Overall Retention of Methyl Stereochemistry during B12-Dependent Radical SAM Methyl Transfer in Fosfomycin Biosynthesis.

Methylcobalamin-dependent radical S-adenosylmethionine (SAM) enzymes methylate non-nucleophilic atoms in a range of substrates. The mechanism of the methyl transfer from cobalt to the receiving atom is still mostly unresolved. Here we determine the stereochemical course of this process at the methyl group during the biosynthesis of the clinically used antibiotic fosfomycin. In vitro reaction of the methyltransferase Fom3 using SAM labeled with 1H, 2H, and 3H in a stereochemically defined manner, followed by chemoenzymatic conversion of the Fom3 product to acetate and subsequent stereochemical analysis, shows that the overall reaction occurs with retention of configuration. This outcome is consistent with a double-inversion process, first in the SN2 reaction of cob(I)alamin with SAM to form methylcobalamin and again in a radical transfer of the methyl group from methylcobalamin to the substrate. The methods developed during this study allow high-yield in situ generation of labeled SAM and recombinant expression and purification of the malate synthase needed for chiral methyl analysis. These methods facilitate the broader use of in vitro chiral methyl analysis techniques to investigate the mechanisms of other novel enzymes.

Determination of Characteristic vs Anomalous 135Cs/137Cs Isotopic Ratios in Radioactively Contaminated Environmental Samples.

A contamination with the ubiquitous radioactive fission product 137Cs cannot be assigned per se to its source. We used environmental samples with varying contamination levels from various parts of the world to establish their characteristic 135Cs/137Cs isotope ratios and thereby allow their distinction. The samples included biological materials from Chernobyl and Fukushima, historic ashed human lung tissue from the 1960s from Austria, and trinitite from the Trinity Test Site, USA. After chemical separation and gas reaction shifts inside a triple quadrupole ICP mass spectrometer, characteristic 135Cs/137Cs isotope signatures (all as per March 11, 2011) were obtained for Fukushima- (∼0.35) and Chernobyl-derived (∼0.50) contaminations, in agreement with the literature for these contamination sources. Both signatures clearly distinguish from the characteristic high ratio (1.9 ± 0.2) for nuclear-weapon-produced radiocesium found in human lung tissue. Trinitite samples exhibited an unexpected, anomalous pattern by displaying a low (<0.4) and nonuniform 135Cs/137Cs ratio. This exemplifies a 137Cs-rich fractionation of the plume in a nuclear explosion, where 137Cs is a predominant species in the fireball. The onset of 135Cs was delayed because of the longer half-life of its parent nuclide 135Xe, causing a spatial separation of gaseous 135Xe from condensed 137Cs, which is the reason for the atypical 135Cs/137Cs fractionation in the fallout at the test site.

Uranium concentrations in sediment pore waters of Lake Neusiedl, Austria.

The goal of the present investigation was to measure 234U/238U activity ratios in pore waters of Lake Neusiedl, Austria, in order to learn more about uranium in groundwaters of the Lake Neusiedl/Seewinkel region. Pore waters of waterlogged sediments (at 1m depth) in the littoral zone of Lake Neusiedl were analyzed. The pore water samples were collected in the National Park Lake Neusiedl/Seewinkel from pristine sites that were not influenced by neighboring fertilized fields or vineyards. Uranium isotopes were extracted from 1.5L of sediment pore water and measured by α-particle spectrometry. Uranium concentrations were found to be unexpectedly high (up to 853µgL-1) especially in pore waters of salt-rich locations. 234U/238U activity ratios were between 0.91 and 1.09 for all pore water samples, irrespective of their origin from the east or west littoral zones of the lake. Uranium and mineral salts concentrations were strongly correlated. 222Rn concentrations were low (between 22 and 42BqL-1). The results provide insight into the high degree of mobility of U(VI) in sedimentary environments, in the presence of migrating Na2SO4-type saline waters.

Sphagnum-dominated bog systems are highly effective yet variable sources of bio-available iron to marine waters.

Iron is a micronutrient of particular interest as low levels of iron limit primary production of phytoplankton and carbon fluxes in extended regions of the world's oceans. Sphagnum-peatland runoff is extraordinarily rich in dissolved humic-bound iron. Given that several of the world's largest wetlands are Sphagnum-dominated peatlands, this ecosystem type may serve as one of the major sources of iron to the ocean. Here, we studied five near-coastal creeks in North Scotland using freshwater/seawater mixing experiments of natural creek water and synthetic seawater based on a (59)Fe radiotracer technique combined with isotopic characterization of dissolved organic carbon by Accelerator Mass Spectrometry. Three of the creeks meander through healthy Sphagnum-dominated peat bogs and the two others through modified peatlands which have been subject to artificial drainage for centuries. The results revealed that, at the time of sampling (August 16-24, 2014), the creeks that run through modified peatlands delivered 11-15µg iron per liter creek water to seawater, whereas the creeks that run through intact peatlands delivered 350-470µg iron per liter creek water to seawater. To find out whether this humic-bound iron is bio-available to marine algae, we performed algal growth tests using the unicellular flagellated marine prymnesiophyte Diacronema lutheri and the unicellular marine green alga Chlorella salina, respectively. In both cases, the riverine humic material provided a highly bio-available source of iron to the marine algae. These results add a new item to the list of ecosystem services of Sphagnum-peatlands.

River-derived humic substances as iron chelators in seawater.

The speciation of iron(III) in oxic seawater is dominated by its hydrolysis and sedimentation of insoluble iron(III)-oxyhydroxide. As a consequence, many oceanic areas have very low iron levels in surface seawater which leads to iron deficiency since phytoplankton require iron as a micronutrient in order to grow. Fortunately, iron solubility is not truly as low as Fe(III) solubility measurements in inorganic seawater would suggest, since oceanic waters contain organic molecules which tend to bind the iron and keep it in solution. Various iron-binding organic ligands which combine to stabilize dissolved iron have been detected and thoroughly investigated in recent years. However, the role of iron-binding ligands from terrestrial sources remains poorly constrained. Blackwater rivers supply large amounts of natural organic material (NOM) to the ocean. This NOM (which consists mainly of vascular plant-derived humic substances) is able to greatly enhance iron bioavailability in estuaries and coastal regions, however, breakdown processes lead to a rapid decrease of river-derived NOM concentrations with increasing distance from land. It has therefore been argued that the influence of river-derived NOM on iron biogeochemistry in offshore seawater does not seem to be significant. Here we used a standard method based on 59Fe as a radiotracer to study the solubility of Fe(III)-oxyhydroxide in seawater in the presence of riverine NOM. We aimed to address the question how effective is freshwater NOM as an iron chelator under open ocean conditions where the concentration of land-derived organic material is about 3 orders of magnitude smaller than in coastal regions, and does this iron chelating ability vary between NOM from different sources and between different size fractions of the river-borne NOM. Our results show that the investigated NOM fractions were able to substantially enhance Fe(III)-oxyhydroxide solubility in seawater at concentrations of the NOM ≥ 5 µg L- 1. Terrigenous NOM concentrations ≥ 5 µg L- 1 are in no way unusual in open ocean surface waters especially of the Arctic and the North Atlantic Oceans. River-derived humic substances could therefore play a greater role as iron carriers in the ocean than previously thought.

Extraction of natural radionuclides from aqueous solutions by novel maltolate-based task-specific ionic liquids.

Two novel maltol-based ionic liquids, namely [A336][Mal] and [C101][Mal], were synthesized as potential extracting agents for radionuclides from water. These two room temperature task-specific ionic liquids could be easily prepared by anion metathesis starting from commercially available materials. The isolated compounds were characterized by standard analytical methods. Their application as extraction agent for Unat., 234Th, 210Pb, 210Bi, 210Po and 226Ra was elucidated by liquid-liquid extraction and scintillation counting. Uranium was totally extracted by both ionic liquids over a broad pH range (2-8), while the other radionuclides were removed with differing efficacies depending on the respective pH value.

A review on 129I analysis in air.

A review of literature focused on (129)I determination in air is provided. (129)I analysis in the environment represents a vital tool for tracing transport mechanisms, distribution pathways, safety assessment and its application as environmental tracer. To achieve that, specific chemical extraction methods and high sensitivity analytical techniques have been developed. This paper is intended to give an overview about the sample collection, extraction and distribution of (129)I in the air. Sensitivity of available measurement techniques for the determination of (129)I is compared. The article also provides the summary of current worldwide distribution of (129)I in air and respective radiation exposure of man.

Investigation of the isotopic ratio 129I/I in petrified wood.

In fossil specimens, measurements of the natural isotopic ratio (129)I/I may provide a method to estimate the age of sample. The motivation for measuring the isotopic composition ((129)I/I) of petrified wood samples collected from Austria was to check this feasibility. Alkaline fusion together with anion exchange was used to extract iodine from the sample. Typical sample size for this study was 10-90 g. An atomic ratio as low as 10(-14) was determined using accelerator mass spectrometry (AMS). Uranium concentrations measured by instrumental neutron activation analysis (INAA) and α-spectrometry were found to be less than 3 mg kg(-1), therefore the contribution from fissiogenic (129)I was small and an estimation of ages was based on the decrease of the initial ratio (due to decay of the cosmogenic (129)I in a closed system) after subtraction of the fissiogenic (129)I. The value of the prenuclear ratio is crucial for the use of the (129)I system for dating purposes in the terrestrial environment. From the preanthropogenic (initial) ratio of 1.5 × 10(-12) of the hydrosphere and the results of the present study for the samples from Altenburg (1.05 × 10(-12)) and Fuerwald (6.16 × 10(-13)), respective ages of 8 ± 2.2 and 20.2 ± 2.2 million years were derived. Since samples were collected from a stratum deposited in the Upper Oligocene/Ergerien period (~25-30 million years ago), it can be concluded that these isotopic ratios do not show ages but an elapsed time since fossil wood was isolated from mineral rich water. Paleontological investigation shows that samples from Altenburg had mixed characteristics of old and modern Tertiary plants, thus an origin from a younger stratum re-sedimented with Oligocene cannot be excluded. However, the sample from Drasenhofen reflects that the (129)I/I system might not always be suitable for the dating of petrified wood sample due to fixation of anthropogenic (129)I into surface fractures.

Iodine isotopes (127I and 129I) in aerosols at high altitude Alp stations.

Concentrations of gases and particulate matter have been proven to be affected by meteorological and geographical variables from urban locations to high mountain clean air sites. Following our previous research in Vienna, we summarize here new findings about concentration levels of iodine isotopes in aerosols collected at two Alpine meteorological stations, Sonnblick (Austria) and Zugspitze (Germany) during 2001. The present study mainly focuses on the effect of altitude on the anthropogenic concentration of (129)I and on the isotopic ratio (129)I/(127)I. Iodine was separated from matrix elements by using either an anion exchange method or solvent extraction, and was analyzed by ICP-MS and AMS. Over the altitude change from Vienna to Zugspitze and Sonnblick (202 m to 2962 m and 3106 m above sea level), stable iodine level decreased from an average of 0.94 ng m(-3) to 0.52 ng m(-3) and 0.62 ng m(-3), respectively. Similarly, (129)I concentrations at both Alpine stations were about 1 order of magnitude lower (10(4) atoms m(-3)) than values obtained for Vienna (10(5) atoms m(-3)) and reveal a strong vertical concentration gradient of (129)I. A high degree of variability is observed, which is due to wide variation in the origin of air masses. Furthermore, air trajectory analysis demonstrates the importance of large scale air transport mostly from southeast Europe for influencing Sonnblick whereas influence from northwest Europe is strong at Zugspitze. In contrast to (129)I, a higher concentration of (7)Be was found at higher altitude stations compared to Vienna which probably results from its production in the upper atmosphere.

Retrospective measurements of airborne 129iodine in Austria.

The knowledge about the distribution of anthropogenic (129)I is crucial for a successful establishment of transport mechanisms, fate and behaviour in the environment. In present study, the historical record of dry deposition of (129)I in Austria over four decades back to the 1960s is reconstructed. The (129)I/(127)I isotopic ratio of the order of 10(-9)-10(-7) in airborne particles revealed a prominent anthropogenic (129)I signature. The time profile of airborne (129)I follows directly the pattern of the gaseous emissions from European reprocessing plants. Furthermore, temporal variations of (129)I were traced monthly over two years. The potential risks of internal exposure to (129)I are associated with both inhalation and ingestion. Since dose via inhalation was found insignificant, the thyroid equivalent dose from the internal exposure of (129)I using a value of 10(-8) for the isotopic ratio (129)I/(127)I in the thyroid and ICRP reference man was calculated. The corresponding thyroid cancer risk factor of 10(-11) for an adult from life-time exposure is one order of magnitude higher than for a 1-year old child. Due to low radiation toxicity of (129)I the annual dose is 8 × 10(4) times lower than the dose limit of the National Research Council, USA which is 0.04 mSv y(-1) to whole body or any organ for a combined beta and photon emitting radionuclide.

Monitoring of radionuclides in soil and bone samples from Austria.

The activity concentrations of anthropogenic (9°Sr, ¹³7Cs) and natural (²³8U, ²³²Th, 4°K, ²¹°Pb) radionuclides were determined in eight soil profiles from three different regions in Austria (Styria, Carinthia and Salzburg). A direct correlation between the activity concentration of 9°Sr and ¹³7Cs in soil samples and site altitude was found. 9°Sr and ²¹°Pb activity concentrations were also determined in bone ash of deer hunted in these regions. Additional bone samples were collected all over Austria. Totally 39 deer bones were investigated for this work, and some values were adopted from our earlier publications to give an overview of samples collected from sites covering a broad range of altitudes. The bone and soil samples were collected in the time period of 2001-2009. The 9°Sr values in deer bones are directly proportional to the values in the respective soil samples and also to the age of the animals. For the 9°Sr and ²¹°Pb determinations in bone samples first Pb was separated on a Dowex column, then Sr was purified using Sr·Spec® resin. In soil samples an additional hydroxide precipitation was employed to eliminate interfering iron. For the first time also the 3M Empore® Sr Rad disk method was successfully applied to bone samples. With this method the chemical procedure can be shortened by more than a factor of 2. The 9°Sr and ²¹°Pb fractions were measured by liquid scintillation counting, while the chemical yields were determined by ICP-MS. The activity concentrations of 4°K, ²³8U, ²³²Th and ¹³7Cs in soil samples were evaluated using gamma-ray spectrometry. The investigation was part of the PhD work of the first author.

Natural radionuclides in Austrian mineral water and their sequential measurement by fast methods.

Ten samples of Austrian mineral water were investigated with regard to the natural radionuclides (228)Ra, (226)Ra, (210)Pb, (210)Po, (238)U and (234)U. The radium isotopes as well as (210)Pb were measured by liquid scintillation counting (LSC) after separation on a membrane loaded with element-selective particles (Empore Radium Disks) and (210)Po was determined by alpha-spectroscopy after spontaneous deposition onto a copper planchette. Uranium was determined by ICP-MS as well as by alpha-spectroscopy after ion separation and microprecipitation with NdF(3). From the measured activity concentrations the committed effective doses for adults and babies were calculated and compared to the total indicative dose of 0.1 mSv/a given in the EC Drinking Water Directive as a maximum dose. The dominant portion of the committed effective dose was due to the radium isotopes; the dose from (228)Ra in most samples clearly exceeded the dose from (226)Ra.

Determination of strontium-90 in deer bones by liquid scintillation spectrometry after separation on Sr-specific ion exchange columns.

The activity concentration of (90)Sr was determined in several deer bones from Austria. Strontium specific ion exchange columns with 4',4''(5'')-di-t-butylcyclohexane-18-crown-6 from Eichrom Industries, Inc. were used for separation. The yield of the chemical procedure was quantified with AAS. Directly after column separation, the solution containing (90)Sr was mixed with the scintillation cocktail HiSafe III and measured by liquid scintillation counting. Prevention of (210)Pb contamination and reusability of the separation columns was investigated as well as the activity distribution within the bones. Results were compared with pre-Chernobyl measurements in Austria; a correlation between activity concentration of (90)Sr and site altitude was found.

Aerosols: unexpected disequilibrium phenomena between airborne radio activities of lead-210 and its progenies bismuth-210 and polonium-210.

For the first time, concentrations of the long lived radon progenies (210)Pb, (210)Bi and (210)Po were measured in the mine atmosphere of the so called "healing gallery" in Badgastein, Austria, a region famous for its radioactive springs. These investigations were performed in order to study the radioactive equilibrium between the (210)Pb-(210)Bi and the (210)Pb-(210)Po pairs so as to gain more information about the aerosol-forming processes in the mine. The particle size distribution of the aerosols was determined under different ventilation conditions. Six-stage and eight-stage cascade impactors with working ranges from 0.15 to 5 micro m and from 0.063 to 8 micro m, respectively, were used to collect the mine aerosols. These samples were analysed in the laboratory and measured by liquid scintillation spectrometry. The most surprising results were found under full ventilation, when the total activity concentrations of (210)Pb, (210)Bi and (210)Po were 4.6, 2.0 and 16.5 mBq/m(3), respectively. In this case (210)Po/(210)Pb activity ratios ranged between 1.8+/-0.3 and 4.3+/-0.3. These unexpected results were confirmed by the eight-stage impactor samples. For the smallest particles, between 0.062 and 0.125 micro m, an even higher value of 7.5 was observed. As outside sources could be excluded, such (210)Po enrichments must occur during the aerosol-forming process itself inside the mine.