Protolytic cleavage of Hg-C bonds induced by 1-methyl-1,3-dihydro-2H-benzimidazole-2-selone: synthesis and structural characterization of mercury complexes.

Multinuclear ((1)H, (77)Se, and (199)Hg) NMR spectroscopy demonstrates that 1-methyl-1,3-dihydro-2H-benzimidazole-2-selone, H(sebenzim(Me)), a structural analogue of the selenoamino acid, selenoneine, binds rapidly and reversibly to the mercury centers of HgX2 (X = Cl, Br, I), while X-ray diffraction studies provide evidence for the existence of adducts of composition [H(sebenzim(Me))]xHgX2 (X = Cl, x = 2, 3, 4; X = I, x = 2) in the solid state. H(sebenzim(Me)) also reacts with methylmercury halides, but the reaction is accompanied by elimination of methane resulting from protolytic cleavage of the Hg-C bond, an observation that is of relevance to the report that selenoneine demethylates CysHgMe, thereby providing a mechanism for mercury detoxification. Interestingly, the structures of [H(sebenzim(Me))]xHgX2 exhibit a variety of different hydrogen bonding patterns resulting from the ability of the N-H groups to form hydrogen bonds with chlorine, iodine, and selenium.

Exposición al metilmercurio en la población general; toxicocinética; diferencias según el sexo, factores nutricionales y genéticos / Methylmercury exposure in the general population; toxicokinetics; differences by gender, nutritional and genetic factors

El mercurio es un tóxico ambiental que causa numerosos efectos adversos en la salud humana y en los ecosistemas naturales. Los factores que determinan la aparición de efectos adversos y su severidad son entre otros: la forma química del mercurio (elemental, inorgánico, orgánico), la dosis, la edad, la duración de la exposición, la vía de exposición y los factores ambientales, nutricionales y genéticos. En el ciclo acuático del mercurio, una vez que se ha depositado, se transforma en metilmercurio por la acción de determinadas bacterias sulfato reductoras y se bioacumula en los organismos acuáticos incorporándose a la cadena trófica de alimentos. El contenido de metilmercurio es mayor en las especies depredadoras de mayor tamaño y que viven más años como el emperador, pez espada, tiburón, atún o marlín. El metilmercurio se halla unido a las proteínas del pescado por lo que no se elimina mediante la limpieza ni el cocinado del mismo. El feto en desarrollo y los niños pequeños son los más vulnerables a los efectos neurotóxicos del metilmercurio procedente de la ingesta de pescado contaminado. El metilmercurio se absorbe en el tracto gastrointestinal y atraviesa la barrera hematoencefálica y la placenta. Algunos componentes de la dieta como los ácidos grasos poliinsaturados, el selenio, la fibra, los compuestos tiol, algunos fitoquímicos y otros nutrientes pueden modificar la bioaccesibilidad del mercurio y su toxicidad. Además de los factores ambientales, los factores genéticos pueden influir en la toxicidad del mercurio y explicar parte de la vulnerabilidad individual (AU)

Mercury is an environmental toxicant that causes numerous adverse effects on human health and natural ecosystems. The factors that determine the existance of adverse effects, as well as their severity are, among others: the chemical form of mercury (elemental, inorganic, organic), dosis, age, period of exposure, pathways of exposure and environmental, nutritional and genetic factors. In the aquatic cycle of mercury, once it has been deposited, it is transformed into methylmercury due to the action of certain sulphate-reducing bacteria, which bioaccumulates in the aquatic organisms and moves into the food chain. The methylmercury content of large, long-lived fish such as swordfish, shark, tuna or marlin, is higher. Methylmercury binds to protein in fish and is therefore not eliminated by cleaning or cooking the fish. Fetuses and small children are more vulnerable to the neurotoxic effects of methylmercury from the consumption of contaminated fish. Methylmercury is absorbed in the gastrointestinal tract and crosses the blood-brain barrier and the placenta. The intake of certain dietary components such as polyunsaturated fatty acids, selenium, fiber, thiol compounds, certain phytochemicals and other nutrients can modify methylmercury bioaccesibility and its toxicity. Apart from environmental factors, genetic factors can influence mercury toxicity and explain part of the individual vulnerability (AU)

Aeration prevents methyl mercury production in dental wastewater.

Although research has demonstrated that Hg is methylated in the reducing conditions of the dental clinic wastewater collection system, studies are inconclusive as to whether further methylation occurs in the aeration basin of activated sludge wastewater treatment plant (WWTP) which typically treats this waste. Given the high levels of methyl Hg reported in dental wastewater (DWW), it is important to determine whether additional methylation occurs once it enters the WWTP. To achieve this objective, we incubated DWW under conditions designed to mimic the oxidized conditions of the activated sludge aeration basin in a WWTP. Duplicate bioreactors were charged with raw DWW collected from a 12-chair dental clinic and incubated both with and without aeration. Aeration was continued for 15 days, consistent with the typical mean cell residence time (MCRT) necessary for both heterotrophic carbon oxidation (typically 5-6 days) and nitrification (typically 12-15 days), thus ensuring that incubation time exceeded those for most conceivable MCRTs used in the activated sludge process. Results demonstrate a rapid increase in pH concomitant with an increase in dissolved oxygen (DO) to near saturation in the aerated reactor. The non-aerated reactor remained low or at zero DO due to low surface reaeration coupled with the high levels of organic matter. The rate of mercury methylation increased in the unaearated reactors rapidly upon incubation, reaching highest levels when DO was at the lowest levels during the experiment. In great contrast, methyl mercury levels were much lower and net mercury methylation does not appear to occur at any significant rate under aeration. These results imply that although some mercury methylation may occur in the sewer collection system (or anaerobic digesters), net methylation is unlikely to occur in the aeration basin in activated sludge WWTPs, and thus methyl Hg influent levels from DWW represent an upper bound on effluent levels.

Nitrate controls methyl mercury production in a streambed bioreactor.

Organic carbon bioreactors provide low-cost, passive treatment of a variety of environmental contaminants but can have undesirable side effects in some cases. This study examines the production of methyl mercury (MeHg) in a streambed bioreactor consisting of 40 m³ of wood chips and designed to treat nitrate (NO3) in an agricultural drainage ditch in southern Ontario (Avon site). The reactor provides 30 to 100% removal of NO3-N concentrations of 0.6 to 4.4 mg L(-1), but sulfate (SO4(2-)) reducing conditions develop when NO3 removal is complete. Sulfate reducing conditions are known to stimulation the production of MeHg in natural wetlands. Over one seasonal cycle, effluent MeHg ranged from 0.01 to 0.76 ng L(-1) and total Hg ranged from 1.3 to 3.4 ng L(-1). During all sampling events when reducing conditions were only sufficient to promote NO3(-) reduction (or denitrification) ( = 5, late fall 2009, winter 2010), MeHg concentrations decreased in the reactor and it was a net sink for MeHg (mean flux of -5.1 µg m(-2) yr(-1)). During all sampling events when SO4(2-) reducing conditions were present ( = 6, early fall 2009, spring 2010), MeHg concentrations increased in the reactor and it was a strong source of MeHg to the stream (mean flux of 15.2 µg m(-2) yr(-1)). Total Hg was consistently removed in the reactor (10 of 11 sampling events) and was correlated to the total suspended sediment load ( r² = 0.69), which was removed in the reactor by physical filtration. This study shows that organic carbon bioreactors can be a strong source of MeHg production when SO4(2-) reducing conditions develop; however, maintaining NO3-N concentrations > 0.5 mg L suppresses the production of MeHg.

A theoretical study of abiotic methylation reactions of gaseous elemental mercury by halogen-containing molecules.

Methylation reactions of gaseous elementary mercury by halogen containing molecules such as halogenomethane species CH(3)X (with X = Cl, Br, and I) and the dimethylchlorinium ion CH(3)ClCH(3)(+) were investigated at the density functional level. With CH(3)X, the reaction is predicted to be almost athermic and kinetically demanding for a thermal reaction. The reaction can proceed photochemically in the visible range; therefore sunlight may increase the reaction rate. These results compare well with the experimental data. Consecutive methylation of the CH(3)HgX products (with X = Cl, Br, and I) and subsequent formation of CH(3)HgCH(3) were also studied. These reactions are predicted to be kinetically inaccessible and thermodynamically unfavorable. With CH(3)ClCH(3)(+), the reaction is predicted to be athermic but kinetically easy. This is due to the suitability of the methyl transfer reagent. Geometrical and electronic data were systematically analyzed in order to rationalize the results.

Chemical demethylation of methylmercury by selenoamino acids.

A new chemical demethylation pathway for methylmercury under physiologically and environmentally relevant conditions is reported. The pathway involves the reaction between methylmercury and a selenoamino acid (L-selenocysteine, L-selenoglutathione, D,L-selenopenicillamine, or L-selenomethionine) via the formation of bis(methylmercuric)selenide and dimethylmercury as intermediates. The final degradation product is HgSe(s).

Synthesis, characterization and structures of methylmercury complexes with selenoamino acids.

Four new methylmercury-selenoamino acid complexes were synthesized, including methylmercury-L-selenoglutathionate, methylmercury-D,L-selenopenicillaminate, and two methylmercury-L-selenomethioninate complexes (one via a Hg-Se bonding and the other Hg-N bonding). All the complexes were characterized by NMR ((1)H, (13)C, (77)Se and (199)Hg), FT-IR and mass spectra. Their molecular structures were established by single crystal X-ray crystallography (for the Hg-N bonding methylmercury-L-selenomethioninate) and by quantum mechanical calculations using Gaussian-03 with the hybrid functional B3LYP/SDD. All four complexes were found to chemically and structurally resemble their sulfur analogues, with a slightly stronger binding affinity of Hg to Se than to S, suggesting chemical and structural mimicry might play a role in methylmercury-selenium antagonism in biological systems.

Spatial characteristics of net methylmercury production hot spots in peatlands.

Many wetlands are sources of methylmercury (MeHg) to surface waters, yet little information exists about the distribution of MeHg within wetlands. Total mercury (THg) and MeHg in peat pore waters were studied in four peatlands in spring, summer, and fall 2005. Marked spatial variability in the distribution of MeHg, and %MeHg as a proxy for net MeHg production, was observed, with highest values occurring in discrete zones. We denote these zones "MeHg hot spots", defined as an area where the pore water %MeHg exceeded the 90th percentile of the data set (n=463) or >22% of THg as MeHg. MeHg hot spots occurred near the interface between peatland and the upland watershed with few exceptions. The %MeHg in pore water was significantly less in peatland interiors compared to upland-peatland interface zones, with the significance of these differences related to the delineation of the boundary between the two areas. Although further research is necessary, our data suggest that the occurrence of MeHg hot spots is related to the transport of solutes in upland runoff to the peatland perimeter and not to the accumulation of MeHg in this zone as a result of transport from either the peatland interior or the surrounding upland watershed. These findings augment the understanding of peatland MeHg production in upland-peatland watersheds, provide guidance for more accurate quantification of MeHg pool sizes in the landscape, and a spatial framework forthe further study of mercury methylation processes in peatlands.

Investigation of mercury-containing proteins by enriched stable isotopic tracer and size-exclusion chromatography hyphenated to inductively coupled plasma-isotope dilution mass spectrometry.

In order to investigate trace mercury-containing proteins in maternal rat and their offspring, a method of enriched stable isotopic tracer (196Hg and 198Hg) combined with size-exclusion chromatography (SEC) coupled to inductively coupled plasma-isotope dilution mass spectrometry (ICP-IDMS) was developed. Prior to the analysis, 196Hg- and 198Hg-enriched methylmercury was administrated to the pregnant rats. Then the mercury-containing proteins in serum and brain cytosol of the dam and pup rats were separated by size-exclusion columns and the mercury was detected by ICP-MS. The ICP-MS spectrogram of the tracing samples showed significantly elevated 196Hg and 198Hg isotopic signals compared with the natural ones, indicating that the detection sensitivity could be increased by the tracer method. The contents of mercury in chromatographic fractions of the dam and pup rat brain cytosol were quantitatively estimated by post-column reverse ID-ICP-MS. The quantitative speciation differences of mercury in brain cytosol between the dam and pup rats were observed, indicating that such studies could be useful for toxicological estimation. Additionally, the isotopic ratio measurement of 198Hg/202Hg in the tracing samples could be used to identify the artifact mercury species caused in the analytical procedure. The study demonstrates that the tracer method combined with high-performance liquid chromatography (HPLC)-ICP-IDMS could provide reliably qualitative and quantitative information on mercury-containing proteins in organisms.

An improved method for the synthesis of mercurated dUTP. Enzymic synthesis of Hg-labelled DNA of high molecular weight suitable for use in an image based DNA sequencing strategy.

The development of high-resolution scanning-probe microscopes has reawakened interest in the possibility of sequencing large nucleic acid molecules by direct imaging. Such an approach would be facilitated by the availability of effective methods for increasing contrast by labelling specific nucleotides, and the utility of introducing mercury atoms into complete DNA molecules through the enzymic polymerisation of mercurated pyrimidine deoxynucleoside triphosphates has been re-investigated. A simplified and improved method for the synthesis of a heat- and thiol-stable, mercurated derivative of deoxyuridine triphosphate in high yield and the incorporation of this precursor into full-length copies of a single-stranded phage M13 template are described. The DNA product has been fully characterised and the quantitative and specific replacement of thymidylic acid residues by the mercurated analogue demonstrated.

Synthesis, spectroscopic and magnetic resonance studies of mercury (II) and methylmercury (II) complexes of azathioprine, a biologically active mercaptopurine derivative.

Synthetic and spectroscopic studies of the Hg(II) and MeHg(II) complexes of azathioprine (AZA), a biologically active 6-mercaptopurine derivative, were undertaken. The altered coordination behavior of AZA with respect to the parent mercaptopurine, with sulfur no longer being the primary donor atom, was confirmed. As concluded by the 1H NMR, 13C NMR, and IR spectroscopic data, Hg(II) binds to the N(9) position of deprotonated AZA, while in the MeHg(II) compound, coordination occurs through the N(3) and N(9) positions of the purine ring. The values of the coupling constants 2J (199Hg-1H), 1J(199Hg-13C) for the MeHg(II) compound further support complexation via nitrogen atoms of the purine. Elemental analyses confirmed the compounds to be Hg(AZA)2 (1) and [(MeHg)2(AZA)](NO3) (2); conductivity measurement values show that 1 is a nonelectrolyte and 2 is a 1:1 electrolyte. Furthermore, the FAB-MS of the compounds confirms direct binding of the metal to the ligand, and in the case of the MeHg(II) compound, the successive loss of one and two MeHg(II) moieties can be clearly observed.

In vivo incorporation of [14C]leucine into brain protein of mice treated with methylmercury and thiol complexes of methylmercury.

The effect of methylmercury and thiol complexes of methylmercury on inhibition of protein synthesis was evaluated. Mice were injected (i.p.) with the following treatments: methylmercuric chloride, methylmercury-glutathione, methylmercury-cysteinylglycine and control (vehicle) for 10 days. Ten animals from each group were injected with [14C]leucine 90 min prior to death. The brains were removed and the extracted protein was subjected to liquid scintillation analysis. Mice receiving the methylmercury and methylmercury-glutathione treatments exhibited significantly greater weight loss than the control while the methylmercury-cysteinylglycine treatment was not significantly different than the control. Incorporation of [14C]leucine into brain protein was significantly depressed in the methylmercury (81% of control) and the methylmercury-glutathione (79% of control) treatments. Protein synthesis in mice receiving the methylmercury-cysteinylglycine complex although not significantly different than the methylmercury treatments was only 92% of the control mice.

Preparation of CH3 203HgCl of high specific activity.

In order to have a methylmercury compound of much higher specific activity than was obtainable from commercial suppliers of the compound, a procedure for the methylation of radioactive mercuric chloride with no addition of non-radioactive mercury has been developed. Methylation is accomplished by reacting the mercuric chloride with tetramethyl tin. Extractions into and out of benzene twice produces a methylmercury compound which is finally dissolved in a dilute sodium carbonate solution suitable for biological experiments. The entire procedure to produce a batch of the compound may be accomplished within a single day.

A simple and rapid method for preparation of 203Hg-labeled methylmercury from 203HgCl2 and methylcobalamin.

A rapid and simple method for preparation of 203Hg-labeled methylmercuric chloride (CH3(203)HgCl) from 203HgCl2 and methylcobalamin is described. More than 99% of 203Hg was methylated with methylcobalaminin 0.01 N HCl during 1 h. CH3(203)HgCl formed during the reaction was rapidly and completely separated from cobalamins and unreacted 203HgCl2 by CM-Sephadex C-25 minicolumn. The low cost procedure for preparation of CH3(203)HgCl can be completed within 2 h and yields an inorganic 203Hg-free CH3(203)HgCl which is useful in methylmercury toxicology.

[Transformation reactions of special metals in organisms and in the environment. 2. Abiotic methylation reactions of mercury, especially by methyltin compounds and humic and fulvic acids]. / Transformationsreaktionen ausgewählter Metalle in Organismen und in der Umwelt. 2. Mitt. Abiotische Methylierungsreaktionen von Quecksilber, insbesondere durch Methylzinn-Verbindungen und Humin- und Fulvinsäuren.

Following a literature survey concerning abiological methylation reactions including transmethylation of mercury we present the results of transmethylation experiments. Model experiments in aqueous solution show an instantaneous methylation of mercuric chloride by mono-, di-, and trimethyltin chloride, the dimethyl compound having the highest rate of formation. The same is valid for similar experiments with soils, in which influences of the kind of soil also play a certain role. The methylation experiments of HgCl2 with humic and fulvic acids isolated from soil yielded positive results only in some few of the extracts and only with humic acid fractions.

The synthesis and enzymatic polymerization of 5-thio- and 5-methylmercurithio-pyrimidine nucleotides.

The 5-thio and 5-methylmercurithio derivatives of UTP, dUTP and dCTP have been synthesized and tested as substrates for nucleic acid polymerases. The 5-thio-nucleotides were polymerized inefficiently by both RNA polymerase and DNA polymerase I of Escherichia coli. The 5-methylmercurithio derivatives of dUTP and dCTP were, however, utilized by DNA polymerase I, an enzyme insensitive to mercurial compounds, although they were potent inhibitors of all other polymerases tested. While polymers containing the 5-thio substituent possess structural abnormalities, most likely interstrand disulfide bridges, polymers containing 5-methylmercurithio groups appear normal. The latter polynucleotides are readily separated from non-sulfated polymers by chromatography on mercuriagarose.