate the value of nitrogen and oxygen isotopes of nitrate for forensic applications in urban areas. ?? Published by Elsevier Science Ltd. on behalf of AEHS.

Nitrate, Nitrite, and Ammonium Variability in Drinking Water Distribution Systems.

PubMed

Schullehner, Jörg; Stayner, Leslie; Hansen, Birgitte

2017-03-09

Accurate assessments of exposure to nitrate in drinking water is a crucial part of epidemiological studies investigating long-term adverse human health effects. However, since drinking water nitrate measurements are usually collected for regulatory purposes, assumptions on (1) the intra-distribution system variability and (2) short-term (seasonal) concentration variability have to be made. We assess concentration variability in the distribution system of nitrate, nitrite, and ammonium, and seasonal variability in all Danish public waterworks from 2007 to 2016. Nitrate concentrations at the exit of the waterworks are highly correlated with nitrate concentrations within the distribution net or at the consumers' taps, while nitrite and ammonium concentrations are generally lower within the net compared with the exit of the waterworks due to nitrification. However, nitrification of nitrite and ammonium in the distribution systems only results in a relatively small increase in nitrate concentrations. No seasonal variation for nitrate, nitrite, or ammonium was observed. We conclude that nitrate measurements taken at the exit of the waterworks are suitable to calculate exposures for all consumers connected to that waterworks and that sampling frequencies in the national monitoring programme are sufficient to describe temporal variations in longitudinal studies.

Nitrate, Nitrite, and Ammonium Variability in Drinking Water Distribution Systems

PubMed Central

Schullehner, Jörg; Stayner, Leslie; Hansen, Birgitte

2017-01-01

Accurate assessments of exposure to nitrate in drinking water is a crucial part of epidemiological studies investigating long-term adverse human health effects. However, since drinking water nitrate measurements are usually collected for regulatory purposes, assumptions on (1) the intra-distribution system variability and (2) short-term (seasonal) concentration variability have to be made. We assess concentration variability in the distribution system of nitrate, nitrite, and ammonium, and seasonal variability in all Danish public waterworks from 2007 to 2016. Nitrate concentrations at the exit of the waterworks are highly correlated with nitrate concentrations within the distribution net or at the consumers’ taps, while nitrite and ammonium concentrations are generally lower within the net compared with the exit of the waterworks due to nitrification. However, nitrification of nitrite and ammonium in the distribution systems only results in a relatively small increase in nitrate concentrations. No seasonal variation for nitrate, nitrite, or ammonium was observed. We conclude that nitrate measurements taken at the exit of the waterworks are suitable to calculate exposures for all consumers connected to that waterworks and that sampling frequencies in the national monitoring programme are sufficient to describe temporal variations in longitudinal studies. PMID:28282914

Hydrogen isotope fractionation in wood-producing avocado seedlings: Biological constraints to paleoclimatic interpretations of δD values in tree ring cellulose nitrate

NASA Astrophysics Data System (ADS)

Terwilliger, Valery J.; Deniro, Michael J.

1995-12-01

Climatic reconstructions from the δD values of wood cellulose nitrate have been compromised because it is unclear whether the isotopic ratios are affected only by temperature or by temperature and humidity. To quantify the effect of humidity on the δD values of leaf and wood cellulose nitrate, we grew avocados (Persea americana Mill. cv. Mexican) from seed at high and low humidities until they set wood. The source water for seed production was isotopically the same as the source water for seedling propagation. The δD values of leaf cellulose nitrate were related to those of leaf water, which were, in turn, influenced by humidity ( P < 0.01). The δD values of wood cellulose nitrate were unrelated to those of leaf water or any other indicator of humidity, but were related to the δD values of water in wood ( P ⩽ 0.05). The δD values of wood cellulose nitrate were identical in three out of five pairs of low and high humidity treatments. These results suggest humidity cannot be reliably inferred from δD values in wood cellulose nitrate. The δD values of cellulose nitrate in both leaves and wood appear to have been influenced by the incorporation of stored metabolites into cellulose. Trees, like avocado seedlings, have considerable post-photosynthetic organic reserves that can be tapped for growth. Conditions that stimulate use of post-photosynthetic carbon reserves are varied for trees. Significant contributions from these reserves could lead to erroneous temperature inferences from δD values of wood cellulose nitrate.

Nitrate radical oxidation of γ-terpinene: hydroxy nitrate, total organic nitrate, and secondary organic aerosol yields

NASA Astrophysics Data System (ADS)

Slade, Jonathan H.; de Perre, Chloé; Lee, Linda; Shepson, Paul B.

2017-07-01

Polyolefinic monoterpenes represent a potentially important but understudied source of organic nitrates (ONs) and secondary organic aerosol (SOA) following oxidation due to their high reactivity and propensity for multi-stage chemistry. Recent modeling work suggests that the oxidation of polyolefinic γ-terpinene can be the dominant source of nighttime ON in a mixed forest environment. However, the ON yields, aerosol partitioning behavior, and SOA yields from γ-terpinene oxidation by the nitrate radical (NO3), an important nighttime oxidant, have not been determined experimentally. In this work, we present a comprehensive experimental investigation of the total (gas + particle) ON, hydroxy nitrate, and SOA yields following γ-terpinene oxidation by NO3. Under dry conditions, the hydroxy nitrate yield = 4(+1/-3) %, total ON yield = 14(+3/-2) %, and SOA yield ≤ 10 % under atmospherically relevant particle mass loadings, similar to those for α-pinene + NO3. Using a chemical box model, we show that the measured concentrations of NO2 and γ-terpinene hydroxy nitrates can be reliably simulated from α-pinene + NO3 chemistry. This suggests that NO3 addition to either of the two internal double bonds of γ-terpinene primarily decomposes forming a relatively volatile keto-aldehyde, reconciling the small SOA yield observed here and for other internal olefinic terpenes. Based on aerosol partitioning analysis and identification of speciated particle-phase ON applying high-resolution liquid chromatography-mass spectrometry, we estimate that a significant fraction of the particle-phase ON has the hydroxy nitrate moiety. This work greatly contributes to our understanding of ON and SOA formation from polyolefin monoterpene oxidation, which could be important in the northern continental US and the Midwest, where polyolefinic monoterpene emissions are greatest.

Molybdenum Availability Is Key to Nitrate Removal in Contaminated Groundwater Environments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thorgersen, Michael P.; Lancaster, W. Andrew; Vaccaro, Brian J.

2015-05-15

The concentrations of molybdenum (Mo) and 25 other metals were measured in groundwater samples from 80 wells on the Oak Ridge Reservation (ORR) (Oak Ridge, TN), many of which are contaminated with nitrate, as well as uranium and various other metals. Moreover, the concentrations of nitrate and uranium were in the ranges of 0.1 μM to 230 mM and <0.2 nM to 580 μM, respectively. Most metals examined had significantly greater median concentrations in a subset of wells that were highly contaminated with uranium (≥126 nM). They included cadmium, manganese, and cobalt, which were 1,300- to 2,700-fold higher. A notablemore » exception, however, was Mo, which had a lower median concentration in the uranium-contaminated wells. This is significant, because Mo is essential in the dissimilatory nitrate reduction branch of the global nitrogen cycle. It is required at the catalytic site of nitrate reductase, the enzyme that reduces nitrate to nitrite. Furthermore, more than 85% of the groundwater samples contained less than 10 nM Mo, whereas concentrations of 10 to 100 nM Mo were required for efficient growth by nitrate reduction for twoPseudomonasstrains isolated from ORR wells and by a model denitrifier,Pseudomonas stutzeriRCH2. Higher concentrations of Mo tended to inhibit the growth of these strains due to the accumulation of toxic concentrations of nitrite, and this effect was exacerbated at high nitrate concentrations. The relevance of these results to a Mo-based nitrate removal strategy and the potential community-driving role that Mo plays in contaminated environments are discussed.« less

Assessing the impacts of future climate conditions on the effectiveness of winter cover crops in reducing nitrate loads into the Chesapeake Bay Watersheds using SWAT model

USDA-ARS?s Scientific Manuscript database

Winter cover crops (WCCs) have been widely implemented in the Coastal Plain of the Chesapeake Bay watershed (CBW) due to their high effectiveness at reducing nitrate loads. However, future climate conditions (FCCs) are expected to exacerbate water quality degradation in the CBW by increasing nitrat...

Development of a Small-Scale, High Efficiency Bioremediation System for Removing Nitrate from Nursery Runoff Water

USDA-ARS?s Scientific Manuscript database

Nitrate concentrations in runoff water from the nursery ranged from 70 to 253 mg NO3-N/L. An estimated 62 to 67% of the nitrate applied during fertigation events left the production site in runoff water. Irrigation losses during these events accounted for 36 to 49% of the amount applied, with flow r...

Vascular effects of dietary nitrate (as found in green leafy vegetables and beetroot) via the nitrate-nitrite-nitric oxide pathway.

PubMed

Lidder, Satnam; Webb, Andrew J

2013-03-01

The discovery that dietary (inorganic) nitrate has important vascular effects came from the relatively recent realization of the 'nitrate-nitrite-nitric oxide (NO) pathway'. Dietary nitrate has been demonstrated to have a range of beneficial vascular effects, including reducing blood pressure, inhibiting platelet aggregation, preserving or improving endothelial dysfunction, enhancing exercise performance in healthy individuals and patients with peripheral arterial disease. Pre-clinical studies with nitrate or nitrite also show the potential to protect against ischaemia-reperfusion injury and reduce arterial stiffness, inflammation and intimal thickness. However, there is a need for good evidence for hard endpoints beyond epidemiological studies. Whilst these suggest reduction in cardiovascular risk with diets high in nitrate-rich vegetables (such as a Mediterranean diet), others have suggested possible small positive and negative associations with dietary nitrate and cancer, but these remain unproven. Interactions with other nutrients, such as vitamin C, polyphenols and fatty acids may enhance or inhibit these effects. In order to provide simple guidance on nitrate intake from different vegetables, we have developed the Nitrate 'Veg-Table' with 'Nitrate Units' [each unit being 1 mmol of nitrate (62 mg)] to achieve a nitrate intake that is likely to be sufficient to derive benefit, but also to minimize the risk of potential side effects from excessive ingestion, given the current available evidence. The lack of data concerning the long term effects of dietary nitrate is a limitation, and this will need to be addressed in future trials. © 2012 The Authors. British Journal of Clinical Pharmacology © 2012 The British Pharmacological Society.

Characterization of two ammonia-oxidizing bacteria isolated from reactors operated with low dissolved oxygen concentrations.

PubMed

Park, H-D; Noguera, D R

2007-05-01

To obtain ammonia-oxidizing bacterial (AOB) strains inhabiting low dissolved oxygen (DO) environments and to characterize them to better understand their function and ecology. Using a serial dilution method, two AOB strains (ML1 and NL7) were isolated from chemostat reactors operated with low DO concentrations (0.12-0.24 mg l(-1)). Phylogenetically, strains ML1 and NL7 are affiliated to AOB within the Nitrosomonas europaea and Nitrosomonas oligotropha lineages, respectively. Kinetically, strain ML1 had high affinity for oxygen (0.24 +/- 0.13 mg l(-1)) and low affinity for ammonia (1.62 +/- 0.97 mg N l(-1)), while strain NL7 had high affinity for ammonia (0.48 +/- 0.35 mg l(-1)), but a surprisingly low affinity for oxygen (1.22 +/- 0.43 mg l(-1)). A co-culture experiment was used to iteratively estimate decay constants for both strains. The results indicated that AOB without high affinity for oxygen may have other mechanisms to persist in low DO environments, with high affinity for ammonia being important. This study provides a method to determine AOB growth kinetic parameters without assuming or neglecting decay constant. And, this is the first report on oxygen affinity constant of a N. oligotropha strain.

Heteropolyacids: An Efficient Catalyst for Synthesis of CL-20

NASA Astrophysics Data System (ADS)

Bayat, Yadollah; Mokhtari, Javad; Farhadian, Nafiseh; Bayat, Mohammad

2012-04-01

CL-20, a high-energy material with a cage-like structure, is considered the most powerful explosive today. It is usually prepared via nitration with concentrated nitric and sulfuric acid, but this technique pollutes the environment. In this article, CL-20 was synthesized by nitration of 2,6,8,12-tetraacetyl 2,4,6,8,10,12-hexaazatetracyclo[5,5,0,03,11,05,9]dodecane (TAIW) using a clean nitrating agent, heteropolyacids. Using the new nitrating agent caused the elimination of concentrated sulfuric acid during the reaction. This is an environmentally friendly technique.

Pharmacological and gene regulation properties point to the SlHAK5 K+ transporter as a system for high-affinity Cs+ uptake in tomato plants.

PubMed

Ródenas, Reyes; Nieves-Cordones, Manuel; Rivero, Rosa M; Martinez, Vicente; Rubio, Francisco

2018-04-01

Potassium (K + ) and cesium (Cs + ) are chemically similar but while K + is an essential nutrient, Cs + can be toxic for living organisms, plants included. Two different situations could lead to problems derived from the presence of Cs + in agricultural systems: (1) presence of Cs + at high concentrations that could produce toxic effects on plants, (2) presence of micromolar concentrations of radiocesium, which can be accumulated in the plant and affect animal and human health through the food chain. While K + uptake has been well described in tomato plants, information on molecular mechanisms involved in Cs + accumulation in this species is absent. Here, we show that in tomato plants, high concentrations of Cs + produce deficiency of K + but do not induce high-affinity K + uptake or the gene encoding the high-affinity K + transporter SlHAK5. At these concentrations, Cs + uptake takes place through a Ca 2+ -sensitive pathway, probably a non-selective cation channel. At micromolar concentrations, Cs + is accumulated by a high-affinity uptake system upregulated in K + -starved plants. This high-affinity Cs + uptake shares features with high-affinity K + uptake. It is sensitive to NH 4 + and insensitive to Ba 2+ and Ca 2+ and its presence parallels the pattern of SlHAK5 expression. Moreover, blockers of reactive oxygen species and ethylene action repress SlHAK5 and negatively regulate both high-affinity K + and Cs + uptake. Thus, we propose that SlHAK5 contributes to Cs + uptake from micromolar concentrations in tomato plants and can constitute a pathway for radiocesium transfer from contaminated areas to the food chain. © 2017 Scandinavian Plant Physiology Society.

Monthly variability and possible sources of nitrate in ground water beneath mixed agricultural land use, Suwannee and Lafayette Counties, Florida

USGS Publications Warehouse

Katz, Brian G.; Böhlke, J.K.

2000-01-01

In an area of mixed agricultural land use in Suwannee and Lafayette Counties of northern Florida, water samples were collected monthly from 14 wells tapping the Upper Floridan aquifer during July 1998 through June 1999 to assess hydrologic and land-use factors affecting the variability in nitrate concentrations in ground water. Unusually high amounts of rainfall in September and October 1998 (43.5 centimeters total for both months) resulted in an increase in water levels in all wells in October 1998. This was followed by unusually low amounts of rainfall during November 1998 through May 1999, when rainfall was 40.7 centimeters below 30-year mean monthly values. The presence of karst features (sinkholes, springs, solution conduits) and the highly permeable sands that overlie the Upper Floridan aquifer provide for rapid movement of water containing elevated nitrate concentrations to the aquifer. Nitrate was the dominant form of nitrogen in ground water collected at all sites and nitrate concentrations ranged from less than 0.02 to 22 milligrams per liter (mg/L), as nitrogen. Water samples from most wells showed substantial monthly or seasonal fluctuations in nitrate concentrations. Generally, water samples from wells with nitrate concentrations higher than 10 mg/L showed the greatest amount of monthly fluctuation. For example, water samples from six of eight wells had monthly nitrate concentrations that varied by at least 5 mg/L during the study period. Water from most wells with lower nitrate concentrations (less than 6 mg/L) also showed large monthly fluctuations. For instance, nitrate concentrations in water from four sites showed monthly variations of more than 50 percent. Large fluctuations in nitrate concentrations likely result from seasonal agricultural practices (fertilizer application and animal waste spreading) at a particular site. For example, an increase in nitrate concentrations observed in water samples from seven sites in February or March 1999 most likely results from application of synthetic fertilizers during the late winter months. Lower nitrate concentrations were detected in water samples from five of eight wells sampled during high-flow conditions for the Suwannee River in March 1998 compared to low-flow conditions in November 1998. Evidence for reduction of nitrate due to denitrification reactions was observed at one site (AC-1), as indicated by elevated concentrations of nitrogen gas and a corresponding increase in nitrogen isotope (d15N-NO3) values with a decrease in nitrate concentrations. Denitrification is unlikely at other sites based on the presence of dissolved oxygen concentrations greater than 2 mg/L in ground water and no observed trend between nitrate concentrations and values d15N-NO3 values. Nitrate was the dominant nitrogen species in most monthly rainfall samples; however, ammonium concentrations were similar or greater than nitrate during November and December 1998. During February through May 1999, both nitrate and ammonium concentrations were substantially higher in monthly rainfall samples collected at the study area compared to mean monthly concentrations at the Bradford Forest site located east of the study area, which is part of the National Atmospheric Deposition Program/National Trends Network. Also, higher nitrogen deposition rates in the study area compared to those at Bradford Forest could indicate that substantial amounts of ammonia are volatilized from fertilizers and animal wastes, released to the atmosphere, and incorporated as nitrate and ammonium in rainfall deposited in the middle Suwannee River Basin. Ground-water samples from most sites had d15N-NO3 values that indicated a mixture of inorganic and organic sources of nitrogen, which corresponded to multiple land uses where both synthetic fertilizers and manure are used on fields near these sites. Distinct d15N-NO3 signatures, however, were observed at some sites. For example, water samples from areas of row-crop farmin

Drinking-Water Nitrate, Methemoglobinemia, and Global Burden of Disease: A Discussion

PubMed Central

Fewtrell, Lorna

2004-01-01

On behalf of the World Health Organization (WHO), I have undertaken a series of literature-based investigations examining the global burden of disease related to a number of environmental risk factors associated with drinking water. In this article I outline the investigation of drinking-water nitrate concentration and methemoglobinemia. The exposure assessment was based on levels of nitrate in drinking water greater than the WHO guideline value of 50 mg/L. No exposure–response relationship, however, could be identified that related drinking-water nitrate level to methemoglobinemia. Indeed, although it has previously been accepted that consumption of drinking water high in nitrates causes methemoglobinemia in infants, it appears now that nitrate may be one of a number of co-factors that play a sometimes complex role in causing the disease. I conclude that, given the apparently low incidence of possible water-related methemoglobinemia, the complex nature of the role of nitrates, and that of individual behavior, it is currently inappropriate to attempt to link illness rates with drinking-water nitrate levels. PMID:15471727

Nitratreduktion in einem quartären Grundwasserleiter in Ostwestfalen, NRW

NASA Astrophysics Data System (ADS)

Wisotzky, Frank; Wohnlich, Stefan; Böddeker, Martin

2018-06-01

Groundwater chemistry and sediment chemistry are characterized in a catchment of a water works in the lower terrace of the river Ems, in east-Westphalia (Germany). In spite of strong nitrate input in the shallow groundwater, the nitrate concentration in the wells is very low, suggesting strong nitrate reduction. The aims of this study were to determine the type of nitrate reduction and to determine the depth of these reactions. As part of the groundwater investigation, soil samples were also used. All soil samples have low sulfide-sulfur contents but high organic-carbon contents. The shallow groundwater has nitrate concentrations up to 185 mg/l. Enhanced carbon-dioxide concentrations in the deeper part of the aquifer indicate a dominant organotrophic nitrate reduction. Data from a multi-level observation well show a 25% lithotrophic and 75% organotrophic nitrate reduction. Investigations of nitrate isotopes (δ15N-NO3-values and δ18O-NO3-values) and sulfate isotopes (δ34S-SO4-values and δ18O-SO4-values) support the findings.

Unexpectedly low nitrogen acquisition and absence of root architecture adaptation to nitrate supply in a Medicago truncatula highly branched root mutant

PubMed Central

Bourion, Virginie

2014-01-01

To complement N2 fixation through symbiosis, legumes can efficiently acquire soil mineral N through adapted root architecture. However, root architecture adaptation to mineral N availability has been little studied in legumes. Therefore, this study investigated the effect of nitrate availability on root architecture in Medicago truncatula and assessed the N-uptake potential of a new highly branched root mutant, TR185. The effects of varying nitrate supply on both root architecture and N uptake were characterized in the mutant and in the wild type. Surprisingly, the root architecture of the mutant was not modified by variation in nitrate supply. Moreover, despite its highly branched root architecture, TR185 had a permanently N-starved phenotype. A transcriptome analysis was performed to identify genes differentially expressed between the two genotypes. This analysis revealed differential responses related to the nitrate acquisition pathway and confirmed that N starvation occurred in TR185. Changes in amino acid content and expression of genes involved in the phenylpropanoid pathway were associated with differences in root architecture between the mutant and the wild type. PMID:24706718

Unexpectedly low nitrogen acquisition and absence of root architecture adaptation to nitrate supply in a Medicago truncatula highly branched root mutant.

PubMed

Bourion, Virginie; Martin, Chantal; de Larambergue, Henri; Jacquin, Françoise; Aubert, Grégoire; Martin-Magniette, Marie-Laure; Balzergue, Sandrine; Lescure, Geoffroy; Citerne, Sylvie; Lepetit, Marc; Munier-Jolain, Nathalie; Salon, Christophe; Duc, Gérard

2014-06-01

To complement N2 fixation through symbiosis, legumes can efficiently acquire soil mineral N through adapted root architecture. However, root architecture adaptation to mineral N availability has been little studied in legumes. Therefore, this study investigated the effect of nitrate availability on root architecture in Medicago truncatula and assessed the N-uptake potential of a new highly branched root mutant, TR185. The effects of varying nitrate supply on both root architecture and N uptake were characterized in the mutant and in the wild type. Surprisingly, the root architecture of the mutant was not modified by variation in nitrate supply. Moreover, despite its highly branched root architecture, TR185 had a permanently N-starved phenotype. A transcriptome analysis was performed to identify genes differentially expressed between the two genotypes. This analysis revealed differential responses related to the nitrate acquisition pathway and confirmed that N starvation occurred in TR185. Changes in amino acid content and expression of genes involved in the phenylpropanoid pathway were associated with differences in root architecture between the mutant and the wild type. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Stable silver/biopolymer hybrid plasmonic nanostructures for high performance surface enhanced raman scattering (SERS)

USDA-ARS?s Scientific Manuscript database

Silver/biopolymer nanoparticles were prepared by adding 100 mg silver nitrate to 2% polyvinyl alcohol solution and reduced the silver nitrate into silver ion using 2 % trisodium citrate for high performance Surface Enhanced Raman Scattering (SERS) substrates. Optical properties of nanoparticle were ...

Computational modeling and molecular imprinting for the development of acrylic polymers with high affinity for bile salts.

PubMed

Yañez, Fernando; Chianella, Iva; Piletsky, Sergey A; Concheiro, Angel; Alvarez-Lorenzo, Carmen

2010-02-05

This work has focused on the rational development of polymers capable of acting as traps of bile salts. Computational modeling was combined with molecular imprinting technology to obtain networks with high affinity for cholate salts in aqueous medium. The screening of a virtual library of 18 monomers, which are commonly used for imprinted networks, identified N-(3-aminopropyl)-methacrylate hydrochloride (APMA.HCl), N,N-diethylamino ethyl methacrylate (DEAEM) and ethyleneglycol methacrylate phosphate (EGMP) as suitable functional monomers with medium-to-high affinity for cholic acid. The polymers were prepared with a fix cholic acid:functional monomer mole ratio of 1:4, but with various cross-linking densities. Compared to polymers prepared without functional monomer, both imprinted and non-imprinted microparticles showed a high capability to remove sodium cholate from aqueous medium. High affinity APMA-based particles even resembled the performance of commercially available cholesterol-lowering granules. The imprinting effect was evident in most of the networks prepared, showing that computational modeling and molecular imprinting can act synergistically to improve the performance of certain polymers. Nevertheless, both the imprinted and non-imprinted networks prepared with the best monomer (APMA.HCl) identified by the modeling demonstrated such high affinity for the template that the imprinting effect was less important. The fitting of adsorption isotherms to the Freundlich model indicated that, in general, imprinting increases the population of high affinity binding sites, except when the affinity of the functional monomer for the target molecule is already very high. The cross-linking density was confirmed as a key parameter that determines the accessibility of the binding points to sodium cholate. Materials prepared with 9% mol APMA and 91% mol cross-linker showed enough affinity to achieve binding levels of up to 0.4 mmol g(-1) (i.e., 170 mg g(-1)) under flow (1 mL min(-1)) of 0.2 mM sodium cholate solution. Copyright 2009 Elsevier B.V. All rights reserved.

Hydrogeochemical investigation to understand nitrate movement in groundwater of volcanic island, Korea

NASA Astrophysics Data System (ADS)

Kwon, E. H.; Park, J.; Chung, E.; Kang, B. R.; Park, W. B.; Woo, N. C.

2017-12-01

Groundwater is the sole-source of water supply in the volcanic island, Jeju-do, Korea. Since early 1990s, the nitrate contamination of groundwater has increased especially in the western part of the island. High level of nitrate in water can cause not only health risk to human body but also environmental side effect such as eutrophication and algal bloom in the coastal area. Several studies have done to estimate nitrate contamination in groundwater of local areas, but none of them dealt with nitrate movement with flow paths. So, this study aimed to determine the source and migration of nitrate in groundwater in the Gosan area, located in the western part of Jeju island through seasonal monitoring of hydrogeochemistry and stable isotope analyses from pumping and monitoring wells. Water samples including rainfall and groundwater are measured for major ions (Ca, Na, K, Mg, SO4, HCO3, NO3, Cl, etc.) and stable isotopes (i.e., δ2H, δ18O, δ18O-NO3, δ15N-NO3). From the monitoring data, we could evaluate hydrochemical change during nitrate contamination, and also could identify that groundwater in Gosan area is recharged mainly by regional flow from the high-altitude region. In future study, we will conduct additional seasonal monitoring from the multi-depth monitoring wells and will use statistical analysis to understand pollution sources and paths specifically.

Occupation of low-affinity cholecystokinin (CCK) receptors by CCK activates signal transduction and stimulates amylase secretion in pancreatic acinar cells.

PubMed

Vinayek, R; Patto, R J; Menozzi, D; Gregory, J; Mrozinski, J E; Jensen, R T; Gardner, J D

1993-03-10

Based on the effects of monensin on binding of 125I-CCK-8 and its lack of effect on CCK-8-stimulated amylase secretion we previously proposed that pancreatic acinar cells possess three classes of CCK receptors: high-affinity receptors, low-affinity receptors and very low-affinity receptors [1]. In the present study we treated pancreatic acini with carbachol to induce a complete loss of high-affinity CCK receptors and then examined the action of CCK-8 on inositol trisphosphate IP3(1,4,5), cytosolic calcium and amylase secretion in an effort to confirm and extend our previous hypothesis. We found that first incubating pancreatic acini with 10 mM carbachol decreased binding of 125I-CCK-8 measured during a second incubation by causing a complete loss of high-affinity CCK receptors with no change in the low-affinity CCK receptors. Carbachol treatment of acini, however, did not alter the action of CCK-8 on IP3(1,4,5), cytosolic calcium or amylase secretion or the action of CCK-JMV-180 on amylase secretion or on the supramaximal inhibition of amylase secretion caused by CCK-8. The present findings support our previous hypothesis that pancreatic acinar cells possess three classes of CCK receptors and suggest that high-affinity CCK receptors do not mediate the action of CCK-8 on enzyme secretion, that low-affinity CCK receptors may mediate the action of CCK on cytosolic calcium that does not involve IP3(1,4,5) and produce the upstroke of the dose-response curve for CCK-8-stimulated amylase secretion and that very low-affinity CCK receptors mediate the actions of CCK on IP3(1,4,5) and cytosolic calcium and produce the downstroke of the dose-response curve for CCK-8-stimulated amylase secretion. Moreover, CCK-JMV-180 is a full agonist for stimulating amylase secretion by acting at low-affinity CCK receptors and is an antagonist at very low-affinity CCK receptors.

Determination of dominant sources of nitrate contamination in transboundary (Russian Federation/Ukraine) catchment with heterogeneous land use.

PubMed

Vystavna, Y; Diadin, D; Grynenko, V; Yakovlev, V; Vergeles, Y; Huneau, F; Rossi, P M; Hejzlar, J; Knöller, K

2017-09-18

Nitrate contamination of surface water and shallow groundwater was studied in transboundary (Russia/Ukraine) catchment with heterogeneous land use. Dominant sources of nitrate contamination were determined by applying a dual δ 15 N-NO 3 and δ 18 O-NO 3 isotope approach, multivariate statistics, and land use analysis. Nitrate concentration was highly variable from 0.25 to 22 mg L -1 in surface water and from 0.5 to 100 mg L -1 in groundwater. The applied method indicated that sewage to surface water and sewage and manure to groundwater were dominant sources of nitrate contamination. Nitrate/chloride molar ratio was added to support the dual isotope signature and indicated the contribution of fertilizers to the nitrate content in groundwater. Groundwater temperature was found to be an additional indicator of manure and sewerage leaks in the shallow aquifer which has limited protection and is vulnerable to groundwater pollution.

In situ nitrate measurements capture short-term variability and seasonal transitions during a drought - flood year in the Mississippi River Basin

NASA Astrophysics Data System (ADS)

Pellerin, B. A.; Bergamaschi, B. A.; Saraceno, J.; Downing, B. D.; Crawford, C.; Gilliom, R.; Frederick, P.

2013-12-01

Nitrogen flux from the Mississippi River to the Gulf of Mexico has received considerable attention because it fuels primary production on the continental shelf and can contribute to the summer hypoxia observed in the Gulf. Accurately quantifying the load of nitrogen - particularly as nitrate - to the Gulf is critical for both predicting the size of the oxygen-depleted dead zone and establishing targets for N load reduction from the basin. Fluxes have been historically calculated with load estimation models using 5-10 years of discrete nitrate data collected approximately 12-18 times per year. These traditional monthly to biweekly sampling intervals often fail to adequately capture hydrologic pulses ranging from early snowmelt periods to short-duration rainfall events in small streams, but the ability to adequately resolve patterns in water quality in large rivers has received much less attention. The recent commercial availability of in situ optical sensors for nitrate, together with new techniques for data collection and analysis, provides an opportunity to measure nitrate concentration on time scales in which environmental conditions actually change. Data have been collected and analyzed from a USGS optical nitrate sensor deployed in the Mississippi River at Baton Rouge, Louisiana, since November 2011. Our nitrate data, collected at three hour intervals, shows a strong relationship to depth- and width-integrated discrete nitrate concentrations measured on 20 dates (r2=0.99, slope=1) after correcting for a consistent, small positive bias (0.10 mg/L). The close relationship between the in situ data measured on edge of the channel and the depth- and width-integrated sample suggests that the fixed sensor measurements provide a robust proxy for cross-sectional averaged nitrate concentrations at Baton Rouge under a range of flow conditions. Nitrate concentrations ranged from a low of 0.19 mg/L as N on September 11, 2012 to a high of 3.09 mg/L as N on July 12, 2013. This covers nearly the entire range of nitrate concentrations measured at Baton Rouge (2005-2013) and 30 miles upriver at St. Francisville (1996-2013). Seasonality in nitrate concentrations and discharge was observed, but daily values of discharge and nitrate concentrations reveal a decoupling both between dry and wet years and within a given year. Results from our study also suggest an anomalously high flush of nitrate from the upper basin in the wet spring of 2013, with higher than expected daily nitrate loads based on the daily runoff. A comparison of calculated (e.g. sensor) versus modeled spring nitrate loads reveals differences of up to 30% during certain months, although the implications of those difference for predicting the size of the Gulf hypoxia are not yet known.

Identifying functional zones of denitrification in heterogeneous aquifer systems by numerical simulations - a case study

NASA Astrophysics Data System (ADS)

Jang, E.; Kalbacher, T.; He, W.; Shao, H.; Schueth, C.; Kolditz, O.

2014-12-01

Nitrate contamination in shallow groundwater is still one of the common problems in many countries. Because of its high solubility and anionic nature, nitrate can easily leach through soil and persist in groundwater for decades. High nitrate concentration has been suggested as a major cause of accelerated eutrophication, methemoglobinemia and gastric cancer. There are several factors influencing the fate of nitrate in groundwater system, which is e.g. distribution of N- sources to soil and groundwater, distribution and amount of reactive substances maintaining denitrification, rate of nitrate degradation and its kinetics, and geological characteristics of the aquifer. Nitrate transport and redox transformation processes are closely linked to complex and spatially distributed physical and chemical interaction, therefore it is difficult to predict and quantify in the field and laboratory experiment. Models can play a key role in elucidation of nitrate reduction pathway in groundwater system and in the design and evaluation of field tests to investigate in situ remediation technologies as well. The goal of the current study is to predict groundwater vulnerability to nitrate, to identify functional zones of denitrification in heterogeneous aquifer systems and to describe the uncertainty of the predictions due to scale effects. For this aim, we developed a kinetic model using multi-component mass transport code OpenGeoSys coupling with IPhreeqc module of the geochemical solver PHREEQC. The developed model included sequential aerobic and nitrate-based respiration, multi-Monod kinetics, multi-species biogeochemical reactions, and geological characteristics of the groundwater aquifer. Moreover water-rock interaction such as secondary mineral precipitation was also included in this model. In this presentation, we focused on the general modelling approach and present the simulation results of nitrate transport simulation in a hypothetical aquifer systems based on data from Hessian Ried, an important groundwater resource for the densely populated Rhine-Main region in Germany.

High-Affinity Recombinant Antibody Fragments (Fabs) Can Be Applied in Peptide Enrichment Immuno-MRM Assays

PubMed Central

2015-01-01

High-affinity antibodies binding to linear peptides in solution are a prerequisite for performing immuno-MRM, an emerging technology for protein quantitation with high precision and specificity using peptide immunoaffinity enrichment coupled to stable isotope dilution and targeted mass spectrometry. Recombinant antibodies can be generated from appropriate libraries in high-throughput in an automated laboratory and thus may offer advantages over conventional monoclonal antibodies. However, recombinant antibodies are typically obtained as fragments (Fab or scFv) expressed from E. coli, and it is not known whether these antibody formats are compatible with the established protocols and whether the affinities necessary for immunocapture of small linear peptides can be achieved with this technology. Hence, we performed a feasibility study to ask: (a) whether it is feasible to isolate high-affinity Fabs to small linear antigens and (b) whether it is feasible to incorporate antibody fragments into robust, quantitative immuno-MRM assays. We describe successful isolation of high-affinity Fab fragments against short (tryptic) peptides from a human combinatorial Fab library. We analytically characterize three immuno-MRM assays using recombinant Fabs, full-length IgGs constructed from these Fabs, or traditional monoclonals. We show that the antibody fragments show similar performance compared with traditional mouse- or rabbit-derived monoclonal antibodies. The data establish feasibility of isolating and incorporating high-affinity Fabs into peptide immuno-MRM assays. PMID:24568200

High-affinity recombinant antibody fragments (Fabs) can be applied in peptide enrichment immuno-MRM assays.

PubMed

Whiteaker, Jeffrey R; Zhao, Lei; Frisch, Christian; Ylera, Francisco; Harth, Stefan; Knappik, Achim; Paulovich, Amanda G

2014-04-04

High-affinity antibodies binding to linear peptides in solution are a prerequisite for performing immuno-MRM, an emerging technology for protein quantitation with high precision and specificity using peptide immunoaffinity enrichment coupled to stable isotope dilution and targeted mass spectrometry. Recombinant antibodies can be generated from appropriate libraries in high-throughput in an automated laboratory and thus may offer advantages over conventional monoclonal antibodies. However, recombinant antibodies are typically obtained as fragments (Fab or scFv) expressed from E. coli, and it is not known whether these antibody formats are compatible with the established protocols and whether the affinities necessary for immunocapture of small linear peptides can be achieved with this technology. Hence, we performed a feasibility study to ask: (a) whether it is feasible to isolate high-affinity Fabs to small linear antigens and (b) whether it is feasible to incorporate antibody fragments into robust, quantitative immuno-MRM assays. We describe successful isolation of high-affinity Fab fragments against short (tryptic) peptides from a human combinatorial Fab library. We analytically characterize three immuno-MRM assays using recombinant Fabs, full-length IgGs constructed from these Fabs, or traditional monoclonals. We show that the antibody fragments show similar performance compared with traditional mouse- or rabbit-derived monoclonal antibodies. The data establish feasibility of isolating and incorporating high-affinity Fabs into peptide immuno-MRM assays.

A Fluorescent Protein Scaffold for Presenting Structurally Constrained Peptides Provides an Effective Screening System to Identify High Affinity Target-Binding Peptides

PubMed Central

Kadonosono, Tetsuya; Yabe, Etsuri; Furuta, Tadaomi; Yamano, Akihiro; Tsubaki, Takuya; Sekine, Takuya; Kuchimaru, Takahiro; Sakurai, Minoru; Kizaka-Kondoh, Shinae

2014-01-01

Peptides that have high affinity for target molecules on the surface of cancer cells are crucial for the development of targeted cancer therapies. However, unstructured peptides often fail to bind their target molecules with high affinity. To efficiently identify high-affinity target-binding peptides, we have constructed a fluorescent protein scaffold, designated gFPS, in which structurally constrained peptides are integrated at residues K131–L137 of superfolder green fluorescent protein. Molecular dynamics simulation supported the suitability of this site for presentation of exogenous peptides with a constrained structure. gFPS can present 4 to 12 exogenous amino acids without a loss of fluorescence. When gFPSs presenting human epidermal growth factor receptor type 2 (HER2)-targeting peptides were added to the culture medium of HER2-expressing cells, we could easily identify the peptides with high HER2-affinity and -specificity based on gFPS fluorescence. In addition, gFPS could be expressed on the yeast cell surface and applied for a high-throughput screening. These results demonstrate that gFPS has the potential to serve as a powerful tool to improve screening of structurally constrained peptides that have a high target affinity, and suggest that it could expedite the one-step identification of clinically applicable cancer cell-binding peptides. PMID:25084350

Bohr effect and temperature sensitivity of hemoglobins from highland and lowland deer mice.

PubMed

Jensen, Birgitte; Storz, Jay F; Fago, Angela

2016-05-01

An important means of physiological adaptation to environmental hypoxia is an increased oxygen (O2) affinity of the hemoglobin (Hb) that can help secure high O2 saturation of arterial blood. However, the trade-off associated with a high Hb-O2 affinity is that it can compromise O2 unloading in the systemic capillaries. High-altitude deer mice (Peromyscus maniculatus) have evolved an increased Hb-O2 affinity relative to lowland conspecifics, but it is not known whether they have also evolved compensatory mechanisms to facilitate O2 unloading to respiring tissues. Here we investigate the effects of pH (Bohr effect) and temperature on the O2-affinity of high- and low-altitude deer mouse Hb variants, as these properties can potentially facilitate O2 unloading to metabolizing tissues. Our experiments revealed that Bohr factors for the high- and low-altitude Hb variants are very similar in spite of the differences in O2-affinity. The Bohr factors of deer mouse Hbs are also comparable to those of other mammalian Hbs. In contrast, the high- and low-altitude variants of deer mouse Hb exhibited similarly low temperature sensitivities that were independent of red blood cell anionic cofactors, suggesting an appreciable endothermic allosteric transition upon oxygenation. In conclusion, high-altitude deer mice have evolved an adaptive increase in Hb-O2 affinity, but this is not associated with compensatory changes in sensitivity to changes in pH or temperature. Instead, it appears that the elevated Hb-O2 affinity in high-altitude deer mice is compensated by an associated increase in the tissue diffusion capacity of O2 (via increased muscle capillarization), which promotes O2 unloading. Copyright © 2016 Elsevier Inc. All rights reserved.

Utilization of urea and expression profiles of related genes in the dinoflagellate Prorocentrum donghaiense

PubMed Central

Jing, Xiaoli; Lin, Senjie; Zhang, Huan; Koerting, Claudia; Yu, Zhigang

2017-01-01

Urea has been shown to contribute more than half of total nitrogen (N) required by phytoplankton in some estuaries and coastal waters and to provide a substantial portion of the N demand for many harmful algal blooms (HABs) of dinoflagellates. In this study, we investigated the physiological and transcriptional responses in Prorocentrum donghaiense to changes in nitrate and urea availability. We found that this species could efficiently utilize urea as sole N source and achieve comparable growth rate and photosynthesis capability as it did under nitrate. These physiological parameters were markedly lower in cultures grown under nitrate- or urea-limited conditions. P. donghaiense N content was similarly low under nitrate- or urea-limited culture condition, but was markedly higher under urea-replete condition than under nitrate-replete condition. Carbon (C) content was consistently elevated under N-limited condition. Consequently, the C:N ratio was as high as 21:1 under nitrate- or urea-limitation, but 7:1 under urea-replete condition and 9:1 to 10:1 under nitrate-replete condition. Using quantitative reverse transcription PCR, we investigated the expression pattern for four genes involved in N transport and assimilation. The results indicated that genes encoding nitrate transport, urea hydrolysis, and nickel transporter gene were sensitive to changes in general N nutrient availability whereas the urea transporter gene responded much more strongly to changes in urea concentration. Taken together, our study shows the high bioavailability of urea, its impact on C:N stoichiometry, and the sensitivity of urea transporter gene expression to urea availability. PMID:29117255

Large scale reactive transport of nitrate across the surface water divide

NASA Astrophysics Data System (ADS)

Kortunov, E.; Lu, C.; Amos, R.; Grathwohl, P.

2016-12-01

Groundwater pollution caused by agricultural and atmospheric inputs is a pressing issue in environmental management worldwide. Various researchers have studied different aspects of nitrate contamination since the substantial increase of the agriculture pollution in the second half of the 20th century. This study addresses large scale reactive solute transport in a typical Germany hilly landscapes in a transect crossing 2 valleys: River Neckar and Ammer. The numerical model was constructed compromising a 2-D cross-section accounting for typical fractured mudstones and unconsolidated sediments. Flow modelling showed that the groundwater divide significantly deviates from the surface water divide providing conditions for inter-valley flow and transport. Reactive transport modelling of redox-sensitive solutes (e.g. agriculture nitrate and natural sulfate, DOC, ammonium) with MIN3P was used to elucidate source of nitrate in aquifers and rivers. Since both floodplains, in the Ammer and Neckar valley contain Holocene sediments relatively high in organic carbon, agricultural nitrate is reduced therein and does not reach the groundwater. However, nitrate applied in the hillslopes underlain by fractured oxidized mudrock is transported to the high yield sand and gravel aquifer in the Neckar valley. Therefore, the model predicts that nitrate in the Neckar valley comes, to a large extent, from the neighboring Ammer valley. Moreover, nitrate observed in the rivers and drains in the Ammer valley is very likely geogenic since frequent peat layers there release ammonium which is oxidized as it enters the surface water. Such findings are relevant for land and water quality management.

Utilization of urea and expression profiles of related genes in the dinoflagellate Prorocentrum donghaiense.

PubMed

Jing, Xiaoli; Lin, Senjie; Zhang, Huan; Koerting, Claudia; Yu, Zhigang

2017-01-01

Urea has been shown to contribute more than half of total nitrogen (N) required by phytoplankton in some estuaries and coastal waters and to provide a substantial portion of the N demand for many harmful algal blooms (HABs) of dinoflagellates. In this study, we investigated the physiological and transcriptional responses in Prorocentrum donghaiense to changes in nitrate and urea availability. We found that this species could efficiently utilize urea as sole N source and achieve comparable growth rate and photosynthesis capability as it did under nitrate. These physiological parameters were markedly lower in cultures grown under nitrate- or urea-limited conditions. P. donghaiense N content was similarly low under nitrate- or urea-limited culture condition, but was markedly higher under urea-replete condition than under nitrate-replete condition. Carbon (C) content was consistently elevated under N-limited condition. Consequently, the C:N ratio was as high as 21:1 under nitrate- or urea-limitation, but 7:1 under urea-replete condition and 9:1 to 10:1 under nitrate-replete condition. Using quantitative reverse transcription PCR, we investigated the expression pattern for four genes involved in N transport and assimilation. The results indicated that genes encoding nitrate transport, urea hydrolysis, and nickel transporter gene were sensitive to changes in general N nutrient availability whereas the urea transporter gene responded much more strongly to changes in urea concentration. Taken together, our study shows the high bioavailability of urea, its impact on C:N stoichiometry, and the sensitivity of urea transporter gene expression to urea availability.

Effects of melatonin on seedling growth, mineral nutrition, and nitrogen metabolism in cucumber under nitrate stress.

PubMed

Zhang, Ruimin; Sun, Yunkuo; Liu, Zeyu; Jin, Wen; Sun, Yan

2017-05-01

In China, excessive use of nitrogen fertilizers in glasshouses leads to nitrate accumulations in soil and plants, which then limits productivity. Melatonin, an evolutionarily highly conserved molecule, has a wide range of functions in plants. We analyzed the effects of melatonin pretreatment on the growth, mineral nutrition, and nitrogen metabolism in cucumber (Cucumis sativus L. "Jin You No. 1") when seedlings were exposed to nitrate stress. An application of 0.1 mmol/L melatonin significantly improved the growth of plants and reduced their susceptibility to damage due to high nitrate levels (0.6 mol/L) during the ensuing period of stress treatment. Although excess nitrate led to an increase in the concentrations of nitrogen, potassium, and calcium, as well as a decrease in levels of phosphorus and magnesium, exogenous melatonin generally had the opposite effect except for a further rise in calcium concentrations. Pretreatment also significantly reduced the accumulations of nitrate nitrogen and ammonium nitrogen and enhanced the activities of enzymes involved in nitrogen metabolism. Expression of Cs-NR and Cs-GOGAT, two genes that function in that metabolism, was greatly down-regulated when plants were exposed to 0.6 mol/L nitrate, but was up-regulated in plants that had received the 0.1 mmol/L melatonin pretreatment. Our results are the first evidence that melatonin has an important role in modulating the composition of mineral elements and nitrogen metabolism, thereby alleviating the inhibitory effect on growth normally associated with nitrate stress. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

New Synthesis and Tritium Labeling of a Selective Ligand for Studying High-affinity γ-Hydroxybutyrate (GHB) Binding Sites

PubMed Central

Vogensen, Stine B.; Marek, Aleš; Bay, Tina; Wellendorph, Petrine; Kehler, Jan; Bundgaard, Christoffer; Frølund, Bente; Pedersen, Martin H.F.; Clausen, Rasmus P.

2013-01-01

3-Hydroxycyclopent-1-enecarboxylic acid (HOCPCA, 1) is a potent ligand for the high-affinity GHB binding sites in the CNS. An improved synthesis of 1 together with a very efficient synthesis of [3H]-1 is described. The radiosynthesis employs in situ generated lithium trimethoxyborotritide. Screening of 1 against different CNS targets establishes a high selectivity and we demonstrate in vivo brain penetration. In vitro characterization of [3H]-1 binding shows high specificity to the high-affinity GHB binding sites. PMID:24053696

Direct Measurement of Equilibrium Constants for High-Affinity Hemoglobins

PubMed Central

Kundu, Suman; Premer, Scott A.; Hoy, Julie A.; Trent, James T.; Hargrove, Mark S.

2003-01-01

The biological functions of heme proteins are linked to their rate and affinity constants for ligand binding. Kinetic experiments are commonly used to measure equilibrium constants for traditional hemoglobins comprised of pentacoordinate ligand binding sites and simple bimolecular reaction schemes. However, kinetic methods do not always yield reliable equilibrium constants with more complex hemoglobins for which reaction mechanisms are not clearly understood. Furthermore, even where reaction mechanisms are clearly understood, it is very difficult to directly measure equilibrium constants for oxygen and carbon monoxide binding to high-affinity (KD ≪ 1 μM) hemoglobins. This work presents a method for direct measurement of equilibrium constants for high-affinity hemoglobins that utilizes a competition for ligands between the "target" protein and an array of "scavenger" hemoglobins with known affinities. This method is described for oxygen and carbon monoxide binding to two hexacoordinate hemoglobins: rice nonsymbiotic hemoglobin and Synechocystis hemoglobin. Our results demonstrate that although these proteins have different mechanisms for ligand binding, their affinities for oxygen and carbon monoxide are similar. Their large affinity constants for oxygen, 285 and ∼100 μM−1 respectively, indicate that they are not capable of facilitating oxygen transport. PMID:12770899

Direct Measurement of T Cell Receptor Affinity and Sequence from Naïve Anti-Viral T Cells

PubMed Central

Zhang, Shuqi; Parker, Patricia; Ma, Keyue; He, Chenfeng; Shi, Qian; Cui, Zhonghao; Williams, Chad; Wendel, Ben S.; Meriwether, Amanda; Salazar, Mary A.; Jiang, Ning

2016-01-01

T cells recognize and kill a myriad of pathogen-infected or cancer cells using a diverse set of T cell receptors (TCR). The affinity of TCR to cognate antigen is of high interest in adoptive T cell transfer immunotherapy and antigen-specific T cell repertoire immune profiling because it is widely known to correlate with downstream T cell responses. Here, we introduce the in situ TCR affinity and sequence test (iTAST) for simultaneous measurement of TCR affinity and sequence from single primary CD8+ T cells in human blood. We demonstrate that the repertoire of primary antigen-specific T cells from pathogen inexperienced individuals has a surprisingly broad affinity range of 1000-fold composed of diverse TCR sequences. Within this range, samples from older individuals contained a reduced frequency of high affinity T cells compared to young individuals, demonstrating an age-related effect of T cell attrition that could cause holes in the repertoire. iTAST should enable the rapid selection of high affinity TCRs ex vivo for adoptive immunotherapy and measurement of T cell response for immune monitoring applications. PMID:27252176

Impact of ammonium nitrate and sodium nitrate on tadpoles of Alytes obstetricans.

PubMed

Garriga, Núria; Montori, A; Llorente, G A

2017-07-01

The presence of pesticides, herbicides and fertilisers negatively affect aquatic communities in general, and particularly amphibians in their larval phase, even though sensitivity to pollutants is highly variable among species. The Llobregat Delta (Barcelona, Spain) has experienced a decline of amphibian populations, possibly related to the reduction in water quality due to the high levels of farming activity, but also to habitat loss and alteration. We studied the effects of increasing ammonium nitrate and sodium nitrate levels on the survival and growth rate of Alytes obstetricans tadpoles under experimental conditions. We exposed larvae to increasing concentrations of nitrate and ammonium for 14 days and then exposed them to water without pollutants for a further 14 days. Only the higher concentrations of ammonium (>33.75 mg/L) caused larval mortality. The growth rate of larvae was reduced at ≥22.5 mg/L NH 4 + , although individuals recovered and even increased their growth rate once exposure to the pollutant ended. The effect of nitrate on growth rate was detected at ≥80 mg/L concentrations, and the growth rate reduction in tadpoles was even observed during the post-exposure phase. The concentrations of ammonium with adverse effects on larvae are within the range levels found in the study area, while the nitrate concentrations with some adverse effect are close to the upper range limit of current concentrations in the study area. Therefore, only the presence of ammonium in the study area is likely to be considered of concern for the population of this species, even though the presence of nitrate could cause some sublethal effects. These negative effects could have an impact on population dynamics, which in this species is highly sensitive to larval mortality due to its small clutch size and prolonged larval period compared to other anuran amphibians.

DNA sequence determinants controlling affinity, stability and shape of DNA complexes bound by the nucleoid protein Fis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hancock, Stephen P.; Stella, Stefano; Cascio, Duilio

The abundant Fis nucleoid protein selectively binds poorly related DNA sequences with high affinities to regulate diverse DNA reactions. Fis binds DNA primarily through DNA backbone contacts and selects target sites by reading conformational properties of DNA sequences, most prominently intrinsic minor groove widths. High-affinity binding requires Fis-stabilized DNA conformational changes that vary depending on DNA sequence. In order to better understand the molecular basis for high affinity site recognition, we analyzed the effects of DNA sequence within and flanking the core Fis binding site on binding affinity and DNA structure. X-ray crystal structures of Fis-DNA complexes containing variable sequencesmore » in the noncontacted center of the binding site or variations within the major groove interfaces show that the DNA can adapt to the Fis dimer surface asymmetrically. We show that the presence and position of pyrimidine-purine base steps within the major groove interfaces affect both local DNA bending and minor groove compression to modulate affinities and lifetimes of Fis-DNA complexes. Sequences flanking the core binding site also modulate complex affinities, lifetimes, and the degree of local and global Fis-induced DNA bending. In particular, a G immediately upstream of the 15 bp core sequence inhibits binding and bending, and A-tracts within the flanking base pairs increase both complex lifetimes and global DNA curvatures. Taken together, our observations support a revised DNA motif specifying high-affinity Fis binding and highlight the range of conformations that Fis-bound DNA can adopt. Lastly, the affinities and DNA conformations of individual Fis-DNA complexes are likely to be tailored to their context-specific biological functions.« less

DNA sequence determinants controlling affinity, stability and shape of DNA complexes bound by the nucleoid protein Fis

DOE PAGES

Hancock, Stephen P.; Stella, Stefano; Cascio, Duilio; ...

2016-03-09

The abundant Fis nucleoid protein selectively binds poorly related DNA sequences with high affinities to regulate diverse DNA reactions. Fis binds DNA primarily through DNA backbone contacts and selects target sites by reading conformational properties of DNA sequences, most prominently intrinsic minor groove widths. High-affinity binding requires Fis-stabilized DNA conformational changes that vary depending on DNA sequence. In order to better understand the molecular basis for high affinity site recognition, we analyzed the effects of DNA sequence within and flanking the core Fis binding site on binding affinity and DNA structure. X-ray crystal structures of Fis-DNA complexes containing variable sequencesmore » in the noncontacted center of the binding site or variations within the major groove interfaces show that the DNA can adapt to the Fis dimer surface asymmetrically. We show that the presence and position of pyrimidine-purine base steps within the major groove interfaces affect both local DNA bending and minor groove compression to modulate affinities and lifetimes of Fis-DNA complexes. Sequences flanking the core binding site also modulate complex affinities, lifetimes, and the degree of local and global Fis-induced DNA bending. In particular, a G immediately upstream of the 15 bp core sequence inhibits binding and bending, and A-tracts within the flanking base pairs increase both complex lifetimes and global DNA curvatures. Taken together, our observations support a revised DNA motif specifying high-affinity Fis binding and highlight the range of conformations that Fis-bound DNA can adopt. Lastly, the affinities and DNA conformations of individual Fis-DNA complexes are likely to be tailored to their context-specific biological functions.« less

Organic Nitrates: A Complex Family of Atmospheric Trace Constituents

NASA Astrophysics Data System (ADS)

Ballschmiter, K.; Fischer, R. G.; Grünert, A.; Kastler, J.; Schneider, M.; Woidich, S.

2003-04-01

Biogenic and geogenic hydrocarbons are the precursors of organic nitrates that are formed as tropospheric photo-oxidation products in the presence of NOx. Air chemistry leads to a very complex pattern of nitric acid esters: alkyl nitrates, aryl-alkyl nitrates, and bifunctional nitrates like alkyl dinitrates, hydroxy alkyl nitrates and carbonyl alkyl nitrates. We have analyzed the pattern of organic nitrates in air samples after adsorption/thermal desorption (low volume sampling-LVS) or adsorption/solvent desorption (high volume sampling-HVS) by capillary gas chromatography with electron capture (ECD) and mass spectrometric detection (MSD) using air aliquotes of 100 up to 3000 liters on column. The complexity of the organic nitrates found in air requires a group pre-separation by normal phase liquid chromatography. A detection limit per compound of 0.005 ppt(v) is achieved by our approach. We have synthesized a broad spectrum of organic nitrates as reference compounds. Air samples were taken from central Europe, the US West (Utah, Nevada, California), and the North- and South Atlantic including Antarctica. Levels and patterns of the regional and global occurrence of the various groups of C1-C12 organic nitrates including dinitrates and hydroxy nitrates and nitrates of isoprene (2-methylbutadiene) are presented. Werner G., J. Kastler, R. Looser, K. Ballschmiter: "Organic nitrates of isoprene as atmospheric trace compounds" Angewandte Chemie - International Edition (1999) 38: 1634-1637. Woidich S., O. Froescheis, O. Luxenhofer, K. Ballschmiter: "EI- and NCI-mass spectrometry of arylalkyl nitrates and their occurrence in urban air" Fresenius J. Anal. Chem. (1999) 364 : 91-99. Kastler, J; Jarman, W; Ballschmiter, K.: "Multifunctional organic nitrates as constituents in European and US urban photo-smog" Fresenius J. Anal. Chem. (2000) 368:244-249. Schneider M., K. Ballschmiter: "C3-C14 alkyl nitrates in remote South Atlantic air" Chemosphere (1999) 38: 233-244. Fischer, R G; Kastler, J; Ballschmiter, K.: "Levels and pattern of alkyl nitrates, multifunctional alkyl nitrates, and halocarbons in the air over the Atlantic Ocean" J. Geophys. Research (2000) 105:14473-14494. Fischer R.G. , R. Weller, H.-W. Jacobi, K. Ballschmiter: "Levels and patterns of volatile organic nitrates and halocarbons in the air at Neumayer Station (70°), Antarctic" Chemosphere (2002) 48:981-992

Oxygen Isotopic Composition of Nitrate and Sulfate in Fog and River water in Podocarpus National Forest, Ecuador

NASA Astrophysics Data System (ADS)

Brothers, L. A.; Fabian, P.; Thiemens, M. H.

2006-12-01

The eastern slopes of the Andean rainforests of Ecuador possess some of the highest plant biodiversity found on the planet; however, these ecosystems are in jeopardy because region is experiences one of the highest deforestation rates in South America. This rainforest characterized by high acidity and low nutrient soils and experiences natural process which are both destabilizing and stabilizing to biodiversity rendering this a unique, though sensitive environment. There is increased concern that anthropogenic activities are affecting rainforests and could lead to higher extinction rates, changes in the biodiversity and far reaching effects on the global troposphere. Measurements of nitrate and sulfate in rain and fog water have shown periods of elevated concentrations in the Podocarpus National Park near Loja, Ecuador. These high episodes contribute to annual deposition rates that are comparable to polluted central Europe. Significant anthropogenic sources near this region are lacking and it is believed that the majority of the nitrate and sulfate pollution can be attributed to biomass burning in the Amazon basin. Concentration measurements do not elucidate the source of high nitrate and sulfate pollution; however, by measuring all three stable isotopes of oxygen in nitrate and sulfate from fog and river water provides a new way to examine the impacts of biomass burning on the region. By using stable isotope techniques atmospheric nitrate and sulfate can be resolved from terrestrial sources. This provides an unique way to trace the contributions from the biomass burning and farming sources. Current research at the field station monitors sulfate and nitrate concentrations in rain and fog water by standard methods to investigate water and nutrient pathways along with data from satellite and ground based remote sensing, in-situ observations and numerical models.

Concentration of Nitrate near the Surface of Frozen Salt Solutions

NASA Astrophysics Data System (ADS)

Michelsen, R. R. H.; Marrocco, H. A.

2017-12-01

The photolysis of nitrate near the surface of snow and ice in Earth's environment results in the emission of nitrogen oxides (NO, NO2 and, in acidic snow, HONO) and OH radicals. As a result, nitrate photolysis affects the composition and oxidative capacity of the overlying atmosphere. Photolysis yields depend in part on how much nitrate is close enough to the surface to be photolyzed. These concentrations are assumed to be higher than the concentrations of nitrate that are measured in melted snow and ice samples. However, near-surface concentrations of nitrate have not been directly measured. In this work, laboratory studies of the concentration of nitrate in frozen aqueous solutions are described. Individual aqueous solutions of nitric acid, sodium nitrate, and magnesium nitrate were mixed. Attenuated total reflection infrared spectroscopy was utilized to measure the nitrate and liquid water signals within 200 - 400 nm of the lower surface of frozen samples. Temperature was varied from -18°C to -2°C. In addition to the amount of nitrate observed, changes to the frozen samples' morphology with annealing are discussed. Nitrate concentrations near the lower surface of these frozen solutions are high: close to 1 M at warmer temperatures and almost 4 M at the coldest temperature. Known freezing point depression data describe the observed concentrations better than ideal solution thermodynamics, which overestimate concentration significantly at colder temperatures. The implications for modeling the chemistry of snow are discussed. Extending and relating this work to the interaction of gas-phase nitric acid with the surfaces of vapor-deposited ice will also be explored.

Hydrological controls on DOC  :  nitrate resource stoichiometry in a lowland, agricultural catchment, southern UK

NASA Astrophysics Data System (ADS)

Heppell, Catherine M.; Binley, Andrew; Trimmer, Mark; Darch, Tegan; Jones, Ashley; Malone, Ed; Collins, Adrian L.; Johnes, Penny J.; Freer, Jim E.; Lloyd, Charlotte E. M.

2017-09-01

The role that hydrology plays in governing the interactions between dissolved organic carbon (DOC) and nitrogen in rivers draining lowland, agricultural landscapes is currently poorly understood. In light of the potential changes to the production and delivery of DOC and nitrate to rivers arising from climate change and land use management, there is a pressing need to improve our understanding of hydrological controls on DOC and nitrate dynamics in such catchments. We measured DOC and nitrate concentrations in river water of six reaches of the lowland river Hampshire Avon (Wiltshire, southern UK) in order to quantify the relationship between BFI (BFI) and DOC : nitrate molar ratios across contrasting geologies (Chalk, Greensand, and clay). We found a significant positive relationship between nitrate and BFI (p < 0. 0001), and a significant negative relationship between DOC and BFI (p < 0. 0001), resulting in a non-linear negative correlation between DOC : nitrate molar ratio and BFI. In the Hampshire Avon, headwater reaches which are underlain by clay and characterized by a more flashy hydrological regime are associated with DOC : nitrate ratios > 5 throughout the year, whilst groundwater-dominated reaches underlain by Chalk, with a high BFI have DOC : nitrate ratios in surface waters that are an order of magnitude lower (< 0.5). Our analysis also reveals significant seasonal variations in DOC : nitrate transport and highlights critical periods of nitrate export (e.g. winter in sub-catchments underlain by Chalk and Greensand, and autumn in drained, clay sub-catchments) when DOC : nitrate molar ratios are low, suggesting low potential for in-stream uptake of inorganic forms of nitrogen. Consequently, our study emphasizes the tight relationship between DOC and nitrate availability in agricultural catchments, and further reveals that this relationship is controlled to a great extent by the hydrological setting.

Reactions Between Water Soluble Organic Acids and Nitrates in Atmospheric Aerosols: Recycling of Nitric Acid and Formation of Organic Salts

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Bingbing; Laskin, Alexander

Atmospheric particles often include a complex mixture of nitrate and secondary organic materials accumulated within the same individual particles. Nitrate as an important inorganic component can be chemically formed in the atmosphere. For instance, formation of sodium nitrate (NaNO3) and calcium nitrate Ca(NO3)2 when nitrogen oxide and nitric acid (HNO3) species react with sea salt and calcite, respectively. Organic acids contribute a significant fraction of photochemically formed secondary organics that can condense on the preexisting nitrate-containing particles. Here, we present a systematic microanalysis study on chemical composition of laboratory generated particles composed of water soluble organic acids and nitrates (i.e.more » NaNO3 and Ca(NO3)2) investigated using computer controlled scanning electron microscopy with energy dispersive analysis of X-rays (CCSEM/EDX) and Fourier transform infrared micro-spectroscopy (micro-FTIR). The results show that water-soluble organic acids can react with nitrates releasing gaseous HNO3 during dehydration process. These reactions are attributed to acid displacement of nitrate with weak organic acids driven by the evaporation of HNO3 into gas phase due to its relatively high volatility. The reactions result in significant nitrate depletion and formation of organic salts in mixed organic acids/nitrate particles that in turn may affect their physical and chemical properties relevant to atmospheric environment and climate. Airborne nitrate concentrations are estimated by thermodynamic calculations corresponding to various nitrate depletions in selected organic acids of atmospheric relevance. The results indicate a potential mechanism of HNO3 recycling, which may further affect concentrations of gas- and aerosol-phase species in the atmosphere and the heterogeneous reaction chemistry between them.« less

Photocatalytic reduction of nitrate using titanium dioxide for regeneration of ion exchange brine

PubMed Central

Yang, Ting; Doudrick, Kyle; Westerhoff, Paul

2016-01-01

Nitrate is often removed from groundwater by ion exchange (IX) before its use as drinking water. Accumulation of nitrate in IX brine reduces the efficiency of IX regeneration and the useful life of the regeneration brine. For the first time, we present a strategy to photocatalytically reduce nitrate in IX brine, thereby extending the use of the brine. Titanium dioxide (Evonik P90), acting as photocatalyst, reduced nitrate effectively in both synthetic brines and sulfate-removed IX brine when formic acid (FA) was used as the hole scavenger (i.e., electron donor) and the initial FA to nitrate molar ratio (IFNR) was 5.6. Increasing the NaCl level in the synthetic brine slowed the nitrate reduction rate without affecting byproduct selectivity of ammonium and gaseous N species (e.g., N2, N2O). In a non-modified IX brine, nitrate removal was greatly inhibited owing to the presence of sulfate, which competed with nitrate for active surface sites on P90 and induced aggregation of P90 nanoparticles. After removing sulfate through barium sulfate precipitation, nitrate was effectively reduced; approximately 3.6 × 1024 photons were required to reduce each mole of nitrate to 83% N Gases and 17% NH4+. To make optimum use of FA and control the residual FA level in treated brine, the IFNR was varied. High IFNRs (e.g., 4, 5.6) were found to be more efficient for nitrate reduction but left higher residual FA in brine. IX column tests were performed to investigate the impact of residual FA for brine reuse. The residual FA in the brine did not significantly affect the nitrate removal capacity of IX resins, and formate contamination of treated water could be eliminated by rinsing with one bed volume of fresh brine. PMID:23276425

Transformation of Nitrate and Toluene in Groundwater by Sulfur Modified Iron(SMI-III)

NASA Astrophysics Data System (ADS)

Lee, W.; Park, S.; Lim, J.; Hong, U.; Kwon, S.; Kim, Y.

2009-12-01

In Korea, nitrate and benzene, toluene, ethylbenzene, and xylene isomers (BTEX) are frequently detected together as ground water contaminants. Therefore, a system simultaneously treating both nitrate (inorganic compound) and BTEX (organic compounds) is required to utilize groundwater as a water resource. In this study, we investigated the efficiency of Sulfur Modified Iron (SMI-III) in treating both nitrate and BTEX contaminated groundwater. Based on XRD (X-Ray Diffraction) analysis, the SMI-III is mainly composed of Fe3O4, S, and Fe. A series of column tests were conducted at three different empty bed contact times (EBCTs) for each compound. During the experiments, removal efficiency for both nitrate and toluene were linearly correlated with EBCT, suggesting that SMI-III have an ability to transform both nitrate and toluene. The concentration of SO42- and oxidation/reduction potential (ORP) were also measured. After exposed to nitrate contaminated groundwater, the composition of SMI-III was changed to Fe2O3, Fe3O4, Fe, and Fe0.95S1.05. The trends of effluent sulfate concentrations were inversely correlated with effluent nitrate concentrations, while the trends of ORP values, having the minimum values of -480 mV, were highly correlated with effluent nitrate concentrations. XRD analysis before and after exposed to nitrate contaminated groundwater, sulfate production, and nitrite detection as a reductive transformation by-product of nitrate suggest that nitrate is reductively transformed by SMI-III. Interestingly, in the toluene experiments, the trends of ORP values were inversely correlated with effluent toluene concentrations, suggesting that probably degrade through oxidation reaction. Consequently, nitrate and toluene probably degrade through reduction and oxidation reaction, respectively and SMI-III could serve as both electron donor and acceptor.

Multi-scale nitrate transport in a sandstone aquifer system under intensive agriculture

NASA Astrophysics Data System (ADS)

Paradis, Daniel; Ballard, Jean-Marc; Lefebvre, René; Savard, Martine M.

2018-03-01

Nitrate transport in heterogeneous bedrock aquifers is influenced by mechanisms that operate at different spatial and temporal scales. To understand these mechanisms in a fractured sandstone aquifer with high porosity, a groundwater-flow and nitrate transport model—reproducing multiple hydraulic and chemical targets—was developed to explain the actual nitrate contamination observed in groundwater and surface water in a study area on Prince Edward Island, Canada. Simulations show that nitrate is leached to the aquifer year-round, with 61% coming from untransformed and transformed organic sources originating from fertilizers and manure. This nitrate reaches the more permeable shallow aquifer through fractures in weathered sandstone that represent only 1% of the total porosity (17%). Some of the nitrate reaches the underlying aquifer, which is less active in terms of groundwater flow, but most of it is drained to the main river. The river-water quality is controlled by the nitrate input from the shallow aquifer. Groundwater in the underlying aquifer, which has long residence times, is also largely influenced by the diffusion of nitrate in the porous sandstone matrix. Consequently, following a change of fertilizer application practices, water quality in domestic wells and the river would change rapidly due to the level of nitrate found in fractures, but a lag time of up to 20 years would be necessary to reach a steady level due to diffusion. This demonstrates the importance of understanding nitrate transport mechanisms when designing effective agricultural and water management plans to improve water quality.

Chronic nitrate exposure alters reproductive physiology in fathead minnows.

PubMed

Kellock, Kristen A; Moore, Adrian P; Bringolf, Robert B

2018-01-01

Nitrate is a ubiquitous aquatic pollutant that is commonly associated with eutrophication and dead zones in estuaries around the world. At high concentrations nitrate is toxic to aquatic life but at environmental concentrations it has also been purported as an endocrine disruptor in fish. To investigate the potential for nitrate to cause endocrine disruption in fish, we conducted a lifecycle study with fathead minnows (Pimephales promelas) exposed to nitrate (0, 11.3, and 56.5 mg/L (total nitrate-nitrogen (NO 3 -N)) from <24 h post hatch to sexual maturity (209 days). Body mass, condition factor, gonadal somatic index (GSI), incidence of intersex, and vitellogenin induction were determined in mature male and female fish and plasma 11-keto testosterone (11-KT) was measured in males only. In nitrate-exposed males both 11-KT and vitellogenin were significantly induced when compared with controls. No significant differences occurred for body mass, condition factor, or GSI among males and intersex was not observed in any of the nitrate treatments. Nitrate-exposed females also had significant increases in vitellogenin compared to controls but no significant differences for mass, condition factor, or GSI were observed in nitrate exposed groups. Estradiol was used as a positive control for vitellogenin induction. Our findings suggest that environmentally relevant nitrate levels may disrupt steroid hormone synthesis and/or metabolism in male and female fish and may have implications for fish reproduction, watershed management, and regulation of nutrient pollution. Copyright © 2017 Elsevier Ltd. All rights reserved.

Nitrate intake and the risk of thyroid cancer and thyroid disease.

PubMed

Ward, Mary H; Kilfoy, Briseis A; Weyer, Peter J; Anderson, Kristin E; Folsom, Aaron R; Cerhan, James R

2010-05-01

Nitrate is a contaminant of drinking water in agricultural areas and is found at high levels in some vegetables. Nitrate competes with uptake of iodide by the thyroid, thus potentially affecting thyroid function. We investigated the association of nitrate intake from public water supplies and diet with the risk of thyroid cancer and self-reported hypothyroidism and hyperthyroidism in a cohort of 21,977 older women in Iowa who were enrolled in 1986 and who had used the same water supply for >10 years. We estimated nitrate ingestion from drinking water using a public database of nitrate measurements (1955-1988). Dietary nitrate intake was estimated using a food frequency questionnaire and levels from the published literature. Cancer incidence was determined through 2004. We found an increased risk of thyroid cancer with higher average nitrate levels in public water supplies and with longer consumption of water exceeding 5 mg/L nitrate-N (for >or=5 years at >5 mg/L, relative risk [RR] = 2.6 [95% confidence interval (CI) = 1.1-6.2]). We observed no association with prevalence of hypothyroidism or hyperthyroidism. Increasing intake of dietary nitrate was associated with an increased risk of thyroid cancer (highest vs. lowest quartile, RR = 2.9 [1.0-8.1]; P for trend = 0.046) and with the prevalence of hypothyroidism (odds ratio = 1.2 [95% CI = 1.1-1.4]), but not hyperthyroidism. Nitrate may play a role in the etiology of thyroid cancer and warrants further study.

Alleviation of proton toxicity by nitrate uptake specifically depends on nitrate transporter 1.1 in Arabidopsis.

PubMed

Fang, Xian Zhi; Tian, Wen Hao; Liu, Xing Xing; Lin, Xian Yong; Jin, Chong Wei; Zheng, Shao Jian

2016-07-01

Protons in acid soil are highly rhizotoxic to plants, but the mechanism of tolerance of plants to protons is largely unknown. Nitrate uptake by root cells is accompanied by the uptake of protons. Therefore, nitrate uptake transporters (NRTs) may be involved in plant tolerance to proton toxicity. We investigated the root nitrate uptake response to proton stress in Arabidopsis and its association with proton tolerance using NRT-related mutants and pharmacological methods. Lack of NRT1.1 in knockout nrt1.1 mutants led to impaired proton tolerance in nitrate-sufficient growth medium, whereas no difference was seen between wild-type plants and NRT1.2-, NRT2.1-, NRT2.2-, and NRT2.4-null mutants. Another nrt1.1 point mutant, which is defective in nitrate uptake but has a normal nitrate-sensing function, also had impaired proton tolerance compared with the wild-type plant. Furthermore, proton stress induced NRT1.1-mediated nitrate uptake. These results indicate that NRT1.1-conferred proton tolerance depends on nitrate uptake activity. In addition, the rooting medium was alkalified by wild-type plants, but not by knockout nrt1.1 mutants, and in pH-buffered medium, there were no differences in proton tolerance between wild-type plants and knockout nrt1.1 mutants. We conclude that NRT1.1-mediated nitrate uptake plays a crucial role in plant proton tolerance by alkalifying the rhizosphere. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

Preparation of well-adhered γ-Al 2O 3 washcoat on metallic wire mesh monoliths by electrophoretic deposition

NASA Astrophysics Data System (ADS)

Sun, Hong; Quan, Xie; Chen, Shuo; Zhao, Huimin; Zhao, Yazhi

2007-01-01

Washcoat deposited on metallic wire mesh monoliths was prepared using γ-alumina powders by electrophoretic deposition under a relatively low electric voltage. The microstructure, phase structure and adhesion of washcoat were investigated by SEM, XRD, ultrasonic vibration and thermal shock. The results showed that the loading and adhesion of washcoat were affected obviously by the properties of suspension, such as the zeta potential and the amount of adding binders. A small quantity of aluminum isopropoxide could promote the cohesive affinity of washcoat in thermal shock. The adhesion of washcoat in ultrasonic vibration could be reinforced by increasing calcined temperature and adding a certain aluminum particles. It was also found that the washcoat immersed metal nitrate has excellent vibration-resistant ability.

Control of Microbial Sulfide Production with Biocides and Nitrate in Oil Reservoir Simulating Bioreactors

PubMed Central

Xue, Yuan; Voordouw, Gerrit

2015-01-01

Oil reservoir souring by the microbial reduction of sulfate to sulfide is unwanted, because it enhances corrosion of metal infrastructure used for oil production and processing. Reservoir souring can be prevented or remediated by the injection of nitrate or biocides, although injection of biocides into reservoirs is not commonly done. Whether combined application of these agents may give synergistic reservoir souring control is unknown. In order to address this we have used up-flow sand-packed bioreactors injected with 2 mM sulfate and volatile fatty acids (VFA, 3 mM each of acetate, propionate and butyrate) at a flow rate of 3 or 6 pore volumes (PV) per day. Pulsed injection of the biocides glutaraldehyde (Glut), benzalkonium chloride (BAC) and cocodiamine was used to control souring. Souring control was determined as the recovery time (RT) needed to re-establish an aqueous sulfide concentration of 0.8–1 mM (of the 1.7–2 mM before the pulse). Pulses were either for a long time (120 h) at low concentration (long-low) or for a short time (1 h) at high concentration (short-high). The short-high strategy gave better souring control with Glut, whereas the long-low strategy was better with cocodiamine. Continuous injection of 2 mM nitrate alone was not effective, because 3 mM VFA can fully reduce both 2 mM nitrate to nitrite and N2 and, subsequently, 2 mM sulfate to sulfide. No synergy was observed for short-high pulsed biocides and continuously injected nitrate. However, use of continuous nitrate and long-low pulsed biocide gave synergistic souring control with BAC and Glut, as indicated by increased RTs in the presence, as compared to the absence of nitrate. Increased production of nitrite, which increases the effectiveness of souring control by biocides, is the most likely cause for this synergy. PMID:26696994

Increasing dietary nitrate has no effect on cancellous bone loss or fecal microbiome in ovariectomized rats.

PubMed

Conley, Melissa N; Roberts, Cooper; Sharpton, Thomas J; Iwaniec, Urszula T; Hord, Norman G

2017-05-01

Studies suggest diets rich in fruit and vegetables reduce bone loss, although the specific compounds responsible are unknown. Substrates for endogenous nitric oxide (NO) production, including organic nitrates and dietary nitrate, may support NO production in age-related conditions, including osteoporosis. We investigated the capability of dietary nitrate to improve NO bioavailability, reduce bone turnover and loss. Six-month-old Sprague Dawley rats [30 ovariectomized (OVX) and 10 sham-operated (sham)] were randomized into three groups: (i) vehicle (water) control, (ii) low-dose nitrate (LDN, 0.1 mmol nitrate/kg bw/day), or (iii) high-dose nitrate (HDN, 1.0 mmol nitrate/kg bw/day) for three weeks. The sham received vehicle. Serum bone turnover markers; bone mass, mineral density, and quality; histomorphometric parameters; and fecal microbiome were examined. Three weeks of LDN or HDN improved NO bioavailability in a dose-dependent manner. OVX resulted in cancellous bone loss, increased bone turnover, and fecal microbiome changes. OVX increased relative abundances of Firmicutes and decreased Bacteroideceae and Alcaligenaceae. Nitrate did not affect the skeleton or fecal microbiome. These data indicate that OVX affects the fecal microbiome and that the gut microbiome is associated with bone mass. Three weeks of nitrate supplementation does not slow bone loss or alter the fecal microbiome in OVX. © 2017 The Authors. Molecular Nutrition & Food Research published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Enterosalivary nitrate metabolism and the microbiome: Intersection of microbial metabolism, nitric oxide and diet in cardiac and pulmonary vascular health.

PubMed

Koch, Carl D; Gladwin, Mark T; Freeman, Bruce A; Lundberg, Jon O; Weitzberg, Eddie; Morris, Alison

2017-04-01

Recent insights into the bioactivation and signaling actions of inorganic, dietary nitrate and nitrite now suggest a critical role for the microbiome in the development of cardiac and pulmonary vascular diseases. Once thought to be the inert, end-products of endothelial-derived nitric oxide (NO) heme-oxidation, nitrate and nitrite are now considered major sources of exogenous NO that exhibit enhanced vasoactive signaling activity under conditions of hypoxia and stress. The bioavailability of nitrate and nitrite depend on the enzymatic reduction of nitrate to nitrite by a unique set of bacterial nitrate reductase enzymes possessed by specific bacterial populations in the mammalian mouth and gut. The pathogenesis of pulmonary hypertension (PH), obesity, hypertension and CVD are linked to defects in NO signaling, suggesting a role for commensal oral bacteria to shape the development of PH through the formation of nitrite, NO and other bioactive nitrogen oxides. Oral supplementation with inorganic nitrate or nitrate-containing foods exert pleiotropic, beneficial vascular effects in the setting of inflammation, endothelial dysfunction, ischemia-reperfusion injury and in pre-clinical models of PH, while traditional high-nitrate dietary patterns are associated with beneficial outcomes in hypertension, obesity and CVD. These observations highlight the potential of the microbiome in the development of novel nitrate- and nitrite-based therapeutics for PH, CVD and their risk factors. Copyright © 2017 Elsevier Inc. All rights reserved.

An artificial self-sufficient cytochrome P450 directly nitrates fluorinated tryptophan analogs with a different regio-selectivity.

PubMed

Zuo, Ran; Zhang, Yi; Huguet-Tapia, Jose C; Mehta, Mishal; Dedic, Evelina; Bruner, Steven D; Loria, Rosemary; Ding, Yousong

2016-05-01

Aromatic nitration is an immensely important industrial process to produce chemicals for a variety of applications, but it often suffers from multiple unsolved challenges. Enzymes as biocatalysts have been increasingly used for organic chemistry synthesis due to their high selectivity and environmental friendliness, but nitration has benefited minimally from the development of biocatalysis. In this work, we aimed to develop TxtE as practical biocatalysts for aromatic nitration. TxtE is a unique class I cytochrome P450 enzyme that nitrates the indole of l-tryptophan. To develop cost-efficient nitration processes, we fused TxtE with the reductase domains of CYP102A1 (P450BM3) and of P450RhF to create class III self-sufficient biocatalysts. The best engineered fusion protein was comparable with wild type TxtE in terms of nitration performance and other key biochemical properties. To demonstrate the application potential of the fusion enzyme, we nitrated 4-F-dl-tryptophan and 5-F-l-tryptophan in large scale enzymatic reactions. Tandem MS/MS and NMR analyses of isolated products revealed altered nitration sites. To our knowledge, these studies represent the first practice in developing biological nitration approaches and lay a solid basis to the use of TxtE-based biocatalysts for the production of valuable nitroaromatics. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Probability of nitrate contamination of recently recharged groundwaters in the conterminous United States

USGS Publications Warehouse

Nolan, B.T.; Hitt, K.J.; Ruddy, B.C.

2002-01-01

A new logistic regression (LR) model was used to predict the probability of nitrate contamination exceeding 4 mg/L in predominantly shallow, recently recharged ground waters of the United States. The new model contains variables representing (1) N fertilizer loading (p 2 = 0.875), indicating that the LR model fits the data well. The likelihood of nitrate contamination is greater in areas with high N loading and well-drained surficial soils over unconsolidated sand and gravels. The LR model correctly predicted the status of nitrate contamination in 75% of wells in a validation data set. Considering all wells used in both calibration and validation, observed median nitrate concentration increased from 0.24 to 8.30 mg/L as the mapped probability of nitrate exceeding 4 mg/L increased from less than or equal to 0.17 to > 0.83.

Existence of three subtypes of bradykinin B2 receptors in guinea pig.

PubMed

Seguin, L; Widdowson, P S; Giesen-Crouse, E

1992-12-01

We describe the binding of [3H]bradykinin to homogenates of guinea pig brain, lung, and ileum. Analysis of [3H]bradykinin binding kinetics in guinea pig brain, lung, and ileum suggests the existence of two binding sites in each tissue. The finding of two binding sites for [3H]bradykinin in ileum, lung, and brain was further supported by Scatchard analysis of equilibrium binding in each tissue. [3H]Bradykinin binds to a high-affinity site in brain, lung, and ileum (KD = 70-200 pM), which constitutes approximately 20% of the bradykinin binding, and to a second, lower-affinity site (0.63-0.95 nM), which constitutes the remaining 80% of binding. Displacement studies with various bradykinin analogues led us to subdivide the high- and lower-affinity sites in each tissue and to suggest the existence of three subtypes of B2 receptors in the guinea pig, which we classify as B2a, B2b, and B2c. Binding of [3H]bradykinin is largely to a B2b receptor subtype, which constitutes the majority of binding in brain, lung, and ileum and represents the lower-affinity site in our binding studies. Receptor subtype B2c constitutes approximately 20% of binding sites in the brain and lung and is equivalent to the high-affinity site in brain and lung. We suggest that a third subtype of B2 receptor (high-affinity site in ileum), B2a, is found only in the ileum. All three subtypes of B2 receptors display a high affinity for bradykinin, whereas they show different affinities for various bradykinin analogues displaying agonist or antagonist activities.(ABSTRACT TRUNCATED AT 250 WORDS)

Affinity-tuned ErbB2 or EGFR chimeric antigen receptor T cells exhibit an increased therapeutic index against tumors in mice

PubMed Central

Liu, Xiaojun; Jiang, Shuguang; Fang, Chongyun; Yang, Shiyu; Olalere, Devvora; Pequignot, Edward C.; Cogdill, Alexandria P.; Li, Na; Ramones, Melissa; Granda, Brian; Zhou, Li; Loew, Andreas; Young, Regina M.; June, Carl H.; Zhao, Yangbing

2015-01-01

Target-mediated toxicity is a major limitation in the development of chimeric antigen T cell receptors (CAR) for adoptive cell therapy of solid tumors. In this study, we developed a strategy to adjust the affinities of the scFv component of CAR to discriminate tumors overexpressing the target from normal tissues which express it at physiologic levels. A CAR-expressing T cell panel was generated with target antigen affinities varying over three orders of magnitude. High-affinity cells recognized target expressed at any level, including at levels in normal cells that were undetectable by flow cytometry. Affinity-tuned cells exhibited robust antitumor efficacy similar to high-affinity cells, but spared normal cells expressing physiologic target levels. The use of affinity-tuned scFvs offers a strategy to empower wider use of CAR T cells against validated targets widely overexpressed on solid tumors, including those considered undruggable by this approach. PMID:26330166

[Progresses in screening active compounds from herbal medicine by affinity chromatography].

PubMed

Feng, Ying-shu; Tong, Shan-shan; Xu, Xi-ming; Yu, Jiang-nan

2015-03-01

Affinity chromatography is a chromatographic method for separating molecules using the binding characteristics of the stationary phase with potential drug molecules. This method can be performed as a high throughput screening method and a chromatographic separation method to screen a variety of active drugs. This paper summarizes the history of affinity chromatography, screening technology of affinity chromatography, and application of affinity chromatography in screening bio-active compounds in herbal medicines, and then discusses its application prospects, in order to broaden applications of the affinity chromatography in drug screening.

The structure of liquid alkali nitrates and nitrites

DOE PAGES

Wilding, Martin C.; Wilson, Mark; Ribeiro, Mauro C. C.; ...

2017-07-26

State of the art high energy X-ray diffraction experiments and simulation models (employing a description of charge transfer) are applied to pure molten alkali nitrates and nitrites and uncover significant emerging structure.

Cloning, heterologous expression, and in situ characterization of the first high affinity nucleobase transporter from a protozoan.

PubMed

Burchmore, Richard J S; Wallace, Lynsey J M; Candlish, Denise; Al-Salabi, Mohammed I; Beal, Paul R; Barrett, Michael P; Baldwin, Stephen A; de Koning, Harry P

2003-06-27

While multiple nucleoside transporters, some of which can also transport nucleobases, have been cloned in recent years from many different organisms, no sequence information is available for the high affinity, nucleobase-selective transporters of metazoa, parazoa, or protozoa. We have identified a gene, TbNBT1, from Trypanosoma brucei brucei that encodes a 435-residue protein of the equilibrative nucleoside transporter superfamily. The gene was expressed in both the procyclic and bloodstream forms of the organism. Expression of TbNBT1 in a Saccharomyces cerevisiae strain lacking an endogenous purine transporter allowed growth on adenine as sole purine source and introduced a high affinity transport activity for adenine and hypoxanthine, with Km values of 2.1 +/- 0.6 and 0.66 +/- 0.22 microm, respectively, as well as high affinity for xanthine, guanine, guanosine, and allopurinol and moderate affinity for inosine. A transporter with an indistinguishable kinetic profile was identified in T. b. brucei procyclics and designated H4. RNA interference of TbNBT1 in procyclics reduced cognate mRNA levels by approximately 80% and H4 transport activity by approximately 90%. Expression of TbNBT1 in Xenopus oocytes further confirmed that this gene encodes the first high affinity nucleobase transporter from protozoa or animals to be identified at the molecular level.

Ab initio molecular dynamic study of solid-state transitions of ammonium nitrate

PubMed Central

Yu, Hongyu; Duan, Defang; Liu, Hanyu; Yang, Ting; Tian, Fubo; Bao, Kuo; Li, Da; Zhao, Zhonglong; Liu, Bingbing; Cui, Tian

2016-01-01

High-pressure polymorphism and phase transitions have wide ranging consequences on the basic properties of ammonium nitrate. However, the phase diagram of ammonium nitrate at high pressure and high temperature is still under debate. This study systematically investigates the phase transitions and structural properties of ammonium nitrate at a pressure range of 5–60 GPa and temperature range of 250–400 K by ab initio molecular dynamics simulations. Two new phases are identified: one corresponds to the experimentally observed phase IV’ and the other is named AN-X. Simultaneously, the lattice strains play a significant role in the formation and stabilization of phase IV’, providing a reasonable explanation for experimental observation of phase IV-IV’ transition which only appears under nonhydrostatic pressure. In addition, 12 O atoms neighboring the NH (N atom in ammonium cation) atom are selected as reference system to clearly display the tanglesome rotation of ammonium cation. PMID:26754622

[Effects of nitrogen application level on soil nitrate accumulation and ammonia volatilization in high-yielding wheat field].

PubMed

Wang, Dong; Yu, Zhenwen; Yu, Wenming; Shi, Yu; Zhou, Zhongxin

2006-09-01

The study showed that during the period from sowing to pre-wintering, the soil nitrate in high-yielding wheat field moved down to deeper layers, and accumulated in the layers below 140 cm. An application rate of 96-168 kg N x hm(-2) increased the nitrate content in 0-60 cm soil layer and the wheat grain yield and its protein content, and decreased the proportion of apparent N loss to applied N and the ammonia volatilization loss from basal nitrogen. Applying 240 kg N x hm(-2) promoted the downward movement of soil nitrate and its accumulation in deeper layers, increased the proportion of apparent N loss to applied N and the ammonia volatilization loss from basal nitrogen, had no significant effect on the protein content of wheat grain, but decreased the grain yield. The appropriate application rate of nitrogen on high-yielding wheat field was 132-204 kg N x hm(-2).

Bean peptides have higher in silico binding affinities than ezetimibe for the N-terminal domain of cholesterol receptor Niemann-Pick C1 Like-1.

PubMed

Real Hernandez, Luis M; Gonzalez de Mejia, Elvira

2017-04-01

Niemann-Pick C1 like-1 (NPC1L1) mediates cholesterol absorption at the apical membrane of enterocytes through a yet unknown mechanism. Bean, pea, and lentil proteins are naturally hydrolyzed during digestion to produce peptides. The potential for pulse peptides to have high binding affinities for NPC1L1 has not been determined. In this study , in silico binding affinities and interactions were determined between the N-terminal domain of NPC1L1 and 14 pulse peptides (5≥ amino acids) derived through pepsin-pancreatin digestion. Peptides were docked in triplicate to the N-terminal domain using docking program AutoDock Vina, and results were compared to those of ezetimibe, a prescribed NPC1L1 inhibitor. Three black bean peptides (-7.2 to -7.0kcal/mol) and the cowpea bean dipeptide Lys-Asp (-7.0kcal/mol) had higher binding affinities than ezetimibe (-6.6kcal/mol) for the N-terminal domain of NPC1L1. Lentil and pea peptides studied did not have high binding affinities. The common bean peptide Tyr-Ala-Ala-Ala-Thr (-7.2kcal/mol), which can be produced from black or navy bean proteins, had the highest binding affinity. Ezetimibe and peptides with high binding affinities for the N-terminal domain are expected to interact at different locations of the N-terminal domain. All high affinity black bean peptides are expected to have van der Waals interactions with SER130, PHE136, and LEU236 and a conventional hydrogen bond with GLU238 of NPC1L1. Due to their high affinity for the N-terminal domain of NPC1L1, black and cowpea bean peptides produced in the digestive track have the potential to disrupt interactions between NPC1L1 and membrane proteins that lead to cholesterol absorption. Copyright © 2017 Elsevier Inc. All rights reserved.

Algal productivity and nitrate assimilation in an effluent dominated concrete lined stream

USGS Publications Warehouse

Kent, Robert; Belitz, Kenneth; Burton, Carmen

2005-01-01

This study examined algal productivity and nitrate assimilation in a 2.85 km reach of Cucamonga Creek, California, a concrete lined channel receiving treated municipal wastewater. Stream nitrate concentrations observed at two stations indicated nearly continuous loss throughout the diel study. Nitrate loss in the reach was approximately 11 mg/L/d or 1.0 g/m2/d as N, most of which occurred during daylight. The peak rate of nitrate loss (1.13 mg/l/hr) occurred just prior to an afternoon total CO2 depletion. Gross primary productivity, as estimated by a model using the observed differences in dissolved oxygen between the two stations, was 228 mg/L/d, or 21 g/m2/d as O2. The observed diel variations in productivity, nitrate loss, pH, dissolved oxygen, and CO2indicate that nitrate loss was primarily due to algal assimilation. The observed levels of productivity and nitrate assimilation were exceptionally high on a mass per volume basis compared to studies on other streams; these rates occurred because of the shallow stream depth. This study suggests that concrete‐lined channels can provide an important environmental service: lowering of nitrate concentrations similar to rates observed in biological treatment systems.

Denitrification in a sand and gravel aquifer

USGS Publications Warehouse

Smith, R.L.; Duff, J.H.

1988-01-01

Denitrification was assayed by the acetylene blockage technique in slurried core material obtained from a freshwater sand and gravel aquifer. The aquifer, which has been contaminated with treated sewage for more than 50 years, had a contaminant plume greater than 3.5-km long. Near the contaminant source, groundwater nitrate concentrations were greater than 1 mM, whereas 0.25 km downgradient the central portion of the contaminant plume was anoxic and contained no detectable nitrate. Samples were obtained along the longitudinal axis of the plume (0 to 0.25 km) at several depths from four sites. Denitrification was evident at in situ nitrate concentrations at all sites tested; rates ranged from 2.3 to 260 pmol of N20 produced (g of wet sediment)-' h-'. Rates were highest nearest the contaminant source and decreased with increasing distance downgradient. Denitrification was the predominant nitrate-reducing activity; no evidence was found for nitrate reduction to ammonium at any site. Denitrifying activity was carbon limited and not nitrate limited, except when the ambient nitrate level was less than the detection limit, in which case, even when amended with high concentrations of glucose and nitrate, the capacity to denitrify on a short-term basis was lacking. These results demonstrate that denitrification can occur in groundwater systems and, thereby, serve as a mechanism for nitrate remoyal from groundwater.

Selection of High-Affinity Peptidic Serine Protease Inhibitors with Increased Binding Entropy from a Back-Flip Library of Peptide-Protease Fusions.

PubMed

Sørensen, Hans Peter; Xu, Peng; Jiang, Longguang; Kromann-Hansen, Tobias; Jensen, Knud J; Huang, Mingdong; Andreasen, Peter A

2015-09-25

We have developed a new concept for designing peptidic protein modulators, by recombinantly fusing the peptidic modulator, with randomized residues, directly to the target protein via a linker and screening for internal modulation of the activity of the protein. We tested the feasibility of the concept by fusing a 10-residue-long, disulfide-bond-constrained inhibitory peptide, randomized in selected positions, to the catalytic domain of the serine protease murine urokinase-type plasminogen activator. High-affinity inhibitory peptide variants were identified as those that conferred to the fusion protease the lowest activity for substrate hydrolysis. The usefulness of the strategy was demonstrated by the selection of peptidic inhibitors of murine urokinase-type plasminogen activator with a low nanomolar affinity. The high affinity could not have been predicted by rational considerations, as the high affinity was associated with a loss of polar interactions and an increased binding entropy. Copyright © 2015 Elsevier Ltd. All rights reserved.

Human llamas: adaptation to altitude in subjects with high hemoglobin oxygen affinity.

PubMed Central

Hebbel, R P; Eaton, J W; Kronenberg, R S; Zanjani, E D; Moore, L G; Berger, E M

1978-01-01

To assess the adaptive value of the right-shift of the oxyhemoglobin dissociation curve (decreased affinity for oxygen) observed in humans upon altitude exposure, the short-term physiologic responses to altitude-induced hypoxia were evaluated in two subjects with a high oxygen affinity hemoglobin (Hb Andrew-Minneapolis) and in two of their normal siblings. In striking contrast to normal subjects, at moderately high altitude (3,100 m) the high affinity subjects manifested: (a) lesser increments in resting heart rate; (b) minimal increases in plasma and urinary erythropoietin; (c) no decrement in maximal oxygen consumption; and (d) no thrombocytopenia. There was no difference between subject pairs in 2,3-diphosphoglycerate response to altitude exposure. These results tend to contradict the belief that a decrease in hemoglobin oxygen affinity is of adaptive value to humans at moderate altitudes. Rather, they support the hypothesis that, despite disadvantages at low altitude, a left-shifted oxyhemoglobin dissociation curve may confer a degree of preadaptation to altitude. PMID:29054

Genome Data Mining and Soil Survey for the Novel Group 5 [NiFe]-Hydrogenase To Explore the Diversity and Ecological Importance of Presumptive High-Affinity H2-Oxidizing Bacteria ▿†

PubMed Central

Constant, Philippe; Chowdhury, Soumitra Paul; Hesse, Laura; Pratscher, Jennifer; Conrad, Ralf

2011-01-01

Streptomyces soil isolates exhibiting the unique ability to oxidize atmospheric H2 possess genes specifying a putative high-affinity [NiFe]-hydrogenase. This study was undertaken to explore the taxonomic diversity and the ecological importance of this novel functional group. We propose to designate the genes encoding the small and large subunits of the putative high-affinity hydrogenase hhyS and hhyL, respectively. Genome data mining revealed that the hhyL gene is unevenly distributed in the phyla Actinobacteria, Proteobacteria, Chloroflexi, and Acidobacteria. The hhyL gene sequences comprised a phylogenetically distinct group, namely, the group 5 [NiFe]-hydrogenase genes. The presumptive high-affinity H2-oxidizing bacteria constituting group 5 were shown to possess a hydrogenase gene cluster, including the genes encoding auxiliary and structural components of the enzyme and four additional open reading frames (ORFs) of unknown function. A soil survey confirmed that both high-affinity H2 oxidation activity and the hhyL gene are ubiquitous. A quantitative PCR assay revealed that soil contained 106 to 108 hhyL gene copies g (dry weight)−1. Assuming one hhyL gene copy per genome, the abundance of presumptive high-affinity H2-oxidizing bacteria was higher than the maximal population size for which maintenance energy requirements would be fully supplied through the H2 oxidation activity measured in soil. Our data indicate that the abundance of the hhyL gene should not be taken as a reliable proxy for the uptake of atmospheric H2 by soil, because high-affinity H2 oxidation is a facultatively mixotrophic metabolism, and microorganisms harboring a nonfunctional group 5 [NiFe]-hydrogenase may occur. PMID:21742924

Evolution of substrate specificity for the bile salt transporter ASBT (SLC10A2)[S

PubMed Central

Lionarons, Daniël A.; Boyer, James L.; Cai, Shi-Ying

2012-01-01

The apical Na+-dependent bile salt transporter (ASBT/SLC10A2) is essential for maintaining the enterohepatic circulation of bile salts. It is not known when Slc10a2 evolved as a bile salt transporter or how it adapted to substantial changes in bile salt structure during evolution. We characterized ASBT orthologs from two primitive vertebrates, the lamprey that utilizes early 5α-bile alcohols and the skate that utilizes structurally different 5β-bile alcohols, and compared substrate specificity with ASBT from humans who utilize modern 5β-bile acids. Everted gut sacs of skate but not the more primitive lamprey transported 3H-taurocholic acid (TCA), a modern 5β-bile acid. However, molecular cloning identified ASBT orthologs from both species. Cell-based assays using recombinant ASBT/Asbt's indicate that lamprey Asbt has high affinity for 5α-bile alcohols, low affinity for 5β-bile alcohols, and lacks affinity for TCA, whereas skate Asbt showed high affinity for 5α- and 5β-bile alcohols but low affinity for TCA. In contrast, human ASBT demonstrated high affinity for all three bile salt types. These findings suggest that ASBT evolved from the earliest vertebrates by gaining affinity for modern bile salts while retaining affinity for older bile salts. Also, our results indicate that the bile salt enterohepatic circulation is conserved throughout vertebrate evolution. PMID:22669917

Root-type-specific plasticity in response to localized high nitrate supply in maize (Zea mays).

PubMed

Yu, Peng; Hochholdinger, Frank; Li, Chunjian

2015-10-01

Shoot-borne roots contribute to most of the nutrient uptake throughout the life cycle of maize (Zea mays). Compared with numerous studies with embryonic roots, detailed information on the phenotypic plasticity of shoot-borne roots in response to a heterogeneous nitrogen supply is scarce. The present study therefore provides a comprehensive profile of fine-scale plastic responses of distinct root types to localized high nitrate supply. Seedlings of the maize inbred line B73 were grown in split-root systems. The anatomy and morphological plasticity of the primary root and the roots initiated from the 2nd, 5th and 7th shoot nodes, and their lateral roots, were studied in response to local high nitrate supply to one side of the root system. In contrast to the insensitivity of axial roots, local high nitrate supply increased the length of 1st-order lateral roots on the primary root and the three whorls of shoot-borne roots at different growth stages, and increased the density of 1st-order lateral roots on the 7th shoot-borne root after silking. The length and density of 2nd-order lateral roots on the three whorls of shoot-borne roots displayed a more flexible response to local high nitrate than 1st-order lateral roots. Root diameter and number, and total area and diameter of metaxylem vessels increased from the primary root to early and then later developed shoot-borne roots, which showed a positive relationship with shoot growth and N accumulation. Maize axial roots and lateral roots responded differently to local high nitrate, and this was related to their function. The extent of morphological plasticity of lateral roots in response to local high nitrate depended on the initiation time of the shoot-borne roots on which the lateral roots developed. Morphological plasticity was higher on 2nd-order than on 1st-order lateral roots. The results suggest that higher order lateral root branching might be a potential target for genetic improvement in future maize breeding. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Hemoglobin–oxygen affinity in high-altitude vertebrates: is there evidence for an adaptive trend?

PubMed Central

2016-01-01

ABSTRACT In air-breathing vertebrates at high altitude, fine-tuned adjustments in hemoglobin (Hb)–O2 affinity provide an energetically efficient means of mitigating the effects of arterial hypoxemia. However, it is not always clear whether an increased or decreased Hb–O2 affinity should be expected to improve tissue O2 delivery under different degrees of hypoxia, due to the inherent trade-off between arterial O2 loading and peripheral O2 unloading. Theoretical results indicate that the optimal Hb–O2 affinity varies as a non-linear function of environmental O2 availability, and the threshold elevation at which an increased Hb–O2 affinity becomes advantageous depends on the magnitude of diffusion limitation (the extent to which O2 equilibration at the blood–gas interface is limited by the kinetics of O2 exchange). This body of theory provides a framework for interpreting the possible adaptive significance of evolved changes in Hb–O2 affinity in vertebrates that have colonized high-altitude environments. To evaluate the evidence for an empirical generalization and to test theoretical predictions, I synthesized comparative data in a phylogenetic framework to assess the strength of the relationship between Hb–O2 affinity and native elevation in mammals and birds. Evidence for a general trend in mammals is equivocal, but there is a remarkably strong positive relationship between Hb–O2 affinity and native elevation in birds. Evolved changes in Hb function in high-altitude birds provide one of the most compelling examples of convergent biochemical adaptation in vertebrates. PMID:27802149

Hemoglobin-oxygen affinity in high-altitude vertebrates: is there evidence for an adaptive trend?

PubMed

Storz, Jay F

2016-10-15

In air-breathing vertebrates at high altitude, fine-tuned adjustments in hemoglobin (Hb)-O 2 affinity provide an energetically efficient means of mitigating the effects of arterial hypoxemia. However, it is not always clear whether an increased or decreased Hb-O 2 affinity should be expected to improve tissue O 2 delivery under different degrees of hypoxia, due to the inherent trade-off between arterial O 2 loading and peripheral O 2 unloading. Theoretical results indicate that the optimal Hb-O 2 affinity varies as a non-linear function of environmental O 2 availability, and the threshold elevation at which an increased Hb-O 2 affinity becomes advantageous depends on the magnitude of diffusion limitation (the extent to which O 2 equilibration at the blood-gas interface is limited by the kinetics of O 2 exchange). This body of theory provides a framework for interpreting the possible adaptive significance of evolved changes in Hb-O 2 affinity in vertebrates that have colonized high-altitude environments. To evaluate the evidence for an empirical generalization and to test theoretical predictions, I synthesized comparative data in a phylogenetic framework to assess the strength of the relationship between Hb-O 2 affinity and native elevation in mammals and birds. Evidence for a general trend in mammals is equivocal, but there is a remarkably strong positive relationship between Hb-O 2 affinity and native elevation in birds. Evolved changes in Hb function in high-altitude birds provide one of the most compelling examples of convergent biochemical adaptation in vertebrates. © 2016. Published by The Company of Biologists Ltd.

Specificity of cell–cell adhesion by classical cadherins: Critical role for low-affinity dimerization through β-strand swapping

PubMed Central

Chen, Chien Peter; Posy, Shoshana; Ben-Shaul, Avinoam; Shapiro, Lawrence; Honig, Barry H.

2005-01-01

Cadherins constitute a family of cell-surface proteins that mediate intercellular adhesion through the association of protomers presented from juxtaposed cells. Differential cadherin expression leads to highly specific intercellular interactions in vivo. This cell–cell specificity is difficult to understand at the molecular level because individual cadherins within a given subfamily are highly similar to each other both in sequence and structure, and they dimerize with remarkably low binding affinities. Here, we provide a molecular model that accounts for these apparently contradictory observations. The model is based in part on the fact that cadherins bind to one another by “swapping” the N-terminal β-strands of their adhesive domains. An inherent feature of strand swapping (or, more generally, the domain swapping phenomenon) is that “closed” monomeric conformations act as competitive inhibitors of dimer formation, thus lowering affinities even when the dimer interface has the characteristics of high-affinity complexes. The model describes quantitatively how small affinity differences between low-affinity cadherin dimers are amplified by multiple cadherin interactions to establish large specificity effects at the cellular level. It is shown that cellular specificity would not be observed if cadherins bound with high affinities, thus emphasizing the crucial role of strand swapping in cell–cell adhesion. Numerical estimates demonstrate that the strength of cellular adhesion is extremely sensitive to the concentration of cadherins expressed at the cell surface. We suggest that the domain swapping mechanism is used by a variety of cell-adhesion proteins and that related mechanisms to control affinity and specificity are exploited in other systems. PMID:15937105

DNA aptamers for the detection of Haemophilus influenzae type b by cell SELEX.

PubMed

Bitaraf, F S; Rasooli, I; Mousavi Gargari, S L

2016-03-01

Haemophilus influenzae type b (Hib) causes acute bacterial meningitis (ABM) in children, with a mortality rate of about 3-6 % of the affected patients. ABM can lead to death during a period of hours to several days and, hence, rapid and early detection of the infection is crucial. Aptamers, the short single-stranded DNA or RNA with high affinity to target molecules, are selected by a high-flux screening technique known as in vitro screening and systematic evolution of ligands by exponential enrichment technology (SELEX). In this study, whole-cell SELEX was applied for the selection of target-specific aptamers with high affinity to Hib. ssDNA aptamers prepared by lambda exonuclease were incubated with the target cells (Hib). The aptameric binding rate to Hib was characterized for binding affinity after seven SELEX rounds by flow cytometry. The aptamers with higher binding affinity were cloned. Four of 68 aptamer clones were selected for sequencing. The dissociation constant (Kd) of the high-affinity aptamer clones 45 and 63 were 47.10 and 28.46 pM, respectively. These aptamers did not bind to other bacterial species, including the seven meningitis-causing bacteria. They showed distinct affinity to various H. influenzae strains only. These aptamers showed the highest affinity to Hib and the lowest affinity to H. influenzae type c and to other meningitis-causing bacteria. Clone 63 could detect Hib in patients' cerebrospinal fluid (CSF) samples at 60 colony-forming units (CFU)/mL. The results indicate applicability of the aptamers for rapid and early detection of infections brought about by Hib.

Differences between high-affinity forskolin binding sites in dopamine-riche and other regions of rat brain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Poat, J.A.; Cripps, H.E.; Iversen, L.L.

1988-05-01

Forskolin labelled with (/sup 3/H) bound to high- and low-affinity sites in the rat brain. The high-affinity site was discretely located, with highest densities in the striatum, nucleus accumbens, olfactory tubercule, substantia nigra, hippocampus, and the molecular layers of the cerebellum. This site did not correlate well with the distribution of adenylate cyclase. The high-affinity striatal binding site may be associated with a stimulatory guanine nucleotide-binding protein. Thus, the number of sites was increased by the addition of Mg/sup 2 +/ and guanylyl imidodiphosphate. Cholera toxin stereotaxically injected into rat striatum increased the number of binding sites, and no furthermore » increase was noted following the subsequent addition of guanyl nucleotide. High-affinity forskolin binding sites in non-dopamine-rich brain areas (hippocampus and cerebullum) were modulated in a qualitatively different manner by guanyl nucleotides. In these areas the number of binding sites was significantly reduced by the addition of guanyl nucleotide. These results suggest that forskolin may have a potential role in identifying different functional/structural guanine nucleotide-binding proteins.« less

Comparison of plasma generated nitrogen fertilizer to conventional fertilizers ammonium nitrate and sodium nitrate for pre-emergent and seedling growth

NASA Astrophysics Data System (ADS)

Andhavarapu, A.; King, W.; Lindsay, A.; Byrns, B.; Knappe, D.; Fonteno, W.; Shannon, S.

2014-10-01

Plasma source generated nitrogen fertilizer is compared to conventional nitrogen fertilizers in water for plant growth. Root, shoot sizes, and weights are used to examine differences between plant treatment groups. With a simple coaxial structure creating a large-volume atmospheric glow discharge, a 162 MHz generator drives the air plasma. The VHF plasma source emits a steady state glow; the high drive frequency is believed to inhibit the glow-to-arc transition for non-thermal discharge generation. To create the plasma activated water (PAW) solutions used for plant treatment, the discharge is held over distilled water until a 100 ppm nitrate aqueous concentration is achieved. The discharge is used to incorporate nitrogen species into aqueous solution, which is used to fertilize radishes, marigolds, and tomatoes. In a four week experiment, these plants are watered with four different solutions: tap water, dissolved ammonium nitrate DI water, dissolved sodium nitrate DI water, and PAW. Ammonium nitrate solution has the same amount of total nitrogen as PAW; sodium nitrate solution has the same amount of nitrate as PAW. T-tests are used to determine statistical significance in plant group growth differences. PAW fertilization chemical mechanisms are presented.

Alkyl nitrate (C1-C3) depth profiles in the tropical Pacific Ocean

NASA Astrophysics Data System (ADS)

Dahl, E. E.; Yvon-Lewis, S. A.; Saltzman, E. S.

2007-01-01

This paper reports the first depth profile measurements of methyl, ethyl, isopropyl and n-propyl nitrates in the tropical Pacific Ocean. Depth profile measurements were made at 22 stations during the Project Halocarbon Air Sea Exchange cruise, in warm pool, equatorial, subequatorial, and gyre waters. The highest concentrations, up to several hundred pM of methyl nitrate, were observed in the central Pacific within 8 degrees of the equator. In general, alkyl nitrate levels were highest in the surface mixed layer, and decreased with depth below the mixed layer. The spatial distribution of the alkyl nitrates suggests that there is a strong source associated with biologically productive ocean regions, that is characterized by high ratios of methyl:ethyl nitrate. However, the data do not allow discrimination between direct biological emissions and photochemistry as production mechanisms. Alkyl nitrates were consistently detectable at several hundred meters depth. On the basis of the estimated chemical loss rate of these compounds, we conclude that deep water alkyl nitrates must be produced in situ. Possible sources include free radical processes initiated by radioactive decay or cosmic rays, enzymatically mediated reactions involving bacteria, or unidentified chemical mechanisms involving dissolved organic matter.

Protein Tyrosine Nitration during Development and Abiotic Stress Response in Plants

PubMed Central

Mata-Pérez, Capilla; Begara-Morales, Juan C.; Chaki, Mounira; Sánchez-Calvo, Beatriz; Valderrama, Raquel; Padilla, María N.; Corpas, Francisco J.; Barroso, Juan B.

2016-01-01

In recent years, the study of nitric oxide (NO) in plant systems has attracted the attention of many researchers. A growing number of investigations have shown the significance of NO as a signal molecule or as a molecule involved in the response against (a)biotic processes. NO can be responsible of the post-translational modifications (NO-PTM) of target proteins by mechanisms such as the nitration of tyrosine residues. The study of protein tyrosine nitration during development and under biotic and adverse environmental conditions has increased in the last decade; nevertheless, there is also an endogenous nitration which seems to have regulatory functions. Moreover, the advance in proteome techniques has enabled the identification of new nitrated proteins, showing the high variability among plant organs, development stage and species. Finally, it may be important to discern between a widespread protein nitration because of greater RNS content, and the specific nitration of key targets which could affect cell-signaling processes. In view of the above point, we present a mini-review that offers an update about the endogenous protein tyrosine nitration, during plant development and under several abiotic stress conditions. PMID:27895655

Modeling of nitrate concentration in groundwater using artificial intelligence approach--a case study of Gaza coastal aquifer.

PubMed

Alagha, Jawad S; Said, Md Azlin Md; Mogheir, Yunes

2014-01-01

Nitrate concentration in groundwater is influenced by complex and interrelated variables, leading to great difficulty during the modeling process. The objectives of this study are (1) to evaluate the performance of two artificial intelligence (AI) techniques, namely artificial neural networks and support vector machine, in modeling groundwater nitrate concentration using scant input data, as well as (2) to assess the effect of data clustering as a pre-modeling technique on the developed models' performance. The AI models were developed using data from 22 municipal wells of the Gaza coastal aquifer in Palestine from 2000 to 2010. Results indicated high simulation performance, with the correlation coefficient and the mean average percentage error of the best model reaching 0.996 and 7 %, respectively. The variables that strongly influenced groundwater nitrate concentration were previous nitrate concentration, groundwater recharge, and on-ground nitrogen load of each land use land cover category in the well's vicinity. The results also demonstrated the merit of performing clustering of input data prior to the application of AI models. With their high performance and simplicity, the developed AI models can be effectively utilized to assess the effects of future management scenarios on groundwater nitrate concentration, leading to more reasonable groundwater resources management and decision-making.

Ecosystem and human health impacts from increased corn production: vulnerability assessment of exposure to high nitrate concentrations in groundwater and blue baby syndrome

NASA Astrophysics Data System (ADS)

Garcia, V.; Cooter, E. J.

2013-12-01

The Renewable Fuel Standard (RFS) requires oil refiners to reach a target of 15 billion gallons of corn-based ethanol by 2022. However, there are concerns that the broad-scale use of corn as a source of ethanol may lead to unintended economic and environmental consequences. This study applies the geophysical relationships captured with linked meteorological, air quality and agriculture models to examine the impact of corn production before enactment of the RFS in 2002 and at the height of the RFS targets in 2022. In particular, we investigate the probability of high-levels of nitrate in groundwater resulting from increased corn production and then relate this vulnerability to the potential for infants to acquire Methemoglobinemia, or 'Blue Baby Syndrome'. Blue Baby Syndrome (BBS) is a potentially fatal condition that occurs when the hemoglobin (Fe2+) in an infant's red blood cells is oxidized to methemoglobin (Fe3+), preventing the uptake of oxygen from the baby's blood. Exposure to high levels of nitrate in groundwater occur near the intersection of areas where surface water can more readily leach into shallow aquifers, wells are the main source of drinking water, and high nitrogen inputs exist. We use a coupled meteorological, agricultural and air quality model to identify areas vulnerable to increased nitrate contamination and associated risk to acquiring BBS. We first verify the relationship between predictive variables (e.g., nitrogen deposition and fertilization rates, landcover, soils and aquifer type) and nitrate groundwater levels by applying a regression model to over 800 nitrate measurements taken from wells located throughout the US (Figure 1). We then apply the regression coefficients to the coupled model output to identify areas that are at an increased risk for high nitrate groundwater levels in 2022. Finally, we examine the potential change in risk for acquiring BBS resulting from increased corn production by applying an Oral Reference Dose (RfD) factor from the US EPA Integrated Risk Information System.

Genotoxic effects of old landfill leachate on HepG2 cells after nitration/ultrafiltration/reverse osmosis membrane treatment process.

PubMed

Cheng, Rong; Zhao, Ling; Yin, Pinghe

2017-12-01

Toxicity assessment of nitration/ultrafiltration/reverse osmosis (nitration/UF/RO) project, which has recently been widely used as an efficient process with applications in practical leachate treatment, was very limited. In the present study, DNA damage of leachates was investigated before and after the nitration/UF/RO process by a battery of assays with human hepatoma cells. Methyletrazolium assay showed a high cytotoxicity of 97.1% after being exposed to the highest concentration of raw leachate for 24 h, and a cytotoxicity of 26% in effluent at a concentration of 30% (v/v). Both comet assay (24 h) and γH2AX flow cytometer assay (3 h) showed increased levels of DNA damage in cells exposed to raw leachate and after nitration/UF-treated leachate followed by a significant increase of 7-ethoxyresorufin-O-deethylase activity. However, the effluent after nitration/UF/RO treatment showed no significant difference compared to negative control for γH2AX flow cytometer assay but slight DNA damage at concentrations of 20% and 30% (v/v) as well as increase of 7-ethoxyresorufin-O-deethylase. Analysis showed that nitration/UF/RO process exhibited high removal of physicochemical indexes and significant reduction of toxic and genotoxic effects of leachate, but still demands an improvement to reduce all possible negative risks to the environment and humans. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

Nitrates for stable angina: a systematic review and meta-analysis of randomized clinical trials.

PubMed

Wei, Jiafu; Wu, Taixiang; Yang, Qing; Chen, Mao; Ni, Juan; Huang, Dejia

2011-01-07

To assess the effect (harms and benefits) of nitrates for stable angina. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE. Randomized controlled trials with both parallel and crossover design were included. The following outcome measures were evaluated: number of angina attacks weekly and nitroglycerin consumption, quality of life, total exercise duration, time to onset of angina and time to 1 mm ST depression. Fifty-one trials with 3595 patients meeting inclusion criteria were analyzed. Both intermittent and continuous regimens of nitrates lengthened exercise duration significantly by 31 and 53 s respectively. The number of angina attacks was significantly reduced by 2.89 episodes weekly for continuous administration and 1.5 episodes weekly for intermittent administration. With intermittent administration, increased dose provided with 21 s more length of exercise duration. With continuous administration, exercise duration was pronged more in low-dose group. Quality of life was not improved by continuous application of GTN patches and was similar between continuous and intermittent groups. In addition, 51.6% patients receiving nitrates complained with headache. Long-term administration of nitrates was beneficial for angina prophylaxis and improved exercise performance but might be ineffective for improving quality of life. With continuous regimen, low-dose nitrates were more effective than high-dose ones for improving exercise performance. By contrast, with intermittent regimen, high-dose nitrates were more effective. In addition, intermittent administration could bring zero-hour effect. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

Characterization of organic nitrate constituents of secondary organic aerosol (SOA) from nitrate-radical-initiated oxidation of limonene using high-resolution chemical ionization mass spectrometry

NASA Astrophysics Data System (ADS)

Faxon, Cameron; Hammes, Julia; Le Breton, Michael; Kant Pathak, Ravi; Hallquist, Mattias

2018-04-01

The gas-phase nitrate radical (NO3⚫) initiated oxidation of limonene can produce organic nitrate species with varying physical properties. Low-volatility products can contribute to secondary organic aerosol (SOA) formation and organic nitrates may serve as a NOx reservoir, which could be especially important in regions with high biogenic emissions. This work presents the measurement results from flow reactor studies on the reaction of NO3⚫ with limonene using a High-Resolution Time-of-Flight Chemical Ionization Mass Spectrometer (HR-ToF-CIMS) combined with a Filter Inlet for Gases and AEROsols (FIGAERO). Major condensed-phase species were compared to those in the Master Chemical Mechanism (MCM) limonene mechanism, and many non-listed species were identified. The volatility properties of the most prevalent organic nitrates in the produced SOA were determined. Analysis of multiple experiments resulted in the identification of several dominant species (including C10H15NO6, C10H17NO6, C8H11NO6, C10H17NO7, and C9H13NO7) that occurred in the SOA under all conditions considered. Additionally, the formation of dimers was consistently observed and these species resided almost completely in the particle phase. The identities of these species are discussed, and formation mechanisms are proposed. Cluster analysis of the desorption temperatures corresponding to the analyzed particle-phase species yielded at least five distinct groupings based on a combination of molecular weight and desorption profile. Overall, the results indicate that the oxidation of limonene by NO3⚫ produces a complex mixture of highly oxygenated monomer and dimer products that contribute to SOA formation.

Nitroproteins in Human Astrocytomas Discovered by Gel Electrophoresis and Tandem Mass Spectrometry

NASA Astrophysics Data System (ADS)

Peng, Fang; Li, Jianglin; Guo, Tianyao; Yang, Haiyan; Li, Maoyu; Sang, Shushan; Li, Xuejun; Desiderio, Dominic M.; Zhan, Xianquan

2015-12-01

Protein tyrosine nitration is involved in the pathogenesis of highly fatal astrocytomas, a type of brain cancer. To understand the molecular mechanisms of astrocytomas and to discover new biomarkers/therapeutic targets, we sought to identify nitroproteins in human astrocytoma tissue. Anti-nitrotyrosine immunoreaction-positive proteins from a high-grade astrocytoma tissue were detected with two-dimensional gel electrophoresis (2DGE)-based nitrotyrosine immunoblots, and identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fifty-seven nitrotyrosine immunopositive protein spots were detected. A total of 870 proteins (nitrated and non-nitrated) in nitrotyrosine-immunopositive 2D gel spots were identified, and 18 nitroproteins and their 20 nitrotyrosine sites were identified with MS/MS analysis. These nitroproteins participate in multiple processes, including drug-resistance, signal transduction, cytoskeleton, transcription and translation, cell proliferation and apoptosis, immune response, phenotypic dedifferentiation, cell migration, and metastasis. Among those nitroproteins that might play a role in astrocytomas was nitro-sorcin, which is involved in drug resistance and metastasis and might play a role in the spread and treatment of an astrocytoma. Semiquantitative immune-based measurements of different sorcin expressions were found among different grades of astrocytomas relative to controls, and a semiquantitative increased nitration level in high-grade astrocytoma relative to control. Nitro-β-tubulin functions in cytoskeleton and cell migration. Semiquantitative immunoreactivity of β-tubulin showed increased expression among different grades of astrocytomas relative to controls and semiquantitatively increased nitration level in high-grade astrocytoma relative to control. Each nitroprotein was rationalized and related to the corresponding functional system to provide new insights into tyrosine nitration and its potential role in the pathogenesis of astrocytoma formation.

Nitrate-Containing Beetroot Juice Reduces Oxygen Consumption During Submaximal Exercise in Low but Not High Aerobically Fit Male Runners.

PubMed

Carriker, Colin R; Vaughan, Roger A; VanDusseldorp, Trisha A; Johnson, Kelly E; Beltz, Nicholas M; McCormick, James J; Cole, Nathan H; Gibson, Ann L

2016-12-31

to examine the effect of a 4-day NO3- loading protocol on the submaximal oxygen cost of both low fit and high fit participants at five different exercise intensities. participants were initially assigned to a placebo (PL; negligible NO3-) or inorganic nitrate-rich (NR; 6.2 mmol nitrate/day) group; double-blind, placebo-controlled, crossover. Participants completed three trials (T1, T2 and T3). T1 included a maximal aerobic capacity (VO2max) treadmill test. A 6-day washout, minimizing nitrate consumption, preceded T2. Each of the four days prior to T2 and T3, participants consumed either PL or NR; final dose 2.5 hours prior to exercise. A 14-day washout followed T2. T2 and T3 consisted of 5-minute submaximal treadmill bouts (45, 60, 70, 80 and 85% VO2max) determined during T1. Low fit nitrate-supplemented participants consumed less oxygen (p<0.05) at lower workloads (45% and 60% VO2max) compared to placebo trials; changes not observed in high fit participants. The two lowest intensity workloads of 45 and 60% VO2max revealed the greatest correlation (r=0.54, p=0.09 and r=0.79, p<0.05; respectively). No differences were found between conditions for heart rate, respiratory exchange ratio or rating of perceived exertion for either fitness group. Nitrate consumption promotes reduced oxygen consumption at lower exercise intensities in low fit, but not high fit males. Lesser fit individuals may receive greater benefit than higher fit participants exercising at intensities <60% VO2max.

SELMAP - SELEX affinity landscape MAPping of transcription factor binding sites using integrated microfluidics

PubMed Central

Chen, Dana; Orenstein, Yaron; Golodnitsky, Rada; Pellach, Michal; Avrahami, Dorit; Wachtel, Chaim; Ovadia-Shochat, Avital; Shir-Shapira, Hila; Kedmi, Adi; Juven-Gershon, Tamar; Shamir, Ron; Gerber, Doron

2016-01-01

Transcription factors (TFs) alter gene expression in response to changes in the environment through sequence-specific interactions with the DNA. These interactions are best portrayed as a landscape of TF binding affinities. Current methods to study sequence-specific binding preferences suffer from limited dynamic range, sequence bias, lack of specificity and limited throughput. We have developed a microfluidic-based device for SELEX Affinity Landscape MAPping (SELMAP) of TF binding, which allows high-throughput measurement of 16 proteins in parallel. We used it to measure the relative affinities of Pho4, AtERF2 and Btd full-length proteins to millions of different DNA binding sites, and detected both high and low-affinity interactions in equilibrium conditions, generating a comprehensive landscape of the relative TF affinities to all possible DNA 6-mers, and even DNA10-mers with increased sequencing depth. Low quantities of both the TFs and DNA oligomers were sufficient for obtaining high-quality results, significantly reducing experimental costs. SELMAP allows in-depth screening of hundreds of TFs, and provides a means for better understanding of the regulatory processes that govern gene expression. PMID:27628341

Tracing the Fate of Atmospheric Nitrate in a Subalpine Watershed Using Δ17O.

PubMed

Bourgeois, Ilann; Savarino, Joël; Caillon, Nicolas; Angot, Hélène; Barbero, Albane; Delbart, Franck; Voisin, Didier; Clément, Jean-Christophe

2018-05-15

Nitrogen is an essential nutrient for life on Earth, but in excess, it can lead to environmental issues (e.g., N saturation, loss of biodiversity, acidification of lakes, etc.). Understanding the nitrogen budget (i.e., inputs and outputs) is essential to evaluate the prospective decay of the ecosystem services (e.g., freshwater quality, erosion control, loss of high patrimonial-value plant species, etc.) that subalpine headwater catchments provide, especially as these ecosystems experience high atmospheric nitrogen deposition. Here, we use a multi-isotopic tracer (Δ 17 O, δ 15 N and δ 18 O) of nitrate in aerosols, snow, and streams to assess the fate of atmospherically deposited nitrate in the subalpine watershed of the Lautaret Pass (French Alps). We show that atmospheric N deposition contributes significantly to stream nitrate pool year-round, either by direct inputs (up to 35%) or by in situ nitrification of atmospheric ammonium (up to 35%). Snowmelt in particular leads to high exports of atmospheric nitrate, most likely fast enough to impede assimilation by surrounding ecosystems. Yet, in a context of climate change, with shorter snow seasons, and increasing nitrogen emissions, our results hint at possibly stronger ecological consequences of nitrogen atmospheric deposition in the close future.

Nitrate Reductase Activity and Polyribosomal Content of Corn (Zea mays L.) Having Low Leaf Water Potentials 1

PubMed Central

Morilla, Camila A.; Boyer, J. S.; Hageman, R. H.

1973-01-01

Desiccation of 8- to 13-day-old seedlings, achieved by withholding nutrient solution from the vermiculite root medium, caused a reduction in nitrate reductase activity of the leaf tissue. Activity declined when leaf water potentials decreased below −2 bars and was 25% of the control at a leaf water potential of −13 bars. Experiments were conducted to determine whether the decrease in nitrate reductase activity was due to reduced levels of nitrate in the tissue, direct inactivation of the enzyme by low leaf water potentials, or to changes in rates of synthesis or decay of the enzyme. Although tissue nitrate content decreased with the onset of desiccation, it did not continue to decline with tissue desiccation and loss of enzyme activity. Nitrate reductase activity recovered when the plants were rewatered with nitrate-free medium, suggesting that the nitrate in the plant was adequate for high nitrate reductase activity. The rate of decay of nitrate reductase activity from desiccated tissue was essentially identical to that of the control, in vivo or in vitro, regardless of the rapidity of desiccation of the tissue. Direct inactivation of the enzyme by the low water potentials was not detected. Polyribosomal content of the tissue declined with the decrease in water potential, prior to the decline in nitrate reductase activity. Changes in ribosomal profiles occurred during desiccation, regardless of whether the tissue had been excised or not and whether desiccation was rapid or slow. Reduction in polyribosomal content did not appear to be associated with changes in ribonuclease activity. Nitrate reductase activity and the polyribosomal content of the tissue recovered upon rewatering, following the recovery in water potential. The increase in polyribosomal content preceded the increase in nitrate reductase activity. Recovery of enzyme activity was prevented by cycloheximide. Based on these results, it appears that nitrate reductase activity was affected primarily by a decrease in the rate of enzyme synthesis at low leaf water potentials. PMID:16658419

The atmospheric chemistry of isoprene- and other multifunctional-nitrates

NASA Astrophysics Data System (ADS)

Perring, Anne Elizabeth

Formation of alkyl and multifunctional nitrates significantly reduces ozone production rates in their source regions, their transport and subsequent chemistry can impact secondary organic aerosol formation and NOy removal rates and they may lead to the re-release of NOx in regions far-removed from the original source. In this dissertation, the atmospheric chemistry of alkyl and multifunctional nitrates is investigated through a combination of laboratory and field measurements. In contrast to many previous studies that have focused on observations of specific individual nitrate compounds, the work described here uses a technique (Thermal Dissociation-Laser Induced Fluorescence or TD-LIF) that allows for measurements of the sum of all alkyl and multifunctional nitrates. These observations show that alkyl and multifunctional nitrates are a significant fraction of NOy in a number of different chemical regimes representing diverse hydrocarbon mixtures. In what follows, I show that their formation impacts both ozone 1 formation and NOy transport in ways that are not accounted for by currently accepted chemical mechanisms. Aircraft measurements are used to constrain nitrate yields following isoprene oxidation by OH, the atmospheric lifetimes of these nitrates, and the retention rate of the nitrate functional group upon oxidation of the initial isoprene nitrates. It is found that nitrate functionality is maintained upon further oxidation at least 75% of the time indicating that the lifetime of isoprene nitrates as a pool of compounds is considerably longer than the lifetime of the individual isoprene nitrates with respect to reaction with OH. We examine the products of NO3-initiated oxidation of isoprene in a smog-chamber, propose a detailed reaction scheme, and find that nitrates are produced with a yield of 65+/-12%, the majority of which are carbonyl nitrates. We investigate the role of alkyl and multifunctional nitrates in the Mexico City plume where they are observed to be more abundant than expected given currently accepted chemical mechanisms. We investigate several possible explanations for the unexpectedly high abundances and calculate that their formation depresses ozone production in the near-field of Mexico City by as much as 30%.

Thermodynamic modeling of Cl(-), NO3(-) and SO4(2-) removal by an anion exchange resin and comparison with Dubinin-Astakhov isotherms.

PubMed

Dron, Julien; Dodi, Alain

2011-03-15

The removal of chloride, nitrate, and sulfate ions from wastewaters by a macroporous ion-exchange resin is studied through the experimental results obtained for six ion exchange systems, OH(-)/Cl(-), OH(-)/NO3(-), OH(-)/SO4(2-), and HCO3(-)/Cl(-), Cl(-)/NO3(-), Cl(-)/SO4(2-). The results are described through thermodynamic modeling, considering either an ideal or a nonideal behavior of the ionic species in the liquid and solid phases. The nonidealities are determined by the Davies equation and Wilson equations in the liquid and solid phases, respectively. The results show that the resin has a strong affinity for all the target ions, and the order of affinity obtained is OH(-) < HCO3(-) < Cl(-) < NO3(-) < SO4(2-). The calculation of the changes in standard Gibbs free energies (ΔG(0)) shows that even though HCO3(-) has a lower affinity to the resin, it may affect the removal of Cl(-), and in the same way that Cl(-) may affect the removal of NO3(-) and SO4(2-). The application of nonidealities in the thermodynamic model leads to an improved fit of the model to the experimental data with average relative deviations below 1.5% except for the OH(-)/SO4(2-) system. On the other hand, considering ideal or nonideal behaviors has no significant impact on the determination of the selectivity coefficients. The thermodynamic modeling is also compared with the Dubinin-Astakhov adsorption isotherms obtained for the same ion exchange systems. Surprisingly, the latter performs significantly better than the ideal thermodynamic model and nearly as well as the nonideal thermodynamic model.

Estimating ammonium and nitrate load from septic systems to surface water bodies within ArcGIS environments

NASA Astrophysics Data System (ADS)

Zhu, Yan; Ye, Ming; Roeder, Eberhard; Hicks, Richard W.; Shi, Liangsheng; Yang, Jinzhong

2016-01-01

This paper presents a recently developed software, ArcGIS-based Nitrogen Load Estimation Toolkit (ArcNLET), for estimating nitrogen loading from septic systems to surface water bodies. The load estimation is important for managing nitrogen pollution, a world-wide challenge to water resources and environmental management. ArcNLET simulates coupled transport of ammonium and nitrate in both vadose zone and groundwater. This is a unique feature that cannot be found in other ArcGIS-based software for nitrogen modeling. ArcNLET is designed to be flexible for the following four simulating scenarios: (1) nitrate transport alone in groundwater; (2) ammonium and nitrate transport in groundwater; (3) ammonium and nitrate transport in vadose zone; and (4) ammonium and nitrate transport in both vadose zone and groundwater. With this flexibility, ArcNLET can be used as an efficient screening tool in a wide range of management projects related to nitrogen pollution. From the modeling perspective, this paper shows that in areas with high water table (e.g. river and lake shores), it may not be correct to assume a completed nitrification process that converts all ammonium to nitrate in the vadose zone, because observation data can indicate that substantial amount of ammonium enters groundwater. Therefore, in areas with high water table, simulating ammonium transport and estimating ammonium loading, in addition to nitrate transport and loading, are important for avoiding underestimation of nitrogen loading. This is demonstrated in the Eggleston Heights neighborhood in the City of Jacksonville, FL, USA, where monitoring well observations included a well with predominant ammonium concentrations. The ammonium loading given by the calibrated ArcNLET model can be 10-18% of the total nitrogen load, depending on various factors discussed in the paper.

Multi-year record of atmospheric and snow surface nitrate in the central Antarctic plateau.

PubMed

Traversi, R; Becagli, S; Brogioni, M; Caiazzo, L; Ciardini, V; Giardi, F; Legrand, M; Macelloni, G; Petkov, B; Preunkert, S; Scarchilli, C; Severi, M; Vitale, V; Udisti, R

2017-04-01

Continuous all year-round samplings of atmospheric aerosol and surface snow at high (daily to 4-day) resolution were carried out at Dome C since 2004-05 to 2013 and nitrate records are here presented. Basing on a larger statistical data set than previous studies, results confirm that nitrate seasonal pattern is characterized by maxima during austral summer for both aerosol and surface snow, occurring in-phase with solar UV irradiance. This temporal pattern is likely due to a combination of nitrate sources and post-depositional processes whose intensity usually enhances during the summer. Moreover, it should be noted that a case study of the synoptic conditions, which took place during a major nitrate event, showed the occurrence of a stratosphere-troposphere exchange. The sampling of both matrices at the same time with high resolution allowed the detection of a an about one-month long recurring lag of summer maxima in snow with respect to aerosol. This result can be explained by deposition and post-deposition processes occurring at the atmosphere-snow interface, such as a net uptake of gaseous nitric acid and a replenishment of the uppermost surface layers driven by a larger temperature gradient in summer. This hypothesis was preliminarily tested by a comparison with surface layers temperature data in the 2012-13 period. The analysis of the relationship between the nitrate concentration in the gas phase and total nitrate obtained at Dome C (2012-13) showed the major role of gaseous HNO 3 to the total nitrate budget suggesting the need to further investigate the gas-to-particle conversion processes. Copyright © 2017 Elsevier Ltd. All rights reserved.

The Nitrate Inventory of Unsaturated Soils at the Barrow Environmental Observatory: Current Conditions and Potential Future Trajectories

NASA Astrophysics Data System (ADS)

Heikoop, J. M.; Newman, B. D.; Arendt, C. A.; Andresen, C. G.; Lara, M. J.; Wainwright, H. M.; Throckmorton, H.; Graham, D. E.; Wilson, C. J.; Wullschleger, S. D.; Romanovsky, V. E.; Bolton, W. R.; Wales, N. A.; Rowland, J. C.

2016-12-01

Studies conducted in the Barrow Environmental Observatory under the auspices of the United States Department of Energy Next Generation Ecosystem Experiment (NGEE) - Arctic have demonstrated measurable nitrate concentrations ranging from <1 to 17 mg/L in the unsaturated centers of high-centered polygons. Conversely, nitrate concentrations in saturated areas of polygonal terrain were generally below the limit of detection. Isotopic analysis of this nitrate demonstrates that it results from microbial nitrification. The study site currently comprises mostly saturated soils. Several factors, however, could lead to drying of soils on different time scales. These include 1) topographic inversion of polygonal terrain associated with ice-wedge degradation, 2) increased connectivity and drainage of polygon troughs, similarly related to the thawing and subsidence of ice-wedges, and 3) near-surface soil drainage associated with wide-spread permafrost thaw and active layer deepening. Using a GIS approach we will estimate the current inventory of nitrate in the NGEE intensive study site using soil moisture data and existing unsaturated zone nitrate concentration data and new concentration data collected in the summer of 2016 from high- and flat-centered polygons and the elevated rims of low-centered polygons. Using this baseline, we will present potential future inventories based on various scenarios of active layer thickening and landscape geomorphic reorganization associated with permafrost thaw. Predicted inventories will be based solely on active layer moisture changes, ignoring for now potential changes associated with mineralization and nitrification of previously frozen old organic matter and changes in vegetation communities. We wish to demonstrate that physical landscape changes alone could have a profound effect on future nitrate availability. Nitrate data from recent NGEE campaigns in the Seward Peninsula of Alaska will also be presented.

Characterization of nicotine binding to the rat brain P/sub 2/ preparation: the identification of multiple binding sites which include specific up-regulatory site(s)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sloan, J.W.

1984-01-01

These studies show that nicotine binds to the rat brain P/sub 2/ preparation by saturable and reversible processes. Multiple binding sites were revealed by the configuration of saturation, kinetic and Scatchard plots. A least squares best fit of Scatchard data using nonlinear curve fitting programs confirmed the presence of a very high affinity site, an up-regulatory site, a high affinity site and one or two low affinity sites. Stereospecificity was demonstrated for the up-regulatory site where (+)-nicotine was more effective and for the high affinity site where (-)-nicotine had a higher affinity. Drugs which selectively up-regulate nicotine binding site(s) havemore » been identified. Further, separate very high and high affinity sites were identified for (-)- and (+)-(/sup 3/H)nicotine, based on evidence that the site density for the (-)-isomer is 10 times greater than that for the (+)-isomer at these sites. Enhanced nicotine binding has been shown to be a statistically significant phenomenon which appears to be a consequence of drugs binding to specific site(s) which up-regulate binding at other site(s). Although Scatchard and Hill plots indicate positive cooperatively, up-regulation more adequately describes the function of these site(s). A separate up-regulatory site is suggested by the following: (1) Drugs vary markedly in their ability to up-regulate binding. (2) Both the affinity and the degree of up-regulation can be altered by structural changes in ligands. (3) Drugs with specificity for up-regulation have been identified. (4) Some drugs enhance binding in a dose-related manner. (5) Competition studies employing cold (-)- and (+)-nicotine against (-)- and (+)-(/sup 3/H)nicotine show that the isomers bind to separate sites which up-regulate binding at the (-)- and (+)-nicotine high affinity sites and in this regard (+)-nicotine is more specific and efficacious than (-)-nicotine.« less

Nitrate formation during ozonation as a surrogate parameter for abatement of micropollutants and the N-nitrosodimethylamine (NDMA) formation potential.

PubMed

Song, Yang; Breider, Florian; Ma, Jun; von Gunten, Urs

2017-10-01

In this study, nitrate formation from ammonium and/or dissolved organic nitrogen (DON) was investigated as a novel surrogate parameter to evaluate the abatement of micropollutants during ozonation of synthetic waters containing natural organic matter (NOM) isolates, a natural water and secondary wastewater effluents. Nitrate formation during ozonation was compared to the changes in UV absorbance at 254 nm (UVA 254 ) including the effect of pH. For low specific ozone doses UVA 254 was abated more efficiently than nitrate was formed. This is due to a relatively slow rate-limiting step for nitrate formation from the reaction between ozone and a proposed nitrogen-containing intermediate. This reaction cannot compete with the fast reactions between ozone and UV-absorbing moieties (e.g., activated aromatic compounds). To further test the kinetics of nitrate formation, two possible intermediates formed during ozonation of DON were tested. At pH 7, nitrate was formed during ozonation of acetone oxime and methyl nitroacetate with second-order rate constants of 256.7 ± 4.7 M -1  s -1 and 149.5 ± 5.8 M -1  s -1 , respectively. The abatement of the selected micropollutants (i.e., 17α-ethinylestradiol (EE2), carbamazepine (CBZ), bezafibrate (BZF), ibuprofen (IBU), and p-chlorobenzoic acid (pCBA)) was investigated for specific ozone doses ≤1.53 mgO 3 /mgDOC and its efficiency depended strongly on the reactivity of the selected compounds with ozone. The relative abatement of micropollutants (i.e., EE2 and CBZ) with high ozone reactivity showed linear relationships with nitrate formation. The abatement of micropollutants with intermediate-low ozone reactivity (BZF, IBU, and pCBA) followed one- and two-phase behaviors relative to nitrate formation during ozonation of water samples containing high and low concentrations of nitrate-forming DON, respectively. During ozonation of a wastewater sample, the N-nitrosodimethylamine formation potential (NDMA-FP) during chloramination decreased with increasing specific ozone doses. A good correlation was obtained between NDMA-FP abatement and nitrate formation. Therefore, nitrate formation after pre-ozonation may be a useful parameter to estimate the reduction of the NDMA-FP during post-chloramination. Overall, the results of this study suggest that nitrate formation (possibly in combination with UVA 254 abatement) during ozonation of DON-containing waters may be a good surrogate for assessing the abatement of micropollutants and the NDMA-FP. Copyright © 2017 Elsevier Ltd. All rights reserved.

Modelling nitrate from land-surface to wells-perforations under Mediterranean agricultural land: success, failure, and future scenarios

NASA Astrophysics Data System (ADS)

Levy, Yehuda; Chefetz, Benny; Shapira, Roi; Kurtzman, Daniel

2017-04-01

Contamination of groundwater resources by nitrate due to leaching under agricultural land is probably the most troublesome agriculture-related water contamination, worldwide. Deep soil sampling (10 m) were used for calibrating vertical flow and nitrogen-transport numerical models of the unsaturated zone, under different agricultural land uses. Vegetables fields (potato and strawberries) and deciduous (persimmon) orchards in the Sharon area overlaying the coastal aquifer of Israel, were examined. Average nitrate-nitrogen fluxes below vegetables fields were 210-290 kg ha-1 a-1 and under deciduous orchards were 110-140 kg ha-1 a-1. The output water and nitrate-nitrogen fluxes of the unsaturated zone models were used as input for a three dimensional flow and nitrate-transport model in the aquifer under an area of 13.3 square kilometers of agricultural land. The area was subdivided to 4 agricultural land-uses: vegetables, deciduous, citrus orchards and non-cultivated. Fluxes of water and nitrate-nitrogen below citrus orchards were taken from a previous study in this area (Kurtzman et al., 2013, j. Contam. Hydrol.). The groundwater flow model was calibrated to well heads only by changing the hydraulic conductivity while transient recharge fluxes were constraint to the bottom-fluxes of the unsaturated zone flow models. The nitrate-transport model in the aquifer, which was fed at the top by the nitrate fluxes of the unsaturated zone models, succeeded in reconstructing the average nitrate concentration in the wells. On the other hand, this transport model failed in calculating the high concentrations in the most contaminated wells and the large spatial variability of nitrate-concentrations in the aquifer. In order to reconstruct the spatial variability and enable predictions nitrate-fluxes from the unsaturated zone were multiplied by local multipliers. This action was rationalized by the fact that the high concentrations in some wells cannot be explained by regular agricultural activity, and are probably a result of some malfunction in the well area. Prediction of the nitrate concentration 40 years to the future with 3 nitrogen-fertilization scenarios showed the following: 1) under "business as usual" fertilization scenario, the NO3 concentration will increase in average by 19 mg l-1; 2) In reducing 25% of the nitrogen fertilization mass scenario, the nitrate concentration in the aquifer will stabilize; 3) In reducing 50% of the nitrogen fertilization mass scenario, the concentration will decrease in average by 18 mg l-1.

Symptoms of nitrogen saturation in two central Appalachian hardwood forest ecosystems

Treesearch

William T. Peterjohn; Mary Beth Adams; Frank S. Gilliam

1996-01-01

By synthesizing more than twenty years of research at the Fernow Experimental Forest, we have documented 7 symptoms of nitrogen saturation in two adjacent watersheds. The symptoms include: 1) high relative rates of net nitrification, 2) long-term increases in streamwater concentrations of nitrate and base cations, 3) relatively high nitrate concentrations in solution...

A molecular recognizing system of serotonin in rat fetal axonal growth cones: uptake and high affinity binding.

PubMed

Mercado, R; Hernández, J

1992-09-18

Axonal growth cone particles (AGCP) isolated from prenatal and postnatal rat brain had different high-affinity 5-HT uptake characteristics. In postnatal AGCP the uptake behaves as in the adult rat brain, while in the prenatal AGCP the uptake characteristics seem to be in a transitional stage. Also in prenatal AGCP we observed specific, high-affinity 5-HT binding sites. These results support the idea of an important role for 5-HT during axogenesis.

Small scale denitrification variability in riparian zones: Results from a high-resolution dataset

NASA Astrophysics Data System (ADS)

Gassen, Niklas; Knöller, Kay; Musolff, Andreas; Popp, Felix; Lüders, Tillmann; Stumpp, Christine

2017-04-01

Riparian zones are important compartments at the interface between groundwater and surface water where biogeochemical processes like denitrification are often enhanced. Nitrate loads of either groundwater entering a stream through the riparian zone or streamwater infiltrating into the riparian zone can be substantially reduced. These processes are spatially and temporally highly variable, making it difficult to capture solute variabilities, estimate realistic turnover rates and thus to quantify integral mass removal. A crucial step towards a more detailed characterization is to monitor solutes on a scale which adequately resemble the highly heterogeneous distribution and on a scale where processes occur. We measured biogeochemical parameters in a spatial high resolution within a riparian corridor of a German lowland river system over the course of one year. Samples were taken from three newly developed high-resolution multi-level wells with a maximum vertical resolution of 5 cm and analyzed for major ions, DOC and N-O isotopes. Sediment derived during installation of the wells was analyzed for specific denitrifying enzymes. Results showed a distinct depth zonation of hydrochemistry within the shallow alluvial aquifer, with a 1 m thick zone just below the water table with lower nitrate concentrations and EC values similar to the nearby river. Conservative parameters were consistent inbetween the three wells, but nitrate was highly variable. In addition, spots with low nitrate concentrations showed isotopic and microbial evidence for higher denitrification activities. The depth zonation was observed throughout the year, with stronger temporal variations of nitrate concentrations just below the water table compared to deeper layers. Nitrate isotopes showed a clear seasonal trend of denitrification activities (high in summer, low in winter). Our dataset gives new insight into river-groundwater exchange processes and shows the highly heterogeneous distribution of denitrification in riparian zones, both in time and space. With these new insights, we are able to improve our understanding of spatial scaling of denitrification processes. This leads to a better prediction and improved management strategies for buffer mechanisms in riparian zones.

Vascular effects of dietary nitrate (as found in green leafy vegetables and beetroot) via the nitrate‐nitrite‐nitric oxide pathway

PubMed Central

Lidder, Satnam; Webb, Andrew J.

2013-01-01

The discovery that dietary (inorganic) nitrate has important vascular effects came from the relatively recent realization of the ‘nitrate‐nitrite‐nitric oxide (NO) pathway’. Dietary nitrate has been demonstrated to have a range of beneficial vascular effects, including reducing blood pressure, inhibiting platelet aggregation, preserving or improving endothelial dysfunction, enhancing exercise performance in healthy individuals and patients with peripheral arterial disease. Pre‐clinical studies with nitrate or nitrite also show the potential to protect against ischaemia‐reperfusion injury and reduce arterial stiffness, inflammation and intimal thickness. However, there is a need for good evidence for hard endpoints beyond epidemiological studies. Whilst these suggest reduction in cardiovascular risk with diets high in nitrate‐rich vegetables (such as a Mediterranean diet), others have suggested possible small positive and negative associations with dietary nitrate and cancer, but these remain unproven. Interactions with other nutrients, such as vitamin C, polyphenols and fatty acids may enhance or inhibit these effects. In order to provide simple guidance on nitrate intake from different vegetables, we have developed the Nitrate ‘Veg‐Table’ with ‘Nitrate Units’ [each unit being 1 mmol of nitrate (62 mg)] to achieve a nitrate intake that is likely to be sufficient to derive benefit, but also to minimize the risk of potential side effects from excessive ingestion, given the current available evidence. The lack of data concerning the long term effects of dietary nitrate is a limitation, and this will need to be addressed in future trials. PMID:22882425

Biogeochemistry of beetle-killed forests: Explaining a weak nitrate response

PubMed Central

Rhoades, Charles C.; McCutchan, James H.; Cooper, Leigh A.; Clow, David; Detmer, Thomas M.; Briggs, Jennifer S.; Stednick, John D.; Veblen, Thomas T.; Ertz, Rachel M.; Likens, Gene E.; Lewis, William M.

2013-01-01

A current pine beetle infestation has caused extensive mortality of lodgepole pine (Pinus contorta) in forests of Colorado and Wyoming; it is part of an unprecedented multispecies beetle outbreak extending from Mexico to Canada. In United States and European watersheds, where atmospheric deposition of inorganic N is moderate to low (<10 kg⋅ha⋅y), disturbance of forests by timber harvest or violent storms causes an increase in stream nitrate concentration that typically is close to 400% of predisturbance concentrations. In contrast, no significant increase in streamwater nitrate concentrations has occurred following extensive tree mortality caused by the mountain pine beetle in Colorado. A model of nitrate release from Colorado watersheds calibrated with field data indicates that stimulation of nitrate uptake by vegetation components unaffected by beetles accounts for significant nitrate retention in beetle-infested watersheds. The combination of low atmospheric N deposition (<10 kg⋅ha⋅y), tree mortality spread over multiple years, and high compensatory capacity associated with undisturbed residual vegetation and soils explains the ability of these beetle-infested watersheds to retain nitrate despite catastrophic mortality of the dominant canopy tree species. PMID:23319612

Environmental biogeography of near-surface phytoplankton in the southeast Pacific Ocean

NASA Astrophysics Data System (ADS)

Hardy, John; Hanneman, Andrew; Behrenfeldt, Michael; Horner, Rita

1996-10-01

Biogeographic interpretation of large-scale phytoplankton distribution patterns in relation to surface hydrography is essential to understanding pelagic food web dynamics and biogeochemical processes influencing global climate. We examined the abundance and biomass of phytoplankton in relation to physical and chemical parameters in the southeast Pacific Ocean. Samples were collected along longitude 110°W, between 10°N and 60°S during late austral summer. Patterns of taxa abundance and hydrographic variables were interpreted by principal components analysis. Five distinct phytohydrographic regions were identified: (i) a north equatorial region of moderate productivity dominated by small flagellates, low nitrate and low-to-moderate pCO 2; (ii) a south equatorial region characterized by high primary productivity dominated by diatoms, high nutrient levels, and relatively high pCO 2; (iii) a central gyre region characterized by low productivity dominated by small flagellates, low nitrate, and high pCO 2; (iv) a sub-Antarctic region with moderate productivity dominated by coccolithophores, moderate nitrate concentrations, and low pCO 2; and (v) an Antarctic region with high productivity dominated by diatoms, very high nitrate, and low pCO 2. Productivity and average phytoplankton cell size were positively correlated with nitrate concentration. Total phytoplankton abundance was negatively correlated with pCO 2, photosynthetically active radiation, and ultraviolet-B radiation. The interaction between phytoplankton carbon assimilation, atmospheric CO2, and the inhibitory effect of ultraviolet radiation could have implications for the global climate. These data suggest that the effects would be greatest at southern mid-latitudes (40-50°S) where present phytoplankton production and predicted future increases in UV-B are both relatively high.

Surfactant-free Colloidal Particles with Specific Binding Affinity

PubMed Central

2017-01-01

Colloidal particles with specific binding affinity are essential for in vivo and in vitro biosensing, targeted drug delivery, and micrometer-scale self-assembly. Key to these techniques are surface functionalizations that provide high affinities to specific target molecules. For stabilization in physiological environments, current particle coating methods rely on adsorbed surfactants. However, spontaneous desorption of these surfactants typically has an undesirable influence on lipid membranes. To address this issue and create particles for targeting molecules in lipid membranes, we present here a surfactant-free coating method that combines high binding affinity with stability at physiological conditions. After activating charge-stabilized polystyrene microparticles with EDC/Sulfo-NHS, we first coat the particles with a specific protein and subsequently covalently attach a dense layer of poly(ethyelene) glycol. This polymer layer provides colloidal stability at physiological conditions as well as antiadhesive properties, while the protein coating provides the specific affinity to the targeted molecule. We show that NeutrAvidin-functionalized particles bind specifically to biotinylated membranes and that Concanavalin A-functionalized particles bind specifically to the glycocortex of Dictyostelium discoideum cells. The affinity of the particles changes with protein density, which can be tuned during the coating procedure. The generic and surfactant-free coating method reported here transfers the high affinity and specificity of a protein onto colloidal polystyrene microparticles. PMID:28847149

Interaction of Ochratoxin A and Its Thermal Degradation Product 2'R-Ochratoxin A with Human Serum Albumin.

PubMed

Sueck, Franziska; Poór, Miklós; Faisal, Zelma; Gertzen, Christoph G W; Cramer, Benedikt; Lemli, Beáta; Kunsági-Máté, Sándor; Gohlke, Holger; Humpf, Hans-Ulrich

2018-06-22

Ochratoxin A (OTA) is a toxic secondary metabolite produced by several fungal species of the genus Penicillium and Aspergillus . 2′ R -Ochratoxin A (2′ R -OTA) is a thermal isomerization product of OTA formed during food processing at high temperatures. Both compounds are detectable in human blood in concentrations between 0.02 and 0.41 µg/L with 2′ R -OTA being only detectable in the blood of coffee drinkers. Humans have approximately a fifty-fold higher exposure through food consumption to OTA than to 2′ R -OTA. In human blood, however, the differences between the concentrations of the two compounds is, on average, only a factor of two. To understand these unexpectedly high 2′ R -OTA concentrations found in human blood, the affinity of this compound to the most abundant protein in human blood the human serum albumin (HSA) was studied and compared to that of OTA, which has a well-known high binding affinity. Using fluorescence spectroscopy, equilibrium dialysis, circular dichroism (CD), high performance affinity chromatography (HPAC), and molecular modelling experiments, the affinities of OTA and 2′ R -OTA to HSA were determined and compared with each other. For the affinity of HSA towards OTA, a log K of 7.0⁻7.6 was calculated, while for its thermally produced isomer 2′ R -OTA, a lower, but still high, log K of 6.2⁻6.4 was determined. The data of all experiments showed consistently that OTA has a higher affinity to HSA than 2′ R -OTA. Thus, differences in the affinity to HSA cannot explain the relatively high levels of 2′ R -OTA found in human blood samples.

Synthesis of microspheres of triuranium octaoxide by simultaneous water and nitrate extraction from ascorbate-uranyl sols.

PubMed

Brykala, M; Deptula, A; Rogowski, M; Lada, W; Olczak, T; Wawszczak, D; Smolinski, T; Wojtowicz, P; Modolo, G

A new method for synthesis of uranium oxide microspheres (diameter <100 μm) has been developed. It is a variant of our patented Complex Sol-Gel Process, which has been used to synthesize high-quality powders of a wide variety of complex oxides. Starting uranyl-nitrate-ascorbate sols were prepared by addition of ascorbic acid to uranyl nitrate hexahydrate solution and alkalizing by aqueous ammonium hydroxide and then emulsified in 2-ethylhexanol-1 containing 1v/o SPAN-80. Drops of emulsion were firstly gelled by extraction of water by the solvent. Destruction of the microspheres during thermal treatment, owing to highly reactive components in the gels, requires modification of the gelation step by Double Extraction Process-simultaneously extraction of water and nitrates using Primene JMT, which completely eliminates these problem. Final step was calcination in air of obtained microspheres of gels to triuranium octaoxide.

Spatial and temporal dynamics of nitrate fluxes in a mesoscale catchment

NASA Astrophysics Data System (ADS)

Muller, C.; Musolff, A.; Strachauer, U.; Brauns, M.; Tarasova, L.; Merz, R.; Knoeller, K.

2017-12-01

Spatially and temporally variable and often superimposing processes like mobilization and turnover of N-species strongly affect nitrate fluxes at catchment outlets. It remains thus challenging to determine dominant nitrate sources to derive an effective river management. Here, we combine data sets from two spatially highly resolved key-date monitoring campaigns of nitrate fluxes along a mesoscale catchment in Germany with four years of monitoring data from two representative sites within the catchment. The study area is characterized by a strong land use gradient from pristine headwaters to lowland sub-catchments with intense agricultural land use and wastewater sources. Flow conditions were assessed by a hydrograph separation showing the clear dominance of base flow during both investigations. However, the absolute amounts of discharge differed significantly from each other (outlet: 1.42 m³ s-1 versus 0.43 m³ s-1). Nitrate concentration and flux in the headwater was found to be low. In contrast, nitrate loads further downstream originate from anthropogenic sources such as effluents from wastewater treatment plants (WWTP) and agricultural land use. The agricultural contribution did not vary in terms of nitrate concentration and isotopic signature between the years but in terms of flux. The contrasting amounts of discharge between the years led to a strongly increased relative wastewater contribution with decreasing discharge. This was mainly manifested in elevated δ18O-NO3- values downstream from the wastewater discharge. The four-year monitoring at two sides clearly indicates the chemostatic character of the agricultural N-source and its distinct, yet stable isotopic fingerprint. Denitrification was found to play no dominant role only for controlling nitrate loads in the river. The spatially highly resolved monitoring approach helped to accurately define hot spots of nitrate inputs into the stream while the long-term information allowed a classification of the results with respect to the seasonal N-dynamics in the catchment.

Synthesis and Characterization of High Energy Polymers.

DTIC Science & Technology

1981-03-31

have been poly(cis-l ,4-butadiene), CTBN , and HTPB I=1s of nitrated and DD I JA 7 1473 EDITION OF I NOV 45 1& OBSOLETE SE1CURITY CLAIISIFI1 TIN Of...cis- butadiene), PB, carboxy-tenninated poly(butadiene-co-acrylonitrile), CTBN and hydroxy-terminated poly(butadiene), HTPB. For the nitration of...evaluation. Progress Report I. Nitration of Diene Polymers and Cgolymers A. Nitromercuration of Carboxy-terminated Butadiene Acrylonitrile Copolymer ( CTBN

AMINE EXTRACTION OF PLUTONIUM FROM NITRIC ACID SOLUTIONS LOADING AND STRIPPING EXPERIMENTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wilson, A.S.

1961-01-19

Information is presented on a suitable amine processing system for plutonium nitrate. Experiments with concentrated plutonium nitrate solutions show that trilaurylamine (TLA) - xylene solvent systems did not form a second organic phase. Experiments are also reported with tri-noctylamine (TnOA)-xylene and TLA-Amsco - octyl alcohol. Two organic phases appear in both these systems at high plutonium nitrate concentrations. Data are tabulated from loading and stripping experiments. (J.R.D.)

Metabolism of ethylbenzene by human liver microsomes and recombinant human cytochrome P450s (CYP).

PubMed

Sams, Craig; Loizou, George D; Cocker, John; Lennard, Martin S

2004-03-07

The enzyme kinetics of the initial hydroxylation of ethylbenzene to form 1-phenylethanol were determined in human liver microsomes. The individual cytochrome P450 (CYP) forms catalysing this reaction were identified using selective inhibitors and recombinant preparations of hepatic CYPs. Production of 1-phenylethanol in hepatic microsomes exhibited biphasic kinetics with a high affinity, low Km, component (mean Km = 8 microM; V(max) = 689 pmol/min/mg protein; n = 6 livers) and a low affinity, high Km, component (Km = 391 microM; V(max) = 3039 pmol/min/mg protein; n = 6). The high-affinity component was inhibited 79%-95% (mean 86%) by diethyldithiocarbamate, and recombinant CYP2E1 was shown to metabolise ethylbenzene with low Km (35 microM), but also low (max) (7 pmol/min/pmol P450), indicating that this isoform catalysed the high-affinity component. Recombinant CYP1A2 and CYP2B6 exhibited high V(max) (88 and 71 pmol/min/pmol P450, respectively) and high Km (502 and 219 microM, respectively), suggesting their involvement in catalysing the low-affinity component. This study has demonstrated that CYP2E1 is the major enzyme responsible for high-affinity side chain hydroxylation of ethylbenzene in human liver microsomes. Activity of this enzyme in the population is highly variable due to induction or inhibition by physiological factors, chemicals in the diet or some pharmaceuticals. This variability can be incorporated into the risk assessment process to improve the setting of occupational exposure limits and guidance values for biological monitoring.

High-Speed Lateral Flow Strategy for a Fast Biosensing with an Improved Selectivity and Binding Affinity.

PubMed

Cho, Dong Guk; Yoo, Haneul; Lee, Haein; Choi, Yeol Kyo; Lee, Minju; Ahn, Dong June; Hong, Seunghun

2018-05-10

We report a high-speed lateral flow strategy for a fast biosensing with an improved selectivity and binding affinity even under harsh conditions. In this strategy, biosensors were fixed at a location away from the center of a round shape disk, and the disk was rotated to create the lateral flow of a target solution on the biosensors during the sensing measurements. Experimental results using the strategy showed high reaction speeds, high binding affinity, and low nonspecific adsorptions of target molecules to biosensors. Furthermore, binding affinity between target molecules and sensing molecules was enhanced even in harsh conditions such as low pH and low ionic strength conditions. These results show that the strategy can improve the performance of conventional biosensors by generating high-speed lateral flows on a biosensor surface. Therefore, our strategy can be utilized as a simple but powerful tool for versatile bio and medical applications.

High affinity ligands from in vitro selection: Complex targets

PubMed Central

Morris, Kevin N.; Jensen, Kirk B.; Julin, Carol M.; Weil, Michael; Gold, Larry

1998-01-01

Human red blood cell membranes were used as a model system to determine if the systematic evolution of ligands by exponential enrichment (SELEX) methodology, an in vitro protocol for isolating high-affinity oligonucleotides that bind specifically to virtually any single protein, could be used with a complex mixture of potential targets. Ligands to multiple targets were generated simultaneously during the selection process, and the binding affinities of these ligands for their targets are comparable to those found in similar experiments against pure targets. A secondary selection scheme, deconvolution-SELEX, facilitates rapid isolation of the ligands to targets of special interest within the mixture. SELEX provides high-affinity compounds for multiple targets in a mixture and might allow a means for dissecting complex biological systems. PMID:9501188

Haemoglobin Pierre-Benite--a high affinity variant associated with relative polycythaemia.

PubMed

Beard, M E; Potter, H C; Spearing, R L; Brennan, S O

2001-12-01

This is the second reported example of Hb Pierre--Benite (beta90 Glu-->Asp). This mutation is associated with increased oxygen affinity and polycythaemia. No instability was found and there was no charge shift detected by cellulose acetate electrophoresis at pH 8.3. The mutation was however, clearly indicated by electrospray ionization mass spectrometry (ESI MS), which showed an abnormal beta chain with a 14 Da decrease in mass. Blood volume studies documented a relative rather than a true polycythaemia and this finding has been reported in at least two other high affinity haemoglobin variants--Hb Heathrow and Hb Rahere. This finding led to delay in diagnosis because high oxygen affinity variants are conventionally considered to cause a true polycythaemia.

Interaction of nitrate and folate on the risk of breast cancer among postmenopausal women

PubMed Central

Inoue-Choi, Maki; Ward, Mary H.; Cerhan, James R.; Weyer, Peter J.; Anderson, Kristin E.; Robien, Kim

2012-01-01

Ingested nitrate can be endogenously reduced to nitrite, which may form N-nitroso compounds, known potent carcinogens. However, some studies have reported no or inverse associations between dietary nitrate intake and cancer risk. These associations may be confounded by a protective effect of folate, which plays a vital role in DNA repair. We evaluated the interaction of dietary and water nitrate intake with total folate intake on breast cancer risk in the Iowa Women’s Health Study. Dietary intake was assessed at study baseline. Nitrate intake from public water was assessed using a historical database on Iowa municipal water supplies. After baseline exclusions, 34,388 postmenopausal women and 2,875 incident breast cancers were included. Overall, neither dietary nor water nitrate was associated with breast cancer risk. Among those with folate intake ≥400 μg/d, breast cancer risk was significantly increased in public water users with the highest nitrate quintile (HR=1.40, 95%CI=1.05–1.87) and private well users (HR=1.38, 95%CI=1.05–1.82) compared to public water users with the lowest nitrate quintile; in contrast, there was no association among those with lower folate intake. Our findings do not support a previous report of increased risk of breast cancer among individuals with high dietary nitrate but low folate intake. PMID:22642949

Is dietary nitrate/nitrite exposure a risk factor for development of thyroid abnormality? A systematic review and meta-analysis.

PubMed

Bahadoran, Zahra; Mirmiran, Parvin; Ghasemi, Asghar; Kabir, Ali; Azizi, Fereidoun; Hadaegh, Farzad

2015-05-01

The potential effects of inorganic nitrate/nitrite on global health are a much debated issue. In addition to possible methemoglobinemia and carcinogenic properties, anti-thyroid effects of nitrate/nitrite have been suggested. Considering the growing significance of nitrate/nitrite and since there is no comprehensive review in data available, clarifying the effect of nitrate/nitrite on thyroid disorder outcomes is essential. Therefore, we conducted this systematic review of experimental and clinical studies, and a meta-analysis of relevant cohort and cross-sectional studies investigating the association of nitrate/nitrite exposure and thyroid function. Most animal studies show that high exposure (~10-600 times of acceptable daily intake) to nitrate/nitrite induces anti-thyroid effects, including decreased serum level of thyroid hormones and histomorphological changes in thyroid gland; however no similar observations have been documented in humans. Based on our meta-analysis, no significant association was observed between nitrate exposure and the risk of thyroid cancer, hyper- and hypothyroidism; findings from three cohort studies however showed a significant association between higher exposure to nitrite and the risk of thyroid cancer (risk = 1.48, 95% confidence interval = 1.09-2.02, P = 0.012). Additional research is needed to clarify the association between nitrate/nitrite exposures and both thyroid function and cancer. Copyright © 2015 Elsevier Inc. All rights reserved.

Interaction of nitrate and folate on the risk of breast cancer among postmenopausal women.

PubMed

Inoue-Choi, Maki; Ward, Mary H; Cerhan, James R; Weyer, Peter J; Anderson, Kristin E; Robien, Kim

2012-01-01

Ingested nitrate can be endogenously reduced to nitrite, which may form N-nitroso compounds, known potent carcinogens. However, some studies have reported no or inverse associations between dietary nitrate intake and cancer risk. These associations may be confounded by a protective effect of folate, which plays a vital role in DNA repair. We evaluated the interaction of dietary and water nitrate intake with total folate intake on breast cancer risk in the Iowa Women's Health Study. Dietary intake was assessed at study baseline. Nitrate intake from public water was assessed using a historical database on Iowa municipal water supplies. After baseline exclusions, 34,388 postmenopausal women and 2,875 incident breast cancers were included. Overall, neither dietary nor water nitrate was associated with breast cancer risk. Among those with folate intake ≥400 μg/day, breast cancer risk was significantly increased in public water users with the highest nitrate quintile (HR = 1.40, 95% CI = 1.05-1.87) and private well users (HR = 1.38, 95% CI = 1.05-1.82) compared to public water users with the lowest nitrate quintile; in contrast, there was no association among those with lower folate intake. Our findings do not support a previous report of increased risk of breast cancer among individuals with high dietary nitrate but low folate intake.

Simulating long term nitrate-N contamination processes in the Woodville Karst Plain using CFPv2 with UMT3D

NASA Astrophysics Data System (ADS)

Xu, Zexuan; Hu, Bill X.; Davis, Hal; Cao, Jianhua

2015-05-01

A research version of CFP (Conduit Flow Process) code, CFPv2, is applied with UMT3D to simulate long term (1966-2018) nitrate-N contamination transport processes in the Woodville Karst Plain (WKP), northern Florida, where karst conduit networks are well developed. Groundwater flow in the WKP limestone porous matrix is simulated using Darcy's law, and non-laminar flow within conduits is described by Darcy-Weisbach equation. Nitrate-N conduit transport and advective exchanges of groundwater and nitrate-N between conduits and limestone matrix are calculated by CFPv2 and UMT3D, instead of MODFLOW and MT3DMS since Reynolds numbers for flows in conduits are over the criteria of laminar flow. The developed numerical model is calibrated by field observations and then applied to simulate nitrate-N transport in the WKP. The numerical simulations verify the theories that two sprayfields near the City of Tallahassee and septic tanks in the rural area are major nitrate-N point sources within the WKP. High nitrate-N concentrations occur near Lost Creek Sink, and conduits of Wakulla Spring and Spring Creek Springs where aquifer discharge groundwater. Conduit networks control nitrate-N transport and regional contaminant distributions in the WKP, as nitrate-N is transported through conduits rapidly and spread over large areas.

Denitrification in a Sand and Gravel Aquifer

PubMed Central

Smith, Richard L.; Duff, John H.

1988-01-01

Denitrification was assayed by the acetylene blockage technique in slurried core material obtained from a freshwater sand and gravel aquifer. The aquifer, which has been contaminated with treated sewage for more than 50 years, had a contaminant plume greater than 3.5-km long. Near the contaminant source, groundwater nitrate concentrations were greater than 1 mM, whereas 0.25 km downgradient the central portion of the contaminant plume was anoxic and contained no detectable nitrate. Samples were obtained along the longitudinal axis of the plume (0 to 0.25 km) at several depths from four sites. Denitrification was evident at in situ nitrate concentrations at all sites tested; rates ranged from 2.3 to 260 pmol of N2O produced (g of wet sediment)−1 h−1. Rates were highest nearest the contaminant source and decreased with increasing distance downgradient. Denitrification was the predominant nitrate-reducing activity; no evidence was found for nitrate reduction to ammonium at any site. Denitrifying activity was carbon limited and not nitrate limited, except when the ambient nitrate level was less than the detection limit, in which case, even when amended with high concentrations of glucose and nitrate, the capacity to denitrify on a short-term basis was lacking. These results demonstrate that denitrification can occur in groundwater systems and, thereby, serve as a mechanism for nitrate removal from groundwater. PMID:16347621

The feasibility of applying immature yard-waste compost to remove nitrate from agricultural drainage effluents: A preliminary assessment

USGS Publications Warehouse

Tsui, L.; Krapac, I.G.; Roy, W.R.

2007-01-01

Nitrate is a major agricultural pollutant found in drainage waters. Immature yard-waste compost was selected as a filter media to study its feasibility for removing nitrate from drainage water. Different operation parameters were tested to examine the denitrification efficiency, including the amounts of compost packed in columns, the flow rate, and the compost storage periods. The experimental results suggested that hydraulic retention time was the major factor to determine the extent of nitrate removal, although the amount of compost packed could also contribute to the nitrate removal efficiency. The effluent nitrate concentration increased as the flow rate decreased, and the compost column reduced nitrate concentrations from 20 mg/L to less than 5 mg/L within 1.5 h. The solution pH increased at the onset of experiment because of denitrification, but stabilized at a pH of about 7.8, suggesting that the compost had a buffering capacity to maintain a suitable pH for denitrification. Storing compost under air-dried conditions may diminish the extent nitrate removed initially, but the effects were not apparent after longer applications. It appeared that immature yard-waste compost may be a suitable material to remove nitrate from tile drainage water because of its relatively large organic carbon content, high microbial activity, and buffering capacity. ?? 2006 Elsevier B.V. All rights reserved.

RNA-seq analyses reveal insights into the function of respiratory nitrate reductase of the diazotroph Herbaspirillum seropedicae.

PubMed

Bonato, Paloma; Batista, Marcelo B; Camilios-Neto, Doumit; Pankievicz, Vânia C S; Tadra-Sfeir, Michelle Z; Monteiro, Rose Adele; Pedrosa, Fabio O; Souza, Emanuel M; Chubatsu, Leda S; Wassem, Roseli; Rigo, Liu Un

2016-09-01

Herbaspirillum seropedicae is a nitrogen-fixing β-proteobacterium that associates with roots of gramineous plants. In silico analyses revealed that H. seropedicae genome has genes encoding a putative respiratory (NAR) and an assimilatory nitrate reductase (NAS). To date, little is known about nitrate metabolism in H. seropedicae, and, as this bacterium cannot respire nitrate, the function of NAR remains unknown. This study aimed to investigate the function of NAR in H. seropedicae and how it metabolizes nitrate in a low aerated-condition. RNA-seq transcriptional profiling in the presence of nitrate allowed us to pinpoint genes important for nitrate metabolism in H. seropedicae, including nitrate transporters and regulatory proteins. Additionally, both RNA-seq data and physiological characterization of a mutant in the catalytic subunit of NAR (narG mutant) showed that NAR is not required for nitrate assimilation but is required for: (i) production of high levels of nitrite, (ii) production of NO and (iii) dissipation of redox power, which in turn lead to an increase in carbon consumption. In addition, wheat plants showed an increase in shoot dry weight only when inoculated with H. seropedicae wild type, but not with the narG mutant, suggesting that NAR is important to H. seropedicae-wheat interaction. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

Nitrate flux on the Ross Ice Shelf, Antarctica and its relation to solar cosmic rays

NASA Astrophysics Data System (ADS)

Zeller, Edward J.; Dreschhoff, Gisela A. M.; Laird, Claude M.

1986-11-01

Nitrate flux has been determined in the snow sequence deposited at Windless Bight on the Ross Ice Shelf (Antarctica). The data were obtained from on-site analysis of nitrate concentrations from a glaciological pit and a firn core spanning the time interval from midwinter 1971 to January 1986. The high resolution data can be combined with precipitation records collected from adjacent areas to provide a record of nitrate flow. The resulting time series contains a signal which corresponds to the two major solar events of 1972 and 1984. The concentration and flux profiles may be useful in studies of Antarctic ozone depletion.

Specificity and Affinity Quantification of Flexible Recognition from Underlying Energy Landscape Topography

PubMed Central

Chu, Xiakun; Wang, Jin

2014-01-01

Flexibility in biomolecular recognition is essential and critical for many cellular activities. Flexible recognition often leads to moderate affinity but high specificity, in contradiction with the conventional wisdom that high affinity and high specificity are coupled. Furthermore, quantitative understanding of the role of flexibility in biomolecular recognition is still challenging. Here, we meet the challenge by quantifying the intrinsic biomolecular recognition energy landscapes with and without flexibility through the underlying density of states. We quantified the thermodynamic intrinsic specificity by the topography of the intrinsic binding energy landscape and the kinetic specificity by association rate. We found that the thermodynamic and kinetic specificity are strongly correlated. Furthermore, we found that flexibility decreases binding affinity on one hand, but increases binding specificity on the other hand, and the decreasing or increasing proportion of affinity and specificity are strongly correlated with the degree of flexibility. This shows more (less) flexibility leads to weaker (stronger) coupling between affinity and specificity. Our work provides a theoretical foundation and quantitative explanation of the previous qualitative studies on the relationship among flexibility, affinity and specificity. In addition, we found that the folding energy landscapes are more funneled with binding, indicating that binding helps folding during the recognition. Finally, we demonstrated that the whole binding-folding energy landscapes can be integrated by the rigid binding and isolated folding energy landscapes under weak flexibility. Our results provide a novel way to quantify the affinity and specificity in flexible biomolecular recognition. PMID:25144525

Specificity and affinity quantification of flexible recognition from underlying energy landscape topography.

PubMed

Chu, Xiakun; Wang, Jin

2014-08-01

Flexibility in biomolecular recognition is essential and critical for many cellular activities. Flexible recognition often leads to moderate affinity but high specificity, in contradiction with the conventional wisdom that high affinity and high specificity are coupled. Furthermore, quantitative understanding of the role of flexibility in biomolecular recognition is still challenging. Here, we meet the challenge by quantifying the intrinsic biomolecular recognition energy landscapes with and without flexibility through the underlying density of states. We quantified the thermodynamic intrinsic specificity by the topography of the intrinsic binding energy landscape and the kinetic specificity by association rate. We found that the thermodynamic and kinetic specificity are strongly correlated. Furthermore, we found that flexibility decreases binding affinity on one hand, but increases binding specificity on the other hand, and the decreasing or increasing proportion of affinity and specificity are strongly correlated with the degree of flexibility. This shows more (less) flexibility leads to weaker (stronger) coupling between affinity and specificity. Our work provides a theoretical foundation and quantitative explanation of the previous qualitative studies on the relationship among flexibility, affinity and specificity. In addition, we found that the folding energy landscapes are more funneled with binding, indicating that binding helps folding during the recognition. Finally, we demonstrated that the whole binding-folding energy landscapes can be integrated by the rigid binding and isolated folding energy landscapes under weak flexibility. Our results provide a novel way to quantify the affinity and specificity in flexible biomolecular recognition.

Guanine nucleotide regulatory protein co-purifies with the D/sub 2/-dopamine receptor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Senogles, S.E.; Caron, M.G.

1986-05-01

The D/sub 2/-dopamine receptor from bovine anterior pituitary was purified approx.1000 fold by affinity chromatography on CMOS-Sepharose. Reconstitution of the affinity-purified receptor into phospholipid vesicles revealed the presence of high and low affinity agonist sites as detected by N-n-propylnorapomorphine (NPA) competition experiments with /sup 3/H-spiperone. High affinity agonist binding could be converted to the low affinity form by guanine nucleotides, indicating the presence of an endogenous guanine nucleotide binding protein (N protein) in the affinity-purified D/sub 2/ receptor preparations. Furthermore, this preparation contained an agonist-sensitive GTPase activity which was stimulated 2-3 fold over basal by 10 ..mu..M NPA. /sup 35/S-GTP..gamma..Smore » binding to these preparations revealed a stoichiometry of 0.4-0.7 mole N protein/mole receptor, suggesting the N protein may be specifically coupled with the purified D/sub 2/-dopamine receptor and not present as a contaminant. Pertussis toxin treatment of the affinity purified receptor preparations prevented high affinity agonist binding, as well as agonist stimulation of the GTPase activity, presumably by inactivating the associated N protein. Pertussis toxin lead to the ADP-ribosylation of a protein of 39-40K on SDS-PAGE. These findings indicate that an endogenous N protein, N/sub i/ or N/sub o/, co-purifies with the D/sub 2/-dopamine receptor which may reflect a precoupling of this receptor with an N protein within the membranes.« less

Source and transport controls on the movement of nitrate to public supply wells in selected principal aquifers of the United States

NASA Astrophysics Data System (ADS)

McMahon, P. B.; BöHlke, J. K.; Kauffman, L. J.; Kipp, K. L.; Landon, M. K.; Crandall, C. A.; Burow, K. R.; Brown, C. J.

2008-04-01

In 2003-2005, systematic studies in four contrasting hydrogeologic settings were undertaken to improve understanding of source and transport controls on nitrate movement to public supply wells (PSW) in principal aquifers of the United States. Chemical, isotopic, and age tracer data show that agricultural fertilizers and urban septic leachate were the primary sources of large nitrate concentrations in PSW capture zones at Modesto, California (Central Valley aquifer system) and York, Nebraska (High Plains aquifer). Urban septic leachate and fertilizer (possibly nonfarm) were the primary sources of large nitrate concentrations in PSW capture zones at Woodbury, Connecticut (glacial aquifer system), and Tampa, Florida (Floridan aquifer system), respectively. Nitrate fluxes to the water table were larger in agricultural settings than urban settings, indicating that it would be beneficial to reduce PSW capture zone areas in agricultural regions. Mixing calculations indicate that about 50 to 85% of the nitrate in water from the PSW could be from those modern anthropogenic sources, with the remainder coming from sources in old (>50 years) recharge or sources in young recharge in undisturbed settings such as forests. Excess N2 concentrations and age tracers showed that denitrification at Modesto occurred gradually (first-order rate constant of 0.02/a) in a thick reaction zone following a ˜30-year lag time after recharge. Denitrification generally was not an important nitrate sink at Woodbury. At York and Tampa, denitrification occurred rapidly (0.5 to 6/a) in thin reaction zones in fine-grained sediments that separated the anoxic PSW producing zones from overlying oxic, high-nitrate ground water. Particle tracking showed that a major pathway by which anthropogenic nitrate reached the York and Tampa PSW was by movement through long well screens crossing multiple hydrogeologic units (York) and by movement through karst features (Tampa), processes which reduced ground water residence times in the denitrifying zones. These results illustrate how PSW vulnerability to nitrate contamination depends on complex variations and interactions between contaminant sources, reaction rates, transit times, mixing, and perturbation of ground water flow in contrasting hydrogeologic settings.

Molecular Basis of Autophagic Cell Death in Prostate Cancer

DTIC Science & Technology

2009-03-01

lipid-binding proteinwith high affinity for phosphatidic acid (PA) and cardiolipin (CL). Previously, it has been shown that PA directly interacted...lyceride), PI (phosphatidylinositol), DAG (diacylglycerol), PI4P (PtdIns(4)P), PA ( phosphatidic acid ), PI4,5P2 (PtdIns(4,5)P2), PS (phosphatidylserine...with high affinity for phosphatidic acid and cardiolipin and less affinity for various phosphoinositides. Functional genomics analysis identified 5

Efficient T-cell receptor signaling requires a high-affinity interaction between the Gads C-SH3 domain and the SLP-76 RxxK motif.

PubMed

Seet, Bruce T; Berry, Donna M; Maltzman, Jonathan S; Shabason, Jacob; Raina, Monica; Koretzky, Gary A; McGlade, C Jane; Pawson, Tony

2007-02-07

The relationship between the binding affinity and specificity of modular interaction domains is potentially important in determining biological signaling responses. In signaling from the T-cell receptor (TCR), the Gads C-terminal SH3 domain binds a core RxxK sequence motif in the SLP-76 scaffold. We show that residues surrounding this motif are largely optimized for binding the Gads C-SH3 domain resulting in a high-affinity interaction (K(D)=8-20 nM) that is essential for efficient TCR signaling in Jurkat T cells, since Gads-mediated signaling declines with decreasing affinity. Furthermore, the SLP-76 RxxK motif has evolved a very high specificity for the Gads C-SH3 domain. However, TCR signaling in Jurkat cells is tolerant of potential SLP-76 crossreactivity, provided that very high-affinity binding to the Gads C-SH3 domain is maintained. These data provide a quantitative argument that the affinity of the Gads C-SH3 domain for SLP-76 is physiologically important and suggest that the integrity of TCR signaling in vivo is sustained both by strong selection of SLP-76 for the Gads C-SH3 domain and by a capacity to buffer intrinsic crossreactivity.

Mitochondrial respiratory pathways modulate nitrate sensing and nitrogen-dependent regulation of plant architecture in Nicotiana sylvestris

PubMed Central

Pellny, Till K; Van Aken, Olivier; Dutilleul, Christelle; Wolff, Tonja; Groten, Karin; Bor, Melike; De Paepe, Rosine; Reyss, Agnès; Van Breusegem, Frank; Noctor, Graham; Foyer, Christine H

2008-01-01

Mitochondrial electron transport pathways exert effects on carbon–nitrogen (C/N) relationships. To examine whether mitochondria–N interactions also influence plant growth and development, we explored the responses of roots and shoots to external N supply in wild-type (WT) Nicotiana sylvestris and the cytoplasmic male sterile II (CMSII) mutant, which has a N-rich phenotype. Root architecture in N. sylvestris seedlings showed classic responses to nitrate and sucrose availability. In contrast, CMSII showed an altered ‘nitrate-sensing’ phenotype with decreased sensitivity to C and N metabolites. The WT growth phenotype was restored in CMSII seedling roots by high nitrate plus sugars and in shoots by gibberellic acid (GA). Genome-wide cDNA-amplified fragment length polymorphism (AFLP) analysis of leaves from mature plants revealed that only a small subset of transcripts was altered in CMSII. Tissue abscisic acid content was similar in CMSII and WT roots and shoots, and growth responses to zeatin were comparable. However, the abundance of key transcripts associated with GA synthesis was modified both by the availability of N and by the CMSII mutation. The CMSII mutant maintained a much higher shoot/root ratio at low N than WT, whereas no difference was observed at high N. Shoot/root ratios were strikingly correlated with root amines/nitrate ratios, values of <1 being characteristic of high N status. We propose a model in which the amine/nitrate ratio interacts with GA signalling and respiratory pathways to regulate the partitioning of biomass between shoots and roots. PMID:18318685

Nitrate-containing beetroot juice reduces oxygen consumption during submaximal exercise in low but not high aerobically fit male runners

PubMed Central

Carriker, Colin R.; Vaughan, Roger A.; VanDusseldorp, Trisha A.; Johnson, Kelly E.; Beltz, Nicholas M.; McCormick, James J.; Cole, Nathan H.; Gibson, Ann L.

2016-01-01

[Purpose] To examine the effect of a 4-day NO3- loading protocol on the submaximal oxygen cost of both low fit and high fit participants at five different exercise intensities. [Methods] Eleven (6 high fit, VO2max 60.1 ± 4.6ml/kg/min; 5 low fit, VO2max 42.4 ± 3.2ml/ kg/min) participants were initially assigned to a placebo (PL; negligible NO3-) or inorganic nitrate-rich (NR; 6.2 mmol nitrate/day) group using a double-blind, placebo-controlled, crossover design. Participants completed three trials (T1, T2 and T3). T1 included a maximal aerobic capacity (VO2max) treadmill test. A 6-day washout, minimizing nitrate consumption, preceded T2. Each of the four days prior to T2 and T3, participants consumed either PL or NR with the final dose 2.5 hours prior to exercise. A 14-day washout followed T2. T2 and T3 consisted of 5-minute submaximal treadmill bouts (45, 60, 70, 80 and 85% VO2max) determined during T1. [Results] Low fit nitrate-supplemented participants consumed less oxygen (p<0.05) at lower workloads (45% and 60% VO2max) compared to placebo trials; changes were not observed in high fit participants. The two lowest intensity workloads of 45 and 60% VO2max revealed the greatest correlation (r=0.54, p=0.09 and r=0.79, p<0.05; respectively) between VO2max and change in oxygen consumption. No differences were found between conditions for heart rate, respiratory exchange ratio or rating of perceived exertion for either fitness group. [Conclusion] Nitrate consumption promotes reduced oxygen consumption at lower exercise intensities in low fit, but not high fit males. Lesser fit individuals may receive greater benefit than higher fit participants exercising at intensities <60% VO2max. PMID:28150476

Radioiodinated nondegradable gonadotropin-releasing hormone analogs: new probes for the investigation of pituitary gonadotropin-releasing hormone receptors.

PubMed

Clayton, R N; Shakespear, R A; Duncan, J A; Marshall, J C; Munson, P J; Rodbard, D

1979-12-01

Studies of pituitary plasma membrane gonadotropin-releasing hormone (GnRH) receptors using [125I]-iodo-GnRH suffer major disadvantages. Only a small (less than 25%) proportion of specific tracer binding is to high affinity sites, with more than 70% bound to low affinity sites (Ka = 1 x 10(6) M-1). [125I]Iodo-GnRH is also inactivated during incubation with pituitary plasma membrane preparations. Two superactive analongs of GnRH, substituted in positions 6 and 10, were used as the labeled ligand to overcome these problems. Both analogs bound to the same high affinity sites as GnRH on bovine pituitary plasma membranes, though the affinity of the analogs was higher than that of the natural decapeptide (Ka = 2.0 x 10(9), 6.0 x 10(9), and 3.0 x 10(8) M-1 for [D-Ser(TBu)6]des-Gly10-GnRH ethylamide, [D-Ala6]des-Gly10-GnRH ethylamide, and GnRH, respectively. The labeled analogs bound to a single class of high affinity sites with less than 15% of the specific binding being to low affinity sites (Ka approximately equal to 1 x 10(6) M-1). The labeled analogs were not inactivated during incubation with the pituitary membrane preparations. Using the analogs as tracer, a single class of high affinity sites (K1 = 4.0 x 10(9) M-1) was also demonstrated on crude 10,800 x g rat pituitary membrane preparations. Use of these analogs as both the labeled and unlabeled ligand offers substantial advantages over GnRH for investigation of GnRH receptors, allowing accurate determination of changes in their numbers and affinities under various physiological conditions.

Hb Potomac (101 Glu replaced by Asp): speculations on placental oxygen transport in carriers of high-affinity hemoglobins.

PubMed

Charache, S; Jacobson, R; Brimhall, B; Murphy, E A; Hathaway, P; Winslow, R; Jones, R; Rath, C; Simkovich, J

1978-02-01

Blood from a woman with unexplained erythrocytosis had increased oxygen affinity, but no abnormality could be detected by electrophoresis or chromatography of her hemolysate. Separation of the tryptic peptides of her beta chains disclosed two half-sized peaks in the regions of beta T-11. The faster of these was abnormal, with the structure beta 101 Glu replaced by Asp. The new hemoglobin was called "Potomac." Three of the proband's four surviving siblings and both of her children were carriers. Differences in the ratio of carrier: normal children born to male of female carriers of 23 other high-affinity hemoglobins were not significant. The high proportion of carriers in this kindred was probably due to chance alone, and not because high maternal oxygen affinity interfered with oxygen transport to fetuses with normal hemoglobin.

Biomarker Candidates of Chlamydophila pneumoniae Proteins and Protein Fragments Identified by Affinity-Proteomics Using FTICR-MS and LC-MS/MS

NASA Astrophysics Data System (ADS)

Susnea, Iuliana; Bunk, Sebastian; Wendel, Albrecht; Hermann, Corinna; Przybylski, Michael

2011-04-01

We report here an affinity-proteomics approach that combines 2D-gel electrophoresis and immunoblotting with high performance mass spectrometry to the identification of both full length protein antigens and antigenic fragments of Chlamydophila pneumoniae (C. pneumoniae). The present affinity-mass spectrometry approach effectively utilized high resolution FTICR mass spectrometry and LC-tandem-MS for protein identification, and enabled the identification of several new highly antigenic C. pneumoniae proteins that were not hitherto reported or previously detected only in other Chlamydia species, such as Chlamydia trachomatis. Moreover, high resolution affinity-MS provided the identification of several neo-antigenic protein fragments containing N- and C-terminal, and central domains such as fragments of the membrane protein Pmp21 and the secreted chlamydial proteasome-like factor (Cpaf), representing specific biomarker candidates.

Lesion-induced plasticity of high affinity choline uptake in the developing rat fascia dentata.

PubMed

Nadler, J V; Shelton, D L; Cotman, C W

1979-03-23

After removal of the perforant path input to the rat fascia dentata at the age of 11 days, cholinergic septohippocampal fibers invade the denervated area. We have examined the effect of this lesion on hemicholinium-sensitive, high affinity choline uptake and its coupling to acetylcholine synthesis, specific properties of the septohippocampal input. Removal of the ipsilateral perforant path fibers increased the velocity of high affinity choline uptake by dentate particulate preparations, usually within 1 day. Studies conducted 5--104 days after operation showed a consistent 50--65% elevation in the molecular (denervated) layer. In contrast, the choline uptake rate in the granular layer eventually decreased slightly. Calculation of choline uptake rates independently of protein (per whole region) revealed that fasciae dentatae from operated and control sides accumulated choline at approximately equal rates, but on the operated side a greater percentage was transported by structures from the molecular layer and a lesser percentage by those from the granular layer. The rate of acetylcholine synthesis from exogenous choline increased to the same extent as high affinity choline uptake from 3 days after operation onwards. The changes in high affinity choline uptake and acetylcholine synthesis coincided spatially and temporally with the reactive growth of septohippocampal fibers. Our results support the view that a perforant path lesion during development permanently alters the distribution of functional septohippocampal boutons in the fascia dentata. Acetylcholine synthesis is regulated to the same extent by high affinity choline uptake in the anomalous boutons as in normally located boutons.

A putative role for the plasma membrane potential in the control of the expression of the gene encoding the tomato high-affinity potassium transporter HAK5.

PubMed

Nieves-Cordones, Manuel; Miller, Anthony J; Alemán, Fernando; Martínez, Vicente; Rubio, Francisco

2008-12-01

A chimeric CaHAK1-LeHAK5 transporter with only 15 amino acids of CaHAK1 in the N-terminus mediates high-affinity K(+) uptake in yeast cells. Kinetic and expression analyses strongly suggest that LeHAK5 mediates a significant proportion of the high-affinity K(+) uptake shown by K(+)-starved tomato (Solanum lycopersicum) plants. The development of high-affinity K(+) uptake, putatively mediated by LeHAK5, was correlated with increased LeHAK5 mRNA levels and a more negative electrical potential difference across the plasma membrane of root epidermal and cortical cells. However, this increase in high-affinity K(+) uptake was not correlated with the root K(+) content. Thus, (i) growth conditions that result in a hyperpolarized root plasma membrane potential, such as K(+) starvation or growth in the presence of NH(4) (+), but which do not decrease the K(+) content, lead to increased LeHAK5 expression; (ii) the presence of NaCl in the growth solution, which prevents the hyperpolarization induced by K(+) starvation, also prevents LeHAK5 expression. Moreover, once the gene is induced, depolarization of the plasma membrane potential then produces a decrease in the LeHAK5 mRNA. On the basis of these results, we propose that the plant membrane electrical potential plays a role in the regulation of the expression of this gene encoding a high-affinity K(+) transporter.

Evaluation of the Origin and Fate of Nitrate in the Aquifer System of Southern Baldwin County, Alabama Using Multi-isotopic Data

NASA Astrophysics Data System (ADS)

Murgulet, D.; Tick, G. R.

2008-12-01

Continued and extensive residential and agricultural development of near-shore areas in southern Baldwin County, Alabama has led to increased inputs of nitrogen (N) to groundwater and to the Gulf of Mexico. Nitrate (NO3-) concentrations in several groundwater wells exceeded the regulatory drinking water standards (10 mg/L nitrate-N). Groundwater and surface water samples were analyzed for nitrate, phosphate, salinity, chloride, and total dissolved solids concentrations to assess the extent of nitrate contamination. Nitrogen and oxygen isotopes of nitrate (e.g., δ15N and δ18O of nitrate) were used in conjunction with other isotopic data (e.g., 13C, and 14C, and hydrogen and oxygen isotopes of groundwater) and hydrogeochemical data to place constraints on potential sources of nitrate. The δ15N and δ18O of groundwater nitrate values ranged between +3.1 and +9.6‰ and +4.2 and +8.7‰, respectively. This range of values suggests that nitrate is primarily derived from nitrification of reduced N compounds (primarily ammonia) from fertilizer and manure or septic waste. However, an overwhelming number of samples show isotopic signatures which indicate that the predominant source of nitrate in these aguifers is the fertilizer and to some extent, for deeper wells with older groundwater, the atmospheric nitrate. The narrow range of δ18O values further confirms the primary nitrate sources. The δ15N and δ18O of nitrate data indicate that denitrification was not an important processes in these aquifers. This conclusion is also supported by the 114C data which revealed relatively young groundwaters with sufficiently high oxygen levels. In the absence of denitrification and the presence of a permanent source, it is expected that the elevated groundwater nitrate concentrations will not be readily attenuated posing a potential contamination and degradation problem of coastal discharge zones into the future. The δ13C and δ18O of groundwater data indicates that water in the aquifer system of the study area is most likely to have originated from precipitation and soil infiltration through relatively localized recharge.

Using Multi-Isotope Tracer Methods to Understand the Sources of Nitrate in Aerosols, Fog and River Water in Podocarpus National Forest, Ecuador

NASA Astrophysics Data System (ADS)

Brothers, L. A.; Dominguez, G.; Fabian, P.; Thiemens, M. H.

2008-12-01

The eastern slopes of the Andean rainforests of Ecuador possess some of the highest plant biodiversity found on the planet; however, these ecosystems are in jeopardy because region is experiences one of the highest deforestation rates in South America. This rainforest characterized by high acidity and low nutrient soils and experiences natural process which are both destabilizing and stabilizing to biodiversity rendering this a unique, though sensitive environment. There is increased concern that anthropogenic activities especially biomass burning are affecting the rainforests and could lead to higher extinction rates, changes in the biodiversity and far reaching effects on the global troposphere. Measurements of nitrate and sulfate in rain and fog water have shown periods of elevated concentrations in the Podocarpus National Park near Loja, Ecuador. These high episodes contribute to annual deposition rates that are comparable to polluted regions of North America and Europe. Significant anthropogenic sources such as large scale industry or a major city, near this forest are lacking. It is believed that the majority of the nitrate and sulfate pollution is due to the large amount of biomass burning during the dry season in the Amazon Basin. In recent years it has been shown that large amount of dust is transported across the Atlantic from Africa which reaches South America. Concentration measurements do not elucidate the source of high nitrate and sulfate pollution; however, by measuring all three stable isotopes of oxygen in nitrate and sulfate from fog and river water provides a new way to examine the impacts of biomass burning on the region. By using stable isotope techniques atmospheric nitrate and sulfate can be resolved from terrestrial sources. This provides a unique way to trace the contributions from the biomass burning and farming sources. Current research at the field station, Estación Científica San Francisco in the Podocarpus National Forest monitors sulfate and nitrate concentrations in rain and fog water by standard methods to investigate water and nutrient pathways along with data from satellite and ground based remote sensing, observations and numerical models. We hope to pair this with a multi-isotope tracer method and NOAA Hysplit Back trajectories, and satellite imagery for information about the number of fires burning in the region to help identify sources of the high nitrate deposition.

Photocatalytic reduction of nitrate using titanium dioxide for regeneration of ion exchange brine.

PubMed

Yang, Ting; Doudrick, Kyle; Westerhoff, Paul

2013-03-01

Nitrate is often removed from groundwater by ion exchange (IX) before its use as drinking water. Accumulation of nitrate in IX brine reduces the efficiency of IX regeneration and the useful life of the regeneration brine. For the first time, we present a strategy to photocatalytically reduce nitrate in IX brine, thereby extending the use of the brine. Titanium dioxide (Evonik P90), acting as photocatalyst, reduced nitrate effectively in both synthetic brines and sulfate-removed IX brine when formic acid (FA) was used as the hole scavenger (i.e., electron donor) and the initial FA to nitrate molar ratio (IFNR) was 5.6. Increasing the NaCl level in the synthetic brine slowed the nitrate reduction rate without affecting by-product selectivity of ammonium and gaseous N species (e.g., N(2), N(2)O). In a non-modified IX brine, nitrate removal was greatly inhibited owing to the presence of sulfate, which competed with nitrate for active surface sites on P90 and induced aggregation of P90 nanoparticles. After removing sulfate through barium sulfate precipitation, nitrate was effectively reduced; approximately 3.6 × 10(24) photons were required to reduce each mole of nitrate to 83% N Gases and 17% NH(4)(+). To make optimum use of FA and control the residual FA level in treated brine, the IFNR was varied. High IFNRs (e.g., 4, 5.6) were found to be more efficient for nitrate reduction but left higher residual FA in brine. IX column tests were performed to investigate the impact of residual FA for brine reuse. The residual FA in the brine did not significantly affect the nitrate removal capacity of IX resins, and formate contamination of treated water could be eliminated by rinsing with one bed volume of fresh brine. Copyright © 2012 Elsevier Ltd. All rights reserved.

Effects of dietary nitrate supplementation on symptoms of acute mountain sickness and basic physiological responses in a group of male adolescents during ascent to Mount Everest Base Camp.

PubMed

Hennis, Philip J; Mitchell, Kay; Gilbert-Kawai, Edward; Bountziouka, Vassiliki; Wade, Angie; Feelisch, Martin; Grocott, Michael P; Martin, Daniel S

2016-11-30

The purpose of this study was to investigate the effects of dietary nitrate supplementation, in the form of beetroot juice, on acute mountain sickness (AMS) symptoms and physiological responses, in a group of young males trekking to Mount Everest Base Camp (EBC). Forty healthy male students (mean age (SD): 16 (1) yrs) trekked to EBC over 11 days. Following an overnight fast, each morning participants completed the Lake Louise AMS questionnaire and underwent a series of physiological tests: resting blood pressure as well as resting and exercising heart rate, respiratory rate, and peripheral oxygen saturation. The exercise test consisted of a standardised 2-min stepping protocol and measurements were taken in the last 10 s. Participants in the intervention arm of the study consumed 140 ml of concentrated beetroot juice daily, containing approximately 10 mmol of nitrate, while those in the control arm consumed 140 ml of concentrated blackcurrant cordial with negligible nitrate content. Drinks were taken for the first seven days at high altitude (days 2-8), in two equal doses; one with breakfast, and one with the evening meal. Mixed modelling revealed no significant between-groups difference in the incidence of AMS (Odds Ratio - nitrate vs. 1.16 (95% CI: 0.59; 2.29)). Physiological changes occurring during ascent to high altitude generally were not significantly different between the two groups (Model Coef (95% CI) - average difference nitrate vs. systolic blood pressure, 0.16 (-4.47; 4.79); peripheral oxygen saturation, 0.28 (-0.85; 1.41); heart rate, -0.48 (-8.47; 7.50) (Model Coef (95% CI) - relative difference nitrate vs. ventilatory rate, 0.95 (0.82; 1.08)). Modelling revealed that diastolic blood pressure was 3.37 mmHg (0.24; 6.49) higher for participants in the beetroot juice, however this difference was no larger than that found at baseline and no interaction effect was observed. Supplementation with dietary nitrate did not significantly change symptoms of AMS or alter key physiological variables, in a group of adolescent males during a high altitude trekking expedition. There was no evidence of harm from dietary nitrate supplementation in this context. Given the wide confidence intervals in all models, a larger sample size would be required to exclude a false negative result. Our data suggest that prolonged oral nitrate supplementation is safe and feasible at altitude but has little physiological or clinical effect. Copyright © 2016 Elsevier Inc. All rights reserved.

Application of anaerobic digested residues on safe food production.

PubMed

Shi, Ya-juan; Lu, Yong-long; Liang, Dan

2002-01-01

Experiments were conducted in pot culture and field plots to study the effects of Anaerobic Digested Residues (ADR) on nitrate accumulation in leaf vegetables, which is critical for the safety of food. The results showed that compared to chemical fertilizer, ADR could decrease the nitrate accumulation in rape and spinach. Furthermore, nitrate content in plant tissue was increased with the increase of percentage of chemical nitrogen in the mixture of chemical fertilizer and ADR. A comparison of spraying digested slurry with irrigation showed that spraying method could reduce the nitrate content of rape, however, a reverse result was found in spinach. The nitrate accumulation in rape affected by ADR was more apparent in high fertility soil than that in low fertility one. To regulate the nitrate accumulation in plant, it was more apparent in rape under greenhouse cultivation, while more apparent in spinach under open-air cultivation. The results demonstrated that the ADR was effective in the safe food production and it may convert the technology to be more profitable.

Induction of the Nitrate Assimilation nirA Operon and Protein-Protein Interactions in the Maturation of Nitrate and Nitrite Reductases in the Cyanobacterium Anabaena sp. Strain PCC 7120.

PubMed

Frías, José E; Flores, Enrique

2015-07-01

Nitrate is widely used as a nitrogen source by cyanobacteria, in which the nitrate assimilation structural genes frequently constitute the so-called nirA operon. This operon contains the genes encoding nitrite reductase (nirA), a nitrate/nitrite transporter (frequently an ABC-type transporter; nrtABCD), and nitrate reductase (narB). In the model filamentous cyanobacterium Anabaena sp. strain PCC 7120, which can fix N2 in specialized cells termed heterocysts, the nirA operon is expressed at high levels only in media containing nitrate or nitrite and lacking ammonium, a preferred nitrogen source. Here we examined the genes downstream of the nirA operon in Anabaena and found that a small open reading frame of unknown function, alr0613, can be cotranscribed with the operon. The next gene in the genome, alr0614 (narM), showed an expression pattern similar to that of the nirA operon, implying correlated expression of narM and the operon. A mutant of narM with an insertion mutation failed to produce nitrate reductase activity, consistent with the idea that NarM is required for the maturation of NarB. Both narM and narB mutants were impaired in the nitrate-dependent induction of the nirA operon, suggesting that nitrite is an inducer of the operon in Anabaena. It has previously been shown that the nitrite reductase protein NirA requires NirB, a protein likely involved in protein-protein interactions, to attain maximum activity. Bacterial two-hybrid analysis confirmed possible NirA-NirB and NarB-NarM interactions, suggesting that the development of both nitrite reductase and nitrate reductase activities in cyanobacteria involves physical interaction of the corresponding enzymes with their cognate partners, NirB and NarM, respectively. Nitrate is an important source of nitrogen for many microorganisms that is utilized through the nitrate assimilation system, which includes nitrate/nitrite membrane transporters and the nitrate and nitrite reductases. Many cyanobacteria assimilate nitrate, but regulation of the nitrate assimilation system varies in different cyanobacterial groups. In the N2-fixing, heterocyst-forming cyanobacteria, the nirA operon, which includes the structural genes for the nitrate assimilation system, is expressed in the presence of nitrate or nitrite if ammonium is not available to the cells. Here we studied the genes required for production of an active nitrate reductase, providing information on the nitrate-dependent induction of the operon, and found evidence for possible protein-protein interactions in the maturation of nitrate reductase and nitrite reductase. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Analysis of Protein Interactions with Picomolar Binding Affinity by Fluorescence-Detected Sedimentation Velocity

PubMed Central

2014-01-01

The study of high-affinity protein interactions with equilibrium dissociation constants (KD) in the picomolar range is of significant interest in many fields, but the characterization of stoichiometry and free energy of such high-affinity binding can be far from trivial. Analytical ultracentrifugation has long been considered a gold standard in the study of protein interactions but is typically applied to systems with micromolar KD. Here we present a new approach for the study of high-affinity interactions using fluorescence detected sedimentation velocity analytical ultracentrifugation (FDS-SV). Taking full advantage of the large data sets in FDS-SV by direct boundary modeling with sedimentation coefficient distributions c(s), we demonstrate detection and hydrodynamic resolution of protein complexes at low picomolar concentrations. We show how this permits the characterization of the antibody–antigen interactions with low picomolar binding constants, 2 orders of magnitude lower than previously achieved. The strongly size-dependent separation and quantitation by concentration, size, and shape of free and complex species in free solution by FDS-SV has significant potential for studying high-affinity multistep and multicomponent protein assemblies. PMID:24552356

ECL-IAA and ECL-GADA Can Identify High-Risk Single Autoantibody-Positive Relatives in the TrialNet Pathway to Prevention Study.

PubMed

Steck, Andrea K; Fouts, Alexandra; Miao, Dongmei; Zhao, Zhiyuan; Dong, Fran; Sosenko, Jay; Gottlieb, Peter; Rewers, Marian J; Yu, Liping

2016-07-01

Relatives with single positive islet autoantibodies have a much lower risk of progression to diabetes than those with multiple autoantibodies. TrialNet subjects positive for single autoantibody to insulin (mIAA) (n = 50) or single autoantibody to glutamic acid decarboxylase (GADA) (n = 50) were analyzed using new electrochemiluminescence (ECL) assays (ECL-IAA and ECL-GADA, respectively) at their initial visit and longitudinally over time. Affinity assays were performed on a subset of single autoantibody-positive subjects at initial and most recent visits. After a mean follow-up of 5.3 years, 20 subjects developed type 1 diabetes. Among either single GADA or single mIAA subjects, those who were positive in the ECL assay showed higher affinity at the initial visit, and affinity results stayed consistent over time. No converting events from low to high or high to low affinity were seen over time. Confirmed positivity for ECL is associated with high affinity and can help staging of risk for type 1 diabetes in single autoantibody-positive subjects.

High-affinity RNA aptamers to C-reactive protein (CRP): newly developed pre-elution methods for aptamer selection

NASA Astrophysics Data System (ADS)

Orito, N.; Umekage, S.; Sato, K.; Kawauchi, S.; Tanaka, H.; Sakai, E.; Tanaka, T.; Kikuchi, Y.

2012-03-01

We have developed a modified SELEX (systematic evolution of ligands by exponential enrichment) method to obtain RNA aptamers with high affinity to C-reactive protein (CRP). CRP is a clinical biomarker present in plasma, the level of which increases in response to infections and noninfectious inflammation. The CRP level is also an important prognostic indicator in patients with several syndromes. At present, CRP content in blood is measured immunochemically using antibodies. To develop a more sensitive method using RNA aptamers, we have attempted to obtain high-affinity RNA aptamers to CRP. We succeeded in obtaining an RNA aptamer with high affinity to CRP using a CRP-immobilized Sepharose column and pre-elution procedure. Pre-elution is a method that removes the weak binding portion from a selected RNA population by washing for a short time with buffer containing CRP. By surface plasmon-resonance (SPR) analysis, the affinity constant of this aptamer for CRP was calculated to be KD = 2.25×10-9 (M). The secondary structure, contact sites with CRP protein, and application of this aptamer will be described.

High affinity IgM(+) memory B cells are generated through a germinal center-dependent pathway.

PubMed

Hara, Yasushi; Tashiro, Yasuyuki; Murakami, Akikazu; Nishimura, Miyuki; Shimizu, Takeyuki; Kubo, Masato; Burrows, Peter D; Azuma, Takachika

2015-12-01

During a T cell-dependent immune response, B cells undergo clonal expansion and selection and the induction of isotype switching and somatic hypermutation (SHM). Although somatically mutated IgM(+) memory B cells have been reported, it has not been established whether they are really high affinity B cells. We tracked (4-hydroxy-3-nitrophenyl) acetyl hapten-specific GC B cells from normal immunized mice based on affinity of their B cell receptor (BCR) and performed BCR sequence analysis. SHM was evident by day 7 postimmunization and increased with time, such that high affinity IgM(+) as well as IgG(+) memory B cells continued to be generated up to day 42. In contrast, class-switch recombination (CSR) was almost completed by day 7 and then the ratio of IgG1(+)/IgM(+) GC B cells remained unchanged. Together these findings suggest that IgM(+) B cells undergo SHM in the GC to generate high affinity IgM(+) memory cells and that this process continues even after CSR is accomplished. Copyright © 2015 Elsevier Ltd. All rights reserved.

In situ mobility of uranium in the presence of nitrate following sulfate-reducing conditions.

PubMed

Paradis, Charles J; Jagadamma, Sindhu; Watson, David B; McKay, Larry D; Hazen, Terry C; Park, Melora; Istok, Jonathan D

2016-04-01

Reoxidation and mobilization of previously reduced and immobilized uranium by dissolved-phase oxidants poses a significant challenge for remediating uranium-contaminated groundwater. Preferential oxidation of reduced sulfur-bearing species, as opposed to reduced uranium-bearing species, has been demonstrated to limit the mobility of uranium at the laboratory scale yet field-scale investigations are lacking. In this study, the mobility of uranium in the presence of nitrate oxidant was investigated in a shallow groundwater system after establishing conditions conducive to uranium reduction and the formation of reduced sulfur-bearing species. A series of three injections of groundwater (200 L) containing U(VI) (5 μM) and amended with ethanol (40 mM) and sulfate (20 mM) were conducted in ten test wells in order to stimulate microbial-mediated reduction of uranium and the formation of reduced sulfur-bearing species. Simultaneous push-pull tests were then conducted in triplicate well clusters to investigate the mobility of U(VI) under three conditions: 1) high nitrate (120 mM), 2) high nitrate (120 mM) with ethanol (30 mM), and 3) low nitrate (2 mM) with ethanol (30 mM). Dilution-adjusted breakthrough curves of ethanol, nitrate, nitrite, sulfate, and U(VI) suggested that nitrate reduction was predominantly coupled to the oxidation of reduced-sulfur bearing species, as opposed to the reoxidation of U(IV), under all three conditions for the duration of the 36-day tests. The amount of sulfate, but not U(VI), recovered during the push-pull tests was substantially more than injected, relative to bromide tracer, under all three conditions and further suggested that reduced sulfur-bearing species were preferentially oxidized under nitrate-reducing conditions. However, some reoxidation of U(IV) was observed under nitrate-reducing conditions and in the absence of detectable nitrate and/or nitrite. This suggested that reduced sulfur-bearing species may not be fully effective at limiting the mobility of uranium in the presence of dissolved and/or solid-phase oxidants. The results of this field study confirmed those of previous laboratory studies which suggested that reoxidation of uranium under nitrate-reducing conditions can be substantially limited by preferential oxidation of reduced sulfur-bearing species. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

In situ mobility of uranium in the presence of nitrate following sulfate-reducing conditions

DOE PAGES

Paradis, Charles J.; Jagadamma, Sindhu; Watson, David B.; ...

2016-02-11

Reoxidation and mobilization of previously reduced and immobilized uranium by dissolved phase oxidants poses a significant challenge for remediating uranium-contaminated groundwater. Preferential oxidation of reduced sulfur-bearing species, as opposed to reduced uranium bearing species, has been demonstrated to limit the mobility of uranium at the laboratory scale yet field-scale investigations are lacking. Here in this study, the mobility of uranium in the presence of nitrate oxidant was investigated in a shallow groundwater system after establishing conditions conducive to uranium reduction and the formation of reduced sulfur-bearing species. A series of three injections of groundwater (200 L) containing U(VI) (5 μM)more » and amended with ethanol (40 mM) and sulfate (20 mM) were conducted in ten test wells in order to stimulate microbial mediated reduction of uranium and the formation of reduced sulfur-bearing species. Simultaneous push-pull tests were then conducted in triplicate well clusters to investigate the mobility of U(VI) under three conditions: 1) high nitrate (120 mM), 2) high nitrate (120 mM) with ethanol (30 mM), and 3) low nitrate (2 mM) with ethanol (30 mM). Dilution-adjusted breakthrough curves of ethanol, nitrate, nitrite, sulfate, and U(VI) suggested that nitrate reduction was predominantly coupled to the oxidation of reduced-sulfur bearing species, as opposed to the reoxidation of U(IV), under all three conditions for the duration of the 36-day tests. The amount of sulfate, but not U(VI), recovered during the push-pull tests was substantially more than injected, relative to bromide tracer, under all three conditions and further suggested that reduced sulfur-bearing species were preferentially oxidized under nitrate-reducing conditions. However, some reoxidation of U(IV) was observed under nitrate-reducing conditions and in the absence of detectable nitrate and/or nitrite. This suggested that reduced sulfur-bearing species may not be fully effective at limiting the mobility of uranium in the presence of dissolved and/or solid-phase oxidants. Lastly, the results of this field study confirmed those of previous laboratory studies which suggested that reoxidation of uranium under nitrate-reducing conditions can be substantially limited by preferential oxidation of reduced sulfur-bearing species.« less

Seasonal dynamics of nitrate and ammonium ion concentrations in soil solutions collected using MacroRhizon suction cups.

PubMed

Kabala, Cezary; Karczewska, Anna; Gałka, Bernard; Cuske, Mateusz; Sowiński, Józef

2017-07-01

The aims of the study were to analyse the concentration of nitrate and ammonium ions in soil solutions obtained using MacroRhizon miniaturized composite suction cups under field conditions and to determine potential nitrogen leaching from soil fertilized with three types of fertilizers (standard urea, slow-release urea, and ammonium nitrate) at the doses of 90 and 180 kg ha -1 , applied once or divided into two rates. During a 3-year growing experiment with sugar sorghum, the concentration of nitrate and ammonium ions in soil solutions was the highest with standard urea fertilization and the lowest in variants fertilized with slow-release urea for most of the months of the growing season. Higher concentrations of both nitrogen forms were noted at the fertilizer dose of 180 kg ha -1 . One-time fertilization, at both doses, resulted in higher nitrate concentrations in June and July, while dividing the dose into two rates resulted in higher nitrate concentrations between August and November. The highest potential for nitrate leaching during the growing season was in July. The tests confirmed that the miniaturized suction cups MacroRhizon are highly useful for routine monitoring the concentration of nitrate and ammonium ions in soil solutions under field conditions.

Nitrates in drinking water and risk of death from rectal cancer in Taiwan.

PubMed

Kuo, Hsin-Wei; Wu, Trong-Neng; Yang, Chun-Yuh

2007-10-01

The relationship between nitrate levels in drinking water and rectal cancer development has been inconclusive. A matched case-control and nitrate ecology study was used to investigate the association between mortality attributed to rectal cancer and drinking-water nitrate exposure in Taiwan. All deaths due to rectal cancer of Taiwan residents from 1999 through 2003 were obtained from the Bureau of Vital Statistics of the Taiwan Provincial Department of Health. Controls were deaths from other causes and were pair matched to the cancer cases by gender, year of birth, and year of death. Each matched control was selected randomly from the set of possible controls for each case. Data on nitrate-nitrogen (NO3-N) levels in drinking water throughout Taiwan were collected from Taiwan Water Supply Corporation (TWSC). The municipality of residence for cancer cases and controls was assumed to be the source of the subject's nitrate exposure via drinking water. The adjusted odds ratios for rectal cancer death for those with high nitrate levels in their drinking water, as compared to the lowest tertile, were 1.22 (0.98-1.52) and 1.36 (1.08-1.70), respectively. The findings of this study warrant further investigation of the role of nitrates in drinking water in the etiology of rectal cancer in Taiwan.

Diminished Stream Nitrate Concentrations Linked to Dissolved Organic Carbon Dynamics After Leaf Fall

NASA Astrophysics Data System (ADS)

Sebestyen, S. D.; Shanley, J. B.; Boyer, E. W.; Doctor, D. H.; Kendall, C.

2004-05-01

Thermodynamic coupling of the nitrogen and carbon cycles has broad implications for controls on catchment nutrient fluxes. In the northeast US, leaf fall occurs in early October and the availability of organic carbon increases as the leaves decompose. At the Sleepers River Research Watershed in northeastern Vermont (USA), we sampled stream chemistry from seven nested catchments to determine how stream dissolved organic carbon (DOC) and nitrate vary as a function of flow conditions, land-use, and basin size in response to leaf fall. Following leaf fall, nitrate concentration patterns were quantitatively different from other times of the year. Under baseflow conditions, stream and soil water DOC concentrations were higher than normal, whereas nitrate concentrations declined sharply at the five smallest catchments and more modestly at the two largest catchments. Under high flow conditions, flushing of nitrate was observed, as is typical for stormflow response at Sleepers River. Our field data suggest that in-stream processing of nitrate is likely thermodynamically and kinetically favorable under baseflow but not at higher flow conditions when expanding variable source areas make hydrological connections between nitrate source areas and streams. We are working to evaluate this hypothesis with isotopic and other monitoring data, and to model the coupled interactions of water, DOC, and nitrate fluxes in these nested catchments.

Perchlorate and nitrate treatment by ion exchange integrated with biological brine treatment.

PubMed

Lehman, S Geno; Badruzzaman, Mohammad; Adham, Samer; Roberts, Deborah J; Clifford, Dennis A

2008-02-01

Groundwater contaminated with perchlorate and nitrate was treated in a pilot plant using a commercially available ion exchange (IX) resin. Regenerant brine concentrate from the IX process, containing high perchlorate and nitrate, was treated biologically and the treated brine was reused in IX resin regeneration. The nitrate concentration of the feed water determined the exhaustion lifetime (i.e., regeneration frequency) of the resin; and the regeneration condition was determined by the perchlorate elution profile from the exhausted resin. The biological brine treatment system, using a salt-tolerant perchlorate- and nitrate-reducing culture, was housed in a sequencing batch reactor (SBR). The biological process consistently reduced perchlorate and nitrate concentrations in the spent brine to below the treatment goals of 500 microg ClO4(-)/L and 0.5mg NO3(-)-N/L determined by equilibrium multicomponent IX modeling. During 20 cycles of regeneration, the system consistently treated the drinking water to below the MCL of nitrate (10 mgNO3(-)-N/L) and the California Department of Health Services (CDHS) notification level of perchlorate (i.e., 6 microg/L). A conceptual cost analysis of the IX process estimated that perchlorate and nitrate treatment using the IX process with biological brine treatment to be approximately 20% less expensive than using the conventional IX with brine disposal.

Antarctic polar stratospheric aerosols: The roles of nitrates, chlorides and sulfates

NASA Technical Reports Server (NTRS)

Pueschel, R. F.; Snetsinger, K. G.; Goodman, J. K.; Ferry, G. V.; Oberbeck, V. R.; Verma, S.; Fong, W.

1988-01-01

Nitric and hydrochloric acids have been postulated to condense in the winter polar stratosphere to become an important component of polar stratospheric clouds. One implication is that the removal of NO(y) from the gas phase by this mechanism allows high Cl(x) concentrations to react with O3, because the formation of ClNO3 is inhibited. Contributions of NO3 and Cl to the stratospheric aerosol were determined during the 1987 Airborne Antarctic Ozone Experiment by testing for the presence of nitrates and chlorides in the condensed phase. Aerosol particles were collected on four 500 micron diameter gold wires, each pretreated differently to give results that were specific to certain physical and chemical aerosol properties. One wire was carbon-coated for concentration and size analyses by scanning electron microscopy; X-ray energy dispersive analyses permitted the detection of S and Cl in individual particles. Three more wires were coated with Nitron, barium chloride and silver nitrate, respectively, to detect nitrate, sulfate and chloride in aerosol particles. All three ions, viz., sulfates, nitrates and chlorides were detected in the Antarctic stratospheric aerosol. In terms of number concentrations, the aerosol was dominated by sulfates, followed by chlorides and nitrates. An inverse linear regression can be established between nitrate concentrations and ozone mixing ratio, and between temperature and nitrates.

Soil nitrate reducing processes – drivers, mechanisms for spatial variation, and significance for nitrous oxide production

PubMed Central

Giles, Madeline; Morley, Nicholas; Baggs, Elizabeth M.; Daniell, Tim J.

2012-01-01

The microbial processes of denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are two important nitrate reducing mechanisms in soil, which are responsible for the loss of nitrate (NO3−) and production of the potent greenhouse gas, nitrous oxide (N2O). A number of factors are known to control these processes, including O2 concentrations and moisture content, N, C, pH, and the size and community structure of nitrate reducing organisms responsible for the processes. There is an increasing understanding associated with many of these controls on flux through the nitrogen cycle in soil systems. However, there remains uncertainty about how the nitrate reducing communities are linked to environmental variables and the flux of products from these processes. The high spatial variability of environmental controls and microbial communities across small sub centimeter areas of soil may prove to be critical in determining why an understanding of the links between biotic and abiotic controls has proved elusive. This spatial effect is often overlooked as a driver of nitrate reducing processes. An increased knowledge of the effects of spatial heterogeneity in soil on nitrate reduction processes will be fundamental in understanding the drivers, location, and potential for N2O production from soils. PMID:23264770

Soil nitrate reducing processes - drivers, mechanisms for spatial variation, and significance for nitrous oxide production.

PubMed

Giles, Madeline; Morley, Nicholas; Baggs, Elizabeth M; Daniell, Tim J

2012-01-01

The microbial processes of denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are two important nitrate reducing mechanisms in soil, which are responsible for the loss of nitrate ([Formula: see text]) and production of the potent greenhouse gas, nitrous oxide (N(2)O). A number of factors are known to control these processes, including O(2) concentrations and moisture content, N, C, pH, and the size and community structure of nitrate reducing organisms responsible for the processes. There is an increasing understanding associated with many of these controls on flux through the nitrogen cycle in soil systems. However, there remains uncertainty about how the nitrate reducing communities are linked to environmental variables and the flux of products from these processes. The high spatial variability of environmental controls and microbial communities across small sub centimeter areas of soil may prove to be critical in determining why an understanding of the links between biotic and abiotic controls has proved elusive. This spatial effect is often overlooked as a driver of nitrate reducing processes. An increased knowledge of the effects of spatial heterogeneity in soil on nitrate reduction processes will be fundamental in understanding the drivers, location, and potential for N(2)O production from soils.

Dissimilatory nitrogen reduction in intertidal sediments of a temperate estuary: small scale heterogeneity and novel nitrate-to-ammonium reducers.

PubMed

Decleyre, Helen; Heylen, Kim; Van Colen, Carl; Willems, Anne

2015-01-01

The estuarine nitrogen cycle can be substantially altered due to anthropogenic activities resulting in increased amounts of inorganic nitrogen (mainly nitrate). In the past, denitrification was considered to be the main ecosystem process removing reactive nitrogen from the estuarine ecosystem. However, recent reports on the contribution of dissimilatory nitrate reduction to ammonium (DNRA) to nitrogen removal in these systems indicated a similar or higher importance, although the ratio between both processes remains ambiguous. Compared to denitrification, DNRA has been underexplored for the last decades and the key organisms carrying out the process in marine environments are largely unknown. Hence, as a first step to better understand the interplay between denitrification, DNRA and reduction of nitrate to nitrite in estuarine sediments, nitrogen reduction potentials were determined in sediments of the Paulina polder mudflat (Westerschelde estuary). We observed high variability in dominant nitrogen removing processes over a short distance (1.6 m), with nitrous oxide, ammonium and nitrite production rates differing significantly between all sampling sites. Denitrification occurred at all sites, DNRA was either the dominant process (two out of five sites) or absent, while nitrate reduction to nitrite was observed in most sites but never dominant. In addition, novel nitrate-to-ammonium reducers assigned to Thalassospira, Celeribacter, and Halomonas, for which DNRA was thus far unreported, were isolated, with DNRA phenotype reconfirmed through nrfA gene amplification. This study demonstrates high small scale heterogeneity among dissimilatory nitrate reduction processes in estuarine sediments and provides novel marine DNRA organisms that represent valuable alternatives to the current model organisms.

Dissimilatory nitrogen reduction in intertidal sediments of a temperate estuary: small scale heterogeneity and novel nitrate-to-ammonium reducers

PubMed Central

Decleyre, Helen; Heylen, Kim; Van Colen, Carl; Willems, Anne

2015-01-01

The estuarine nitrogen cycle can be substantially altered due to anthropogenic activities resulting in increased amounts of inorganic nitrogen (mainly nitrate). In the past, denitrification was considered to be the main ecosystem process removing reactive nitrogen from the estuarine ecosystem. However, recent reports on the contribution of dissimilatory nitrate reduction to ammonium (DNRA) to nitrogen removal in these systems indicated a similar or higher importance, although the ratio between both processes remains ambiguous. Compared to denitrification, DNRA has been underexplored for the last decades and the key organisms carrying out the process in marine environments are largely unknown. Hence, as a first step to better understand the interplay between denitrification, DNRA and reduction of nitrate to nitrite in estuarine sediments, nitrogen reduction potentials were determined in sediments of the Paulina polder mudflat (Westerschelde estuary). We observed high variability in dominant nitrogen removing processes over a short distance (1.6 m), with nitrous oxide, ammonium and nitrite production rates differing significantly between all sampling sites. Denitrification occurred at all sites, DNRA was either the dominant process (two out of five sites) or absent, while nitrate reduction to nitrite was observed in most sites but never dominant. In addition, novel nitrate-to-ammonium reducers assigned to Thalassospira, Celeribacter, and Halomonas, for which DNRA was thus far unreported, were isolated, with DNRA phenotype reconfirmed through nrfA gene amplification. This study demonstrates high small scale heterogeneity among dissimilatory nitrate reduction processes in estuarine sediments and provides novel marine DNRA organisms that represent valuable alternatives to the current model organisms. PMID:26528270

Bioreactor performance and functional gene analysis of microbial community in a limited-oxygen fed bioreactor for co-reduction of sulfate and nitrate with high organic input.

PubMed

Xu, Xi-jun; Chen, Chuan; Wang, Ai-jie; Yu, Hao; Zhou, Xu; Guo, Hong-liang; Yuan, Ye; Lee, Duu-jong; Zhou, Jizhong; Ren, Nan-qi

2014-08-15

Limited-oxygen mediated synergistic relationships between sulfate-reducing bacteria (SRB), nitrate-reducing bacteria (NRB) and sulfide-oxidizing bacteria (SOB, including nitrate-reducing, sulfide-oxidizing bacteria NR-SOB) were predicted to simultaneously remove contaminants of nitrate, sulfate and high COD, and eliminate sulfide generation. A lab-scale experiment was conducted to examine the impact of limited oxygen on these oxy-anions degradation, sulfide oxidation and associated microbial functional responses. In all scenarios tested, the reduction of both nitrate and sulfate was almost complete. When limited-oxygen was fed into bioreactors, S(0) formation was significantly improved up to ∼ 70%. GeoChip 4.0, a functional gene microarray, was used to determine the microbial gene diversity and functional potential for nitrate and sulfate reduction, and sulfide oxidation. The diversity of the microbial community in bioreactors was increased with the feeding of limited oxygen. Whereas the intensities of the functional genes involved in sulfate reduction did not show a significant difference, the abundance of the detected denitrification genes decreased in limited oxygen samples. More importantly, sulfide-oxidizing bacteria may alter their populations/genes in response to limited oxygen potentially to function more effectively in sulfide oxidation, especially to elemental sulfur. The genes fccA/fccB from nitrate-reducing, sulfide-oxidizing bacteria (NR-SOB), such as Paracoccus denitrificans, Thiobacillus denitrificans, Beggiatoa sp., Thiomicrospira sp., and Thioalkalivibrio sp., were more abundant under limited-oxygen condition. Copyright © 2014 Elsevier B.V. All rights reserved.

Beetroot and Sodium Nitrate Ameliorate Cardiometabolic Changes in Diet-Induced Obese Hypertensive Rats.

PubMed

Bhaswant, Maharshi; Brown, Lindsay; McAinch, Andrew J; Mathai, Michael L

2017-12-01

Dietary intake of beetroot by humans reduces blood pressure but whether this is caused by nitrate or betanin is not well-defined; neither are effects on other signs of metabolic syndrome. Rats fed a high-carbohydrate, high-fat diet (H) for 16 weeks developed abdominal obesity, hypertension, altered cardiovascular and liver structure and function, and impaired glucose tolerance compared to rats fed a corn starch diet (C). H rats treated with ∼16 mg/kg/day of nitrate either from beetroot juice (H+B) or sodium nitrate (H+N) for the last 8 weeks reduced systolic blood pressure by ∼25 mmHg, improved cardiac structure and function, plasma lipid profile and plasma markers of liver function, reduced inflammatory cell infiltration in heart and liver and decreased left ventricular fibrosis. In the left ventricle, H rats increased mRNA expression of connective tissue growth factor (CTGF), monocyte chemoattractant protein 1 (MCP-1), matrix metalloproteinase-2 (MMP-2), and adenosine monophosphate-activated protein kinase-alpha (AMPK-α) and decreased mRNA expression of peroxisome proliferator-activated receptor-alpha (PPAR-α); both beetroot and sodium nitrate diet-fed rats decreased CTGF threefold, MCP-1, and MMP-2 twofold, and doubled PPAR-α mRNA expression in left ventricular tissue. The similar functional and molecular responses to beetroot and sodium nitrate indicate that the nitrate content of beetroot reduced inflammation and improved cardiovascular, liver, and metabolic function in rats with metabolic syndrome, rather than betanin. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Affinity Purification of Proteins in Tag-Free Form: Split Intein-Mediated Ultrarapid Purification (SIRP).

PubMed

Guan, Dongli; Chen, Zhilei

2017-01-01

Proteins purified using affinity-based chromatography often exploit a recombinant affinity tag. Existing methods for the removal of the extraneous tag, needed for many applications, suffer from poor efficiency and/or high cost. Here we describe a simple, efficient, and potentially low-cost approach-split intein-mediated ultrarapid purification (SIRP)-for both the purification of the desired tagged protein from Escherichia coli lysate and removal of the tag in less than 1 h. The N- and C-fragment of a self-cleaving variant of a naturally split DnaE intein from Nostoc punctiforme are genetically fused to the N-terminus of an affinity tag and a protein of interest (POI), respectively. The N-intein/affinity tag is used to functionalize an affinity resin. The high affinity between the N- and C-fragment of DnaE intein enables the POI to be purified from the lysate via affinity to the resin, and the intein-mediated C-terminal cleavage reaction causes tagless POI to be released into the flow-through. The intein cleavage reaction is strongly inhibited by divalent ions (e.g., Zn 2+ ) under non-reducing conditions and is significantly enhanced by reducing conditions. The POI is cleaved efficiently regardless of the identity of the N-terminal amino acid except in the cases of threonine and proline, and the N-intein-functionalized affinity resin can be regenerated for multiple cycles of use.

Determination of intracellular nitrate.

PubMed Central

Romero, J M; Lara, C; Guerrero, M G

1989-01-01

A sensitive procedure has been developed for the determination of intracellular nitrate. The method includes: (i) preparation of cell lysates in 2 M-H3PO4 after separation of cells from the outer medium by rapid centrifugation through a layer of silicone oil, and (ii) subsequent nitrate analysis by ion-exchange h.p.l.c. with, as mobile phase, a solution containing 50 mM-H3PO4 and 2% (v/v) tetrahydrofuran, adjusted to pH 1.9 with NaOH. The determination of nitrate is subjected to interference by chloride and sulphate when present in the samples at high concentrations. Nitrite also interferes, but it is easily eliminated by treatment of the samples with sulphamic acid. The method has been successfully applied to the study of nitrate transport in the unicellular cyanobacterium Anacystis nidulans. PMID:2497740

Measurement of trace nitrate concentrations in seawater by ion chromatography with valve switching

NASA Astrophysics Data System (ADS)

Du, Juan; Fa, Yun; Zheng, Yue; Li, Xuebing; Du, Fanglin; Yang, Haiyan

2014-05-01

An ion chromatographic method with a valve switching facility was developed to determine trace nitrate concentrations in seawater using two pumps, two different suppressors, and two columns. A carbohydrate membrane desalter was used to reduce the high concentrations of sodium salts in samples. In this method, trace nitrate was eluted from the concentrator column to the analytical columns, while the matrix fl owed to waste. Neither chemical pre-treatment nor sample dilution was required. In the optimized separation conditions, the method showed good linearity ( R >0.99) in the 0.05 and 50 mg/L concentration range, and satisfactory repeatability (RSD<5%, n =6). The limit of detection for nitrate was 0.02 mg/L. Results showed that the valve switching system was suitable and practical for the determination of trace nitrate in seawater.

Spherical nitroguanidine process

DOEpatents

Sanchez, John A.; Roemer, Edward L.; Stretz, Lawrence A.

1990-01-01

A process of preparing spherical high bulk density nitroguanidine by dissing low bulk density nitroguanidine in N-methyl pyrrolidone at elevated temperatures and then cooling the solution to lower temperatures as a liquid characterized as a nonsolvent for the nitroguanidine is provided. The process is enhanced by inclusion in the solution of from about 1 ppm up to about 250 ppm of a metal salt such as nickel nitrate, zinc nitrate or chromium nitrate, preferably from about 20 to about 50 ppm.

Nitrate reduction in a simulated free-water surface wetland system.

PubMed

Misiti, Teresa M; Hajaya, Malek G; Pavlostathis, Spyros G

2011-11-01

The feasibility of using a constructed wetland for treatment of nitrate-contaminated groundwater resulting from the land application of biosolids was investigated for a site in the southeastern United States. Biosolids degradation led to the release of ammonia, which upon oxidation resulted in nitrate concentrations in the upper aquifer in the range of 65-400 mg N/L. A laboratory-scale system was constructed in support of a pilot-scale project to investigate the effect of temperature, hydraulic retention time (HRT) and nitrate and carbon loading on denitrification using soil and groundwater from the biosolids application site. The maximum specific reduction rates (MSRR), measured in batch assays conducted with an open to the atmosphere reactor at four initial nitrate concentrations from 70 to 400 mg N/L, showed that the nitrate reduction rate was not affected by the initial nitrate concentration. The MSRR values at 22 °C for nitrate and nitrite were 1.2 ± 0.2 and 0.7 ± 0.1 mg N/mg VSS(COD)-day, respectively. MSRR values were also measured at 5, 10, 15 and 22 °C and the temperature coefficient for nitrate reduction was estimated at 1.13. Based on the performance of laboratory-scale continuous-flow reactors and model simulations, wetland performance can be maintained at high nitrogen removal efficiency (>90%) with an HRT of 3 days or higher and at temperature values as low as 5 °C, as long as there is sufficient biodegradable carbon available to achieve complete denitrification. The results of this study show that based on the climate in the southeastern United States, a constructed wetland can be used for the treatment of nitrate-contaminated groundwater to low, acceptable nitrate levels. Copyright © 2011 Elsevier Ltd. All rights reserved.

Drivers of inverse DOC-nitrate loss patterns in forest soils and streams

NASA Astrophysics Data System (ADS)

Goodale, C. L.

2013-12-01

Nitrate loss from forested catchments varies greatly across sites and over time, with few reliable correlates. One of the few recurring patterns, however, is the negative nonlinear relationship that occurs regularly between surface water nitrate and dissolved organic carbon (DOC) concentrations: that is, nitrate declines sharply as DOC concentrations increase, and high nitrate levels occur only at low DOC concentrations. Several hypotheses have been proposed to explain this pattern, but its cause has remained speculative. It is likely to be driven by C- or N-limitation of biological processes such as assimilation or denitrification, but the identity of which biological process or the main landscape position of their activity are not known. We examined whether DOC and nitrate are both driven by soil C content, at scales of both soil blocks and across catchments, by measuring soil, soil extract, and surface water chemistry across nine catchments selected from long-term monitoring networks in the Catskill and Adirondack Mountains. We measured soil C and N status and solution nitrate, DOC, bioavailable DOC (bDOC), and isotopic composition (13C-DOC, 15N- and 18O-NO3) to examine whether variation in stocks of soil C partly controls DOC and nitrate loss from forested catchments in New York State. These measurements showed that surface soil C and C:N ratio together determine soil production of DOC and nitrate, reflecting assimilative demand for N by heterotrophic microbes. Yet, they also show that these processes do not produce the inverse DOC-NO3 curve observed at the catchment scale. Rather, catchment-scale DOC-nitrate patterns are more likely to be governed by the balance between excess nitrate production and its bDOC-mediated loss to denitrification.

Nitrate-induced genes in tomato roots. Array analysis reveals novel genes that may play a role in nitrogen nutrition.

PubMed

Wang, Y H; Garvin, D F; Kochian, L V

2001-09-01

A subtractive tomato (Lycopersicon esculentum) root cDNA library enriched in genes up-regulated by changes in plant mineral status was screened with labeled mRNA from roots of both nitrate-induced and mineral nutrient-deficient (-nitrogen [N], -phosphorus, -potassium [K], -sulfur, -magnesium, -calcium, -iron, -zinc, and -copper) tomato plants. A subset of cDNAs was selected from this library based on mineral nutrient-related changes in expression. Additional cDNAs were selected from a second mineral-deficient tomato root library based on sequence homology to known genes. These selection processes yielded a set of 1,280 mineral nutrition-related cDNAs that were arrayed on nylon membranes for further analysis. These high-density arrays were hybridized with mRNA from tomato plants exposed to nitrate at different time points after N was withheld for 48 h, for plants that were grown on nitrate/ammonium for 5 weeks prior to the withholding of N. One hundred-fifteen genes were found to be up-regulated by nitrate resupply. Among these genes were several previously identified as nitrate responsive, including nitrate transporters, nitrate and nitrite reductase, and metabolic enzymes such as transaldolase, transketolase, malate dehydrogenase, asparagine synthetase, and histidine decarboxylase. We also identified 14 novel nitrate-inducible genes, including: (a) water channels, (b) root phosphate and K(+) transporters, (c) genes potentially involved in transcriptional regulation, (d) stress response genes, and (e) ribosomal protein genes. In addition, both families of nitrate transporters were also found to be inducible by phosphate, K, and iron deficiencies. The identification of these novel nitrate-inducible genes is providing avenues of research that will yield new insights into the molecular basis of plant N nutrition, as well as possible networking between the regulation of N, phosphorus, and K nutrition.

Evaluating the source of streamwater nitrate using δ15N and δ18O in nitrate in two watersheds in New Hampshire, USA

USGS Publications Warehouse

Pardo, Linda H.; Kendall, Carol; Pett-Ridge, Jennifer; Chang, Cecily C.Y.

2004-01-01

The natural abundance of nitrogen and oxygen isotopes in nitrate can be a powerful tool for identifying the source of nitrate in streamwater in forested watersheds, because the two main sources of nitrate, atmospheric deposition and microbial nitrification, have distinct δ18O values. Using a simple mixing model, we estimated the relative fractions in streamwater derived from these sources for two forested watersheds with markedly different streamwater nitrate outputs. In this study, we monitored δ15N and δ18O of nitrate biweekly in atmospheric deposition and in streamwater for 20 months at the Hubbard Brook Experimental Forest, New Hampshire, USA (moderate nitrogen export), and monthly in streamwater at the Bowl Research Natural Area, New Hampshire, USA (high nitrogen export). For rain, δ18O values ranged from +47 to +77‰ (mean: +58‰) and δ15N from −5 to +1‰ (mean: −3‰); for snow, δ18O values ranged from +52 to +75‰ (mean: +67‰) and δ15N from −3 to +2‰ (mean: −1‰). Streamwater nitrate, in contrast to deposition, had δ18O values between +12 and +33‰ (mean: +18‰) and δ15N between −3 and +6‰ (mean: 0‰). Since nitrate produced by nitrification typically has δ18O values ranging from −5 to +15‰, our field data suggest that most of the nitrate lost from the watersheds in streamflow was nitrified within the catchment. Our results confirm the importance of microbial nitrogen transformations in regulating nitrogen losses from forested ecosystems and suggest that hydrologic storage may be a factor in controlling catchment nitrate losses.

Impacts of management and climate change on nitrate leaching in a forested karst area.

PubMed

Dirnböck, Thomas; Kobler, Johannes; Kraus, David; Grote, Rüdiger; Kiese, Ralf

2016-01-01

Forest management and climate change, directly or indirectly, affect drinking water resources, both in terms of quality and quantity. In this study in the Northern Limestone Alps in Austria we have chosen model calculations (LandscapeDNDC) in order to resolve the complex long-term interactions of management and climate change and their effect on nitrogen dynamics, and the consequences for nitrate leaching from forest soils into the karst groundwater. Our study highlights the dominant role of forest management in controlling nitrate leaching. Both clear-cut and shelterwood-cut disrupt the nitrogen cycle to an extent that causes peak concentrations and high fluxes into the seepage water. While this effect is well known, our modelling approach has revealed additional positive as well as negative impacts of the expected climatic changes on nitrate leaching. First, we show that peak nitrate concentrations during post-cutting periods were elevated under all climate scenarios. The maximal effects of climatic changes on nitrate concentration peaks were 20-24 mg L(-1) in 2090 with shelterwood or clear-cut management. Second, climate change significantly decreased the cumulative nitrate losses over full forest rotation periods (by 10-20%). The stronger the expected temperature increase and precipitation decrease (in summer), the lesser were the observed nitrate losses. However, mean annual seepage water nitrate concentrations and cumulative nitrate leaching were higher under continuous forest cover management than with shelterwood-cut and clear-cut systems. Watershed management can thus be adapted to climate change by either reducing peak concentrations or long-term loads of nitrate in the karst groundwater. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of temperature & salt concentration on salt tolerant nitrate-perchlorate reducing bacteria: Nitrate degradation kinetics.

PubMed

Ebrahimi, Shelir; Nguyen, Thi Hau; Roberts, Deborah J

2015-10-15

The sustainability of nitrate-contaminated water treatment using ion-exchange processes can be achieved by regenerating the exhausted resin several times. Our previous study shows that the use of multi-cycle bioregeneration of resin enclosed in membrane is an effective and innovative regeneration method. In this research, the effects of two independent factors (temperature and salt concentration) on the biological denitrification rate were studied. The results of this research along with the experimental results of the previous study on the effect of the same factors on nitrate desorption rate from the resin allow the optimization of the bioregeneration process. The results of nitrate denitrification rate study show that the biodegradation rate at different temperature and salt concentration is independent of the initial nitrate concentration. At each specific salt concentration, the nitrate removal rate increased with increasing temperature with the average value of 0.001110 ± 0.0000647 mg-nitrate/mg-VSS.h.°C. However, the effect of different salt concentrations was dependent on the temperature; there is a significant interaction between salt concentration and temperature; within each group of temperatures, the nitrate degradation rate decreased with increasing the salt concentration. The temperature affected the tolerance to salinity and culture was less tolerant to high concentration of salt at low temperature. Evidenced by the difference between the minimum and maximum nitrate degradation rate being greater at lower temperature. At 35 °C, a 32% reduction in the nitrate degradation rate was observed while at 12 °C this reduction was 69%. This is the first published study to examine the interaction of salt concentration and temperature during biological denitrification. Copyright © 2015 Elsevier Ltd. All rights reserved.

Village-Level Identification of Nitrate Sources: Collaboration of Experts and Local Population in Benin, Africa

NASA Astrophysics Data System (ADS)

Crane, P.; Silliman, S. E.; Boukari, M.; Atoro, I.; Azonsi, F.

2005-12-01

Deteriorating groundwater quality, as represented by high nitrates, in the Colline province of Benin, West Africa was identified by the Benin national water agency, Direction Hydraulique. For unknown reasons the Colline province had consistently higher nitrate levels than any other region of the country. In an effort to address this water quality issue, a collaborative team was created that incorporated professionals from the Universite d'Abomey-Calavi (Benin), the University of Notre Dame (USA), Direction l'Hydraulique (a government water agency in Benin), Centre Afrika Obota (an educational NGO in Benin), and the local population of the village of Adourekoman. The goals of the project were to: (i) identify the source of nitrates, (ii) test field techniques for long term, local monitoring, and (iii) identify possible solutions to the high levels of groundwater nitrates. In order to accomplish these goals, the following methods were utilized: regional sampling of groundwater quality, field methods that allowed the local population to regularly monitor village groundwater quality, isotopic analysis, and sociological methods of surveys, focus groups, and observations. It is through the combination of these multi-disciplinary methods that all three goals were successfully addressed leading to preliminary identification of the sources of nitrates in the village of Adourekoman, confirmation of utility of field techniques, and initial assessment of possible solutions to the contamination problem.

Effects of Parental Temperature and Nitrate on Seed Performance are Reflected by Partly Overlapping Genetic and Metabolic Pathways.

PubMed

He, Hanzi; Willems, Leo A J; Batushansky, Albert; Fait, Aaron; Hanson, Johannes; Nijveen, Harm; Hilhorst, Henk W M; Bentsink, Leónie

2016-03-01

Seed performance is affected by the seed maturation environment, and previously we have shown that temperature, nitrate and light intensity were the most influential environmental factors affecting seed performance. Seeds developed in these environments were selected to assess the underlying metabolic pathways, using a combination of transcriptomics and metabolomics. These analyses revealed that the effects of the parental temperature and nitrate environments were reflected by partly overlapping genetic and metabolic networks, as indicated by similar changes in the expression levels of metabolites and transcripts. Nitrogen metabolism-related metabolites (asparagine, γ-aminobutyric acid and allantoin) were significantly decreased in both low temperature (15 °C) and low nitrate (N0) maturation environments. Correspondingly, nitrogen metabolism genes (ALLANTOINASE, NITRATE REDUCTASE 1, NITRITE REDUCTASE 1 and NITRILASE 4) were differentially regulated in the low temperature and nitrate maturation environments, as compared with control conditions. High light intensity during seed maturation increased galactinol content, and displayed a high correlation with seed longevity. Low light had a genotype-specific effect on cell surface-encoding genes in the DELAY OF GERMINATION 6-near isogenic line (NILDOG6). Overall, the integration of phenotypes, metabolites and transcripts led to new insights into the regulation of seed performance. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

The fourth dimension in immunological space: how the struggle for nutrients selects high-affinity lymphocytes.

PubMed

Wensveen, Felix M; van Gisbergen, Klaas P J M; Eldering, Eric

2012-09-01

Lymphocyte activation via the antigen receptor is associated with radical shifts in metabolism and changes in requirements for nutrients and cytokines. Concomitantly, drastic changes occur in the expression of pro-and anti-apoptotic proteins that alter the sensitivity of lymphocytes to limiting concentrations of key survival factors. Antigen affinity is a primary determinant for the capacity of activated lymphocytes to access these vital resources. The shift in metabolic needs and the variable access to key survival factors is used by the immune system to eliminate activated low-affinity cells and to generate an optimal high-affinity response. In this review, we focus on the control of apoptosis regulators in activated lymphocytes by nutrients, cytokines, and costimulation. We propose that the struggle among individual clones that leads to the formation of high-affinity effector cell populations is in effect an 'invisible' fourth signal required for effective immune responses. © 2012 John Wiley & Sons A/S.

Role of framework mutations and antibody flexibility in the evolution of broadly neutralizing antibodies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ovchinnikov, Victor; Louveau, Joy E.; Barton, John P.

Eliciting antibodies that are cross reactive with surface proteins of diverse strains of highly mutable pathogens (e.g., HIV, influenza) could be key for developing effective universal vaccines. Mutations in the framework regions of such broadly neutralizing antibodies (bnAbs) have been reported to play a role in determining their properties. We used molecular dynamics simulations and models of affinity maturation to study specific bnAbs against HIV. Our results suggest that there are different classes of evolutionary lineages for the bnAbs. If germline B cells that initiate affinity maturation have high affinity for the conserved residues of the targeted epitope, framework mutationsmore » increase antibody rigidity as affinity maturation progresses to evolve bnAbs. If the germline B cells exhibit weak/moderate affinity for conserved residues, an initial increase in flexibility via framework mutations may be required for the evolution of bnAbs. Subsequent mutations that increase rigidity result in highly potent bnAbs. Implications of our results for immunogen design are discussed.« less

Role of framework mutations and antibody flexibility in the evolution of broadly neutralizing antibodies

DOE PAGES

Ovchinnikov, Victor; Louveau, Joy E.; Barton, John P.; ...

2018-02-14

Eliciting antibodies that are cross reactive with surface proteins of diverse strains of highly mutable pathogens (e.g., HIV, influenza) could be key for developing effective universal vaccines. Mutations in the framework regions of such broadly neutralizing antibodies (bnAbs) have been reported to play a role in determining their properties. We used molecular dynamics simulations and models of affinity maturation to study specific bnAbs against HIV. Our results suggest that there are different classes of evolutionary lineages for the bnAbs. If germline B cells that initiate affinity maturation have high affinity for the conserved residues of the targeted epitope, framework mutationsmore » increase antibody rigidity as affinity maturation progresses to evolve bnAbs. If the germline B cells exhibit weak/moderate affinity for conserved residues, an initial increase in flexibility via framework mutations may be required for the evolution of bnAbs. Subsequent mutations that increase rigidity result in highly potent bnAbs. Implications of our results for immunogen design are discussed.« less

Role of framework mutations and antibody flexibility in the evolution of broadly neutralizing antibodies

PubMed Central

2018-01-01

Eliciting antibodies that are cross reactive with surface proteins of diverse strains of highly mutable pathogens (e.g., HIV, influenza) could be key for developing effective universal vaccines. Mutations in the framework regions of such broadly neutralizing antibodies (bnAbs) have been reported to play a role in determining their properties. We used molecular dynamics simulations and models of affinity maturation to study specific bnAbs against HIV. Our results suggest that there are different classes of evolutionary lineages for the bnAbs. If germline B cells that initiate affinity maturation have high affinity for the conserved residues of the targeted epitope, framework mutations increase antibody rigidity as affinity maturation progresses to evolve bnAbs. If the germline B cells exhibit weak/moderate affinity for conserved residues, an initial increase in flexibility via framework mutations may be required for the evolution of bnAbs. Subsequent mutations that increase rigidity result in highly potent bnAbs. Implications of our results for immunogen design are discussed. PMID:29442996

Affinity Crystallography: A New Approach to Extracting High-Affinity Enzyme Inhibitors from Natural Extracts.

PubMed

Aguda, Adeleke H; Lavallee, Vincent; Cheng, Ping; Bott, Tina M; Meimetis, Labros G; Law, Simon; Nguyen, Nham T; Williams, David E; Kaleta, Jadwiga; Villanueva, Ivan; Davies, Julian; Andersen, Raymond J; Brayer, Gary D; Brömme, Dieter

2016-08-26

Natural products are an important source of novel drug scaffolds. The highly variable and unpredictable timelines associated with isolating novel compounds and elucidating their structures have led to the demise of exploring natural product extract libraries in drug discovery programs. Here we introduce affinity crystallography as a new methodology that significantly shortens the time of the hit to active structure cycle in bioactive natural product discovery research. This affinity crystallography approach is illustrated by using semipure fractions of an actinomycetes culture extract to isolate and identify a cathepsin K inhibitor and to compare the outcome with the traditional assay-guided purification/structural analysis approach. The traditional approach resulted in the identification of the known inhibitor antipain (1) and its new but lower potency dehydration product 2, while the affinity crystallography approach led to the identification of a new high-affinity inhibitor named lichostatinal (3). The structure and potency of lichostatinal (3) was verified by total synthesis and kinetic characterization. To the best of our knowledge, this is the first example of isolating and characterizing a potent enzyme inhibitor from a partially purified crude natural product extract using a protein crystallographic approach.

Predicting the dynamics of bacterial growth inhibition by ribosome-targeting antibiotics

NASA Astrophysics Data System (ADS)

Greulich, Philip; Doležal, Jakub; Scott, Matthew; Evans, Martin R.; Allen, Rosalind J.

2017-12-01

Understanding how antibiotics inhibit bacteria can help to reduce antibiotic use and hence avoid antimicrobial resistance—yet few theoretical models exist for bacterial growth inhibition by a clinically relevant antibiotic treatment regimen. In particular, in the clinic, antibiotic treatment is time-dependent. Here, we use a theoretical model, previously applied to steady-state bacterial growth, to predict the dynamical response of a bacterial cell to a time-dependent dose of ribosome-targeting antibiotic. Our results depend strongly on whether the antibiotic shows reversible transport and/or low-affinity ribosome binding (‘low-affinity antibiotic’) or, in contrast, irreversible transport and/or high affinity ribosome binding (‘high-affinity antibiotic’). For low-affinity antibiotics, our model predicts that growth inhibition depends on the duration of the antibiotic pulse, and can show a transient period of very fast growth following removal of the antibiotic. For high-affinity antibiotics, growth inhibition depends on peak dosage rather than dose duration, and the model predicts a pronounced post-antibiotic effect, due to hysteresis, in which growth can be suppressed for long times after the antibiotic dose has ended. These predictions are experimentally testable and may be of clinical significance.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beaumont, K.; Vaughn, D.A.; Fanestil, D.D.

Thiazides and related diuretics inhibit NaCl reabsorption in the distal tubule through an unknown mechanism. The authors report here that ({sup 3}H)metolazone, a diuretic with a thiazide-like mechanism of action, labels a site in rat kidney membranes that has characteristics of the thiazide-sensitive ion transporter. ({sup 3}H)Metolazone bound with high affinity to a site with a density of 0.717 pmol/mg of protein in kidney membranes. The binding site was localized to the renal cortex, with little or not binding in other kidney regions and 11 other tissues. The affinities of thiazide-type diuretics for this binding site were significantly correlated withmore » their clinical potency. Halide anions specifically inhibited high-affinity binding of ({sup 3}H)metolazone to this site. ({sup 3})Metolazone also bound with lower affinity to sites present in kidney as well as in liver, testis, lung, brain, heart, and other tissues. Calcium antagonists and certain smooth muscle relaxants had K{sub i} values of 0.6-10 {mu}M for these low-affinity sites, which were not inhibited by most of the thiazide diuretics tested. Properties of the high-affinity ({sup 3}H)metolazone binding site are consistent with its identity as the receptor for thiazide-type diuretics.« less