Organoarsenicals such as methylarsenical methylarsenate (MAs(V)) and aromatic arsenicals including roxarsone (4-hydroxy-3-nitrophenylarsenate or Rox(V)) have been extensively used as an herbicide and growth enhancers in animal husbandry, respectively. They undergo environmental degradation to more toxic inorganic arsenite (As(III)) that contaminates crops and drinking water. We previously identified a bacterial gene (arsI) responsible for aerobic MAs(III) demethylation. The gene product, ArsI, is a Fe(II)-dependent extradiol dioxygenase that cleaves the carbon-arsenic (C-As) bond in MAs(III) and trivalent aromatic arsenicals. The objective of this study was to elucidate the ArsI mechanism. Using isothermal titration calorimetry, we determined the dissociation constants (Kd) and ligand-to-protein stoichiometries (N) of ArsI for Fe(II), MAs(III) and aromatic phenyl arsenite. Using a combination of methods including chemical modification, site-directed mutagenesis, and fluorescent spectroscopy, we demonstrated that amino acid residues predicted to participate in Fe(II)-binding (His5-His62-Glu115) and substrate binding (Cys96-Cys97) are all involved in catalysis. Finally, the products of Rox(III) degradation were identified as As(III) and 4-hydroxy-2-nitrophenol, demonstrating that ArsI is a dioxygenase that incorporates one oxygen atom from dioxygen into the carbon and the other to the arsenic to catalyze the cleavage of the C-As bond. These results augment our understanding of the mechanism of this novel C-As lyase.

Eukaryotic translation initiation factor eIF4E1 (eIF4E1) plays a pivotal role in the control of cap-dependent translation initiation, occurs in P- bodies and is important for the formation of stress granules (SG). Human cells encompass two other non-canonical translation initiation factors capable of cap binding although with a lower affinity for the cap: eIF4E2 and eIF4E3. Here, I investigated the ability of individual eIF4E family members and their variants to localize to SGs and P-bodies in stress-free, arsenite and heat shock conditions. Under all tested conditions, both eIF4E1 and eIF4E2 proteins and all their variants localized to P-bodies unlike eIF4E3 protein variants. Under both arsenite and heat stress conditions all tested variants of eIF4E1 and the variant eIF4E3-A localized to SGs albeit with different abilities. Protein eIF4E2 and all its investigated variants localized specifically to a major part of heat stress-induced stress granules. Further analysis showed that approximately 75% of heat stress-induced stress granules contain all three eIF4Es, while in 25% of them eIF4E2 is missing. Large ribosomal subunit protein L22 was found specifically enriched in arsenite induced SGs. Heat stress-induced re- localization of several proteins typical for P-bodies such as eIF4E2, DCP-1, AGO-2...

Förorenad mark är ett problem som kan hota ekologiska värden samt skada människors hälsa. Det finns flera åtgärder för föroreningar både ex-situ och in-situ. På senare tid har användningen av nollvärt järn som in-situ åtgärd studerats, men har inte använts tidigare i Sverige. Denna studie innefattar ett pilotförök i fält där en in-situ injektering av nollvärt järn i nanostorlek (nZVI) i Hjältevad, Eksjö kommun. Där bedrev dåvarande Televerket en impregneringsanläggning för telefonstolpar där de använde koppar, krom och arsenik (CCA) som efter ett läckage i en lagringstank förorenade grundvattnet på platsen.  Innan denna studie visade skak och kolonnförsök att tillsats av nZVI till jordprover från Hjältevad minskade koncentrationen av både total arsenik och arsenit med över 90 %. Syftet med pilotförsöket var att undersöka hur långt nZVI skulle spridas i akvifären och om arsenikhalten och grundvattenkemin påverkades på samma sätt av nZVI injekteringen som i labbförsöken.  Dynamiska grundvattenprovtagningar utfördes innan behandlingen för att kunna karakterisera grundvattnet i försöksytan. I ett inledande försök uppskattades spridningen av en 5 g/L nZVI slurry med tillsatt 0,75 g/L polyakrylsyra som injekterades med direct push teknologi. Spridningen av en nZVI injektering undersöktes genom mätning av elektrisk konduktivitet samt genom att observera färgen på grundvattnet. En spridning av nZVI mellan 0,6 och 1,5 m observerades och influensradien ökade med djupet i försöksytan samt visar på större vertikal utbredning vid grundare djup. Preferentiellt flöde kunde ses och överlappande injekteringspunkter behövs för en heltäckande utbredning. Pilotförsöket omfattade fem injekteringspunkter som borrades med 2 m avstånd vid källtermen. Sex nya grundvattenrör installerades uppströms och nedströms injektionspunkterna. Vattenprover togs under 4 månader där grundvattenkemiska parametrar som pH, EH, total As, trevärd As, och Fe koncentrationer mättes. Arsenikhalten låg runt 1700 µg/L innan behandlingen och har inte minskat 4 månader efter injektering av nZVI. I grundvattenrören närmast injekteringspunkterna fanns en tendens till ökad totalhalt och andel arsenit. Möjliga orsaker diskuteras och en hypotes är att lägre temperatur i akvifären jämfört med labbförsök förklarar lägre reaktivitet för nZVI. / Soil contamination poses ecological risks and enhances risks for human exposure to chemical in many areas of the world. Various ex-situ and in-situ soil remediation strategies have therefore been developed. The in-situ use of zero-valent iron sorbents is a relatively new strategy that has not yet been used in the field in Sweden. The current study involves a pilot study of an in-situ inejction of nano-scale zerovalent iron (nZVI) in a contaminated site in Hjältevad, Eksjö municipality. The former Swedish State-owned telephone compmnay operated a plant where telegraph poles were impregnated with chromated copper arsenate (CCA) that leaked into the groundwater following a tank rupture.   Batch and column tests done prior to the current study in the lab showed that addition of nZVI to sediment sampled from the aquifer reduced the dissolved concentrations of total arsenic as well as arsenite with more than 90 %. The aim of this pilot test was therefore firstly to investigate to investigate whether the same reduction could be obtained in the field.  Dynamic groundwater sampling was carried out before the pilot test to characterize hydrology and chemistry of the aquifer. A 5 g/L nZVI slurry was injected using direct push technology initially in one injection well to investigate spread of nZVI by measuring electrical conductivity and observing the color of the groundwater. The measured electrical conductivity and color observation showed a spread of nZVI between 0.6 and 1.5 m. The radius of influence increased with depth in the test zone and shows greater vertical spread at shallower depths. Preferential flow could be seen and overlapping injection points are thus needed for a complete coverage of the source zone. Five injection well were then drilled into the source zone at a distance of 2 m. New groundwater wells were installed downstream from the source zone where samples were taken over a period of 4 months in which chemical parameters such as pH, EH, total As, trivalent As, and Fe concentrations were measured. Arsenic concentrations were 1700 µg/L before the treatment and this concentration as well as other measured groundwater paramters did not decrease significantly during the 4 months monitoring period. In the groundwater wells closest to the injection points, the concentration of total and trivalent arsenic increased significantly. Possibly reasons are discussed and it is hypothesized that the lower aquifer temperature in the field comapred to the lab experiments may explain the lower nZVI reactivity.