Mutagenicity of the reaction products of dibenzo-p-dioxin with nitrogen oxides.

Dibenzo-p-dioxin (DD) was made to react with various concentrations of nitrogen oxides in the dark. The mutagenicities of the reaction products were tested using Salmonella typhimurium strains TA98, TA100, TA98NR and TA98/1,8-DNP6 in the presence or absence of a mammalian metabolic activation system (S9 mix). DD-NOx (molar ratios 1:3, 1:6 and 1:18) reaction products exhibited mutagenic potency in strains TA98 and TA98/1,8-DNP6 without S9 mix. In a gas chromatography/mass spectrometry study, 2-nitrodibenzo-p-dioxin (NDD) was identified with authentic sample in the mutagenic reaction products. DD-NOx (1:18) reaction products were reduced by sodium hydrogen sulfide and the reduction mixture was analyzed by HPLC. 2,7-Dinitrodibenzo-p-dioxin (DNDD) and 2,8-DNDD were identified as corresponding diamino-DDs in the reduction mixture. 2-NDD, 2,7-DNDD and 2,8-DNDD were also mutagenic in strains TA98 and TA98/1,8-DNP6 without S9 mix and the mutagenicity of DD-NOx reaction products was largely accounted for by the nitro-DDs.     

Regulation of prostaglandin H2 synthase activity by nitrogen oxides.

Nitric oxide and its derivatives have been shown to both activate and inhibit prostaglandin H(2) synthase 1 (PGHS-1). We set out to determine the mechanisms by which different nitrogen oxide derivatives modulate PGHS-1 activity. To this end, we show that 3-morpholinosydnonimine hydrochloride (SIN-1), a compound capable of generating peroxynitrite, activates purified PGHS-1 and also stimulates PGE(2) production in arterial smooth muscle cells in the presence of exogenous arachidonic acid. The effect of SIN-1 in smooth muscle cells was abrogated by superoxide and peroxynitrite inhibitors, which supports the hypothesis that peroxynitrite is an activating species of PGHS-1. Indeed, authentic peroxynitrite also induced PGE(2) production in arachidonic acid-stimulated cells. In contrast, when cells were exposed to the nitric oxide-releasing compound 1-hydroxy-2-oxo-3-[(methylamino)propyl]-3-methyl-1-triazene (NOC-7), PGHS-1 enzyme activity was inhibited in the presence of exogenous arachidonic acid. Finally, in lipid-loaded smooth muscle cells, we demonstrate that SIN-1 stimulates arachidonic acid-induced PGE(2) production; albeit, the extent of activation is reduced compared to that under normal conditions. These results indicate that formation of peroxynitrite is a key intermediary step in PGHS-1 activation. However, other forms of NO(x)() inhibit PGHS-1. These results may have implications in the regulation of vascular function and tone in normal and atherosclerotic arteries.     

Transformations of 2,6-diisopropylphenol by NO-derived nitrogen oxides, particularly peroxynitrite.

To investigate the protective effect of the anesthetic 2, 6-diisopropylphenol, or propofol, in oxidative processes in which (*)NO and peroxynitrite are involved, direct interactions were explored. The reactions of the highly lipophilic propofol with (*)NO in methanolic or aqueous buffered solutions under air were shown to produce the same compounds as those detected with peroxynitrite, but with very low yields and slow rates. In aqueous neutral medium, peroxynitrite (ONOO(-), ONOOCO(-)(2), ONOOH) was able to nitrate and oxidize propofol: In addition to oxidation products, quinone and quinone dimer, the formation of the 4-nitropropofol derivative was detected, increasing with peroxynitrite or CO(2) concentrations. Nitration reached 20% after the addition of 25 mM bicarbonate to an equimolecular mixture of peroxynitrite and propofol in methanol/phosphate-buffered solution (1/4,v/v) at pH 7.4. However, peroxynitrite either in methanol or in alkaline-buffered mixture (optimum pH 10-12) resulted in the rapid and almost complete transformation of propofol to an intermediate compound 1, which further decomposed to 4-nitrosopropofol. The transient compound 1 was obtained from either peroxynitrite or (*)NO in the presence of oxygen. From mass spectrometry determination of compound 1 we propose the involvement of the nitrosodioxyl radical ONOO(*), forming an adduct with the propofoxyl radical, to yield 4-nitrosodioxypropofol and finally 4-nitrosopropofol.     

Occurrence of nitric and nitrous oxides in a coastal marine sediment.

The distribution of denitrification activity in a coastal marine sediment was determined by the acetylene inhibition technique and compared to concentration profiles of NO3-, NO2-, NO, and N2O. The bulk of the denitrification activity was associated with the accumulation of NO3- in the oxidized surface zone of the sediment, but a secondary denitrification zone was occasionally found in the deeper layers where oxidized patches had been introduced by the burrowing activity of the macrofauna. Maxima of NO and N2O were not associated with the peak activity of denitrification in the surface zone but were located at the lower edge of the activity profile. Significant accumulation of NO was found at the redox transition zone towards the deeper, sulfide-rich layers.     

Preparation and enzymatic recognition of guanine lesions induced by nitrogen oxides.

Xanthine (Xan) and oxanine (Oxa) are the major deamination products of guanine formed by the treatment with nitrogen oxides (e.g., NO and HNO2). In this study, 2'-deoxyribonucleoside 5'-triphosphates of Xan and Oxa were prepared by the NaNO2 treatment of dGTP. These modified nucleotides were incorporated into oligonucleotides by DNA polymerase reactions. The repair activities of various DNA N-glycosylases for Xan and Oxa were examined using these substrates.     

Gram-scale chromatographic purification of beta-sitosterol. Synthesis and characterization of beta-sitosterol oxides

An effective purification method for beta-sitosterol was developed starting from a commercial source of a phytosterol mixture using preparative adsorption column chromatography. beta-Sitosterol (> or = 95% purity) was obtained on a gram-scale. Thus, the synthesis of six beta-sitosterol oxides, including 7alpha-hydroxy, 7beta-hydroxy, 5,6alpha-epoxy, 5,6beta-epoxy, 7-keto, and 5alpha,6beta-dihydroxysitosterol, were successfully carried out. The spectral characteristics of all the synthetic intermediates and target compounds (approximately 95% purity) were well-documented.     

Fungal transformations of uranium oxides

The biogeochemical activities of free-living and symbiotic fungi must be acknowledged in attempts to understand uranium cycling and dispersal in the environment. Although the near-surface geochemistry of uranium is very complex and a wide variety of mineral phases is known, uranium trioxide (UO3) and triuranium octaoxide (U(3)O(8)) can be used as well characterized models in the study of biotransformations. We have used a complex methodological approach involving advanced solid state speciation and scanning electron microscopy to study the ability of saprotrophic, ericoid and ectomycorrhizal fungi to transform these model oxides. This study has revealed that fungi exhibit a high uranium oxide tolerance, and possess the ability to solubilize UO3 and U(3)O(8) and to accumulate uranium within the mycelium to over 80 mg (g dry weight)(-1) biomass. X-ray absorption spectroscopy of uranium speciation within the biomass showed that in most of the fungi the uranyl ion was coordinated to phosphate ligands, but in ectomycorrhizal fungi mixed phosphate/carboxylate coordination was observed. Abundant uranium precipitates associated with phosphorus were found in the mycelium and encrusted the hyphae. Some of the fungi caused the biomineralization of well-crystallized uranyl phosphate minerals of the meta-autunite group. This is the first experimental evidence for fungal transformations of uranium solids and the production of secondary mycogenic uranium minerals.     

The preparation of tin-molybdenum oxides

The characterisation of ‘tin-molybdenum oxides’ prepared by a variety of methods has been performed by X-ray diffraction. The results show that preparative procedures involving the calcination of precipitates or evaporates containing the two cations at high temperatures are more likely to result in solid solution formation than methods which involve the calcination of slurries or solid state reactions. However, the monophasic tin-molybdenum oxides formed by the moderate temperature calcination of precipitates or evaporates undergo further changes at elevated temperatures which indicate that bulk equilibrium is difficult to achieve in this ternary system.     

Reductive dissolution of Fe(III) oxides by Pseudomonas sp. 200

The kinetics and mechanism of reductive dissolution of Fe(III) oxides were examined in pure, batch cultures of Pseudomonassp. 200. Primary factors controlling hematite dissolution kinetics were mineral surface area (or concentration of high-energy surface sites), ligand concentration, and cell number. In the presence of nitrilotriacetic acid (NTA), saturation kinetics were apparent in the relationship governing reductive dissolution of hematite. A kinetic expression was developed in which overall iron-reduction rate is functionally related to the concentrations of both NTA and Fe(III).Addition of NTA resulted in a 20-fold increase in the microbial rate of mineral (reductive) dissolution. Mechanisms in which NTA served as a bridging ligand, shuttling respiratory electrons from the membrane-bound microbial electron transport chain to the metal center of the iron oxide, or accelerated the departure of Fe(II) centers (bound to ligand) from the oxide surface following reduction have been postulated. Experimental results indicated that cell-mineral contact was essential for reductive dissolution of goethite.     

Solubility and dissolution of iron oxides

In most soils, FeIII oxides (group name) are the common source of Fe for plant nutrition. Since this Fe has to be supplied via solution, the solubility and the dissolution rate of the Fe oxides are essential for the Fe supply. Hydrolysis constants and solubility products (K_sp) describing the effect of pH on FeIII ion concentration in solution are available for the well-known Fe oxides occurring in soils such as goethite, hematite, ferrihydrite. K_sp values are usually extremely low ((Fe³⁺)·(OH)³=10^–37–10^–44). However, for each mineral type, K_sp may increase by several orders of magnitude with decreasing crystal size and it decreases with increasing Al substitution assuming ideal solid solution between the pure end-members. Based on such calculations a poorly crystalline goethite with a crystal size of 5 nm may well reach the solubility of ferrihydrite. The variations in K_sp are of relevance for soils because crystal size and Al substitution of soil Fe oxides vary considerably and can now be determined relatively easily.The concentration of Fe²⁺ in soil solutions is often much higher than that of Fe(III) ions. Therefore, redox potential strongly influences the activity of FeII. At a given pH and E_h, the activity of FeII is higher the higher K_sp of the FeIII oxide and thus also varies with the type of Fe oxide present.Besides the solubility, it is the dissolution rate which governs the supply of soluble Fe to the plant roots. Dissolution of Fe oxides takes place either by protonation, complexation or, most important, by reduction. Numerous dissolution rate studies with various FeIII oxides were conducted in strong mineral acids (protonation) and they have shown that besides the Fe oxide species, crystal size and/or crystal order and substitution are important determinative factors. For example, in soils, small amounts of a more highly soluble meta- or instable Fe oxide such as ferrihydrite with a large specific surface (several hundred m²g^–1) may be essential for the Fe supply to the plant root. Its higher dissolution rate can also be used to quantify its amount in soils. Ferrihydrite can be an important component in soils with high amounts of organic matter and/or active redox dynamics, whereas highly aerated and strongly weathered soils are usually very low in ferrihydrite. On the other hand, dissolution rates of goethites decrease as their Al substitution increases.Much less information exists on the rate of reductive and chelative dissolution of Fe oxides which generally simulate soil conditions better than dissolution by protonation. Here again, type of oxide, crystal size and substitution are important factors. Organic anions such as oxalate, which are adsorbed at the surface, may weaken the Fe³⁺-O bonds and thereby increase reductive dissolution. As often observed in weathering, the dissolution features of the crystals appear to follow zones of weakness in the crystal.     

Microperoxidase 8 catalysed nitrogen oxides formation from oxidation of N-hydroxyguanidines by hydrogen peroxide.

Nitric oxide (NO) is a potent intra- and intercellular messenger involved in the control of vascular tone, neuronal signalling and host response to infection. In mammals, NO is synthesized by oxidation of l-arginine catalysed by hemeproteins called NO-synthases with intermediate formation of Nomega-hydroxy-l-arginine (NOHA). NOHA and some hydroxyguanidines have been shown to be able to deliver nitrogen oxides including NO in the presence of various oxidative systems. In this study, NOHA and a model compound, N-(4-chlorophenyl)-N'-hydroxyguanidine, were tested for their ability to generate NO in the presence of a haemprotein model, microperoxidase 8 (MP8), and hydrogen peroxide. Nitrite and nitrate production along with selective formation of 4-chlorophenylcyanamide was observed from incubations of N-(4-chlorophenyl)-N'-hydroxyguanidine in the presence of MP8 and hydrogen peroxide. In the case of NOHA, the corresponding cyanamide, Ndelta-cyano-L-ornithine, was too unstable under the conditions used and l-citrulline was the only product identified. A NO-specific conversion of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide to 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl and formation of MP8-Fe-NO complexes were observed by EPR spectroscopy and low-temperature UV/visible spectroscopy, respectively. These results clearly demonstrate the formation of nitrogen oxides including NO from the oxidation of exogenous hydroxyguanidines by hydrogen peroxide in the presence of a minienzyme such as MP8. The importance of the bioactivation of endogenous (NOHA) or exogenous N-hydroxyguanidines by peroxidases of physiological interest remains to be established in vivo.     

Efficacy of zinc oxides as fertilisers

Background and aims

Human zinc (Zn) deficiency is prevalent in developing countries and Zn biofortification of grains is used to increase the Zn content of food staples. Agronomic interventions to biofortify grain involve fertiliser selection and management. The usefulness of a zinc compound as a fertiliser will depend on its solubility, bioavailability and the effect of its distribution in the soil profile.

Methods

Various sources of Zn oxide and Zn sulfate fertiliser were characterised for nutrient content, morphology, solubility, and fertiliser recovery when applied to the surface, banded near the seed or uniformly mixed.

Results

Compared with Zn sulfates, Zn oxide fertilisers had very low water solubility and slow dissolution rates, because of a higher dissolution pH. This did not translate to a diminished ability to supply Zn to plants when both sources of Zn were mixed through the soil, but there was significantly less fertiliser recovery from Zn oxides than from Zn sulfates when the fertiliser was banded near the seed.

Conclusions

All sources will be equally effective if uniformly mixed through the profile. In no-till systems where fertiliser is banded near seed, Zn sulfate is superior to Zn oxide.