Arsenobetaine amide: a novel arsenic species detected in several mushroom species.

The total arsenic mass fraction as well as the arsenic speciation were studied in four different mushroom species with inductively coupled plasma mass spectrometry and high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry, respectively. Arsenic mass fractions detected in the mushrooms were covering a range from 0.3 to 22 mg As kg-1 dry mass. For the arsenic speciation, species like arsenobetaine, inorganic arsenic, or dimethylarsinic acid were found, which are commonly detected in mushrooms, but it was also proven that the recently discovered novel compound homoarsenocholine is present in Amanita muscaria and Ramaria sanguinea. Moreover, a previously unidentified arsenic species was isolated from Ramaria sanguinea and identified as trimethylarsonioacetamide, or in short: arsenobetaine amide. This new arsenical was synthesized and verified by spiking experiments to be present in all investigated mushroom samples. Arsenobetaine amide could be an important intermediate to further elucidate the biotransformation pathways of arsenic in the environment.

Formation of Colloidal In(As,P) Quantum Dots Active in the Short-Wave Infrared, Promoting Growth through Temperature Ramps.

Colloidal InAs quantum dots (QDs) are widely studied as a printable optoelectronic material for short-wave infrared (SWIR) that is not restricted by regulations on hazardous substances. Such applications, however, require synthetic procedures that yield QDs with adjustable sizes at the end of the reaction. Here, we show that such one-size-one-batch protocols can be realized through temperature profiles that involve a rapid transition from a lower injection temperature to a higher reaction temperature. By expediting the transition to the reaction temperature and reducing the overall synthesis concentration, we can tune QD sizes from 4.5 to 10 nm, the latter corresponding to a band gap transition at 1600 nm. We argue that the temperature ramps provide a more distinct separation between nucleation at low temperature and growth at high temperature such that larger QDs are obtained by minimizing the nucleation time. The synthetic procedures introduced here will strongly promote the development of a SWIR optoelectronic technology based on InAs QDs, while the use of temperature profiles to steer a colloidal synthesis can find applications well beyond the specific case of InAs QDs.

Genetic Susceptibility to Neurotoxicity Related to Prenatal Inorganic Arsenic Exposure in Young Spanish Children.

We explored the influence of child and maternal single nucleotide polymorphisms (SNPs) in genes related to neurological function and arsenic metabolism (i.e., ABCA1, ABCB1, PON1, CYP3A, BDNF, GSTP1, MT2A, and APOE as well as AS3MT) on the association between prenatal arsenic (As) exposure and methylation efficiency and neuropsychological development in 4-5-year-old children. Participants were 549 mother-child pairs from the INMA (Environment and Childhood) Spanish Project. We measured inorganic arsenic (iAs) and the metabolites monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) in urine samples collected during pregnancy. Neuropsychological development was assessed at the age of 4-5 years using the McCarthy Scales of Children's Abilities (MSCA). Several SNPs were determined in maternal and child DNA; AS3MT and APOE haplotypes were inferred. The median ∑As (sum of iAs, DMA, and MMA) was 7.08 µg/g creatinine. Statistically significant interactions for children's APOE haplotype were observed. Specifically, ε4-carrier children had consistently lower MSCA scores in several scales with increasing ∑As and MMA concentrations. These results provide evidence regarding the neurotoxic effects of early life exposure to As, observing that the APOE ε4 allele could make children more vulnerable to this exposure.

Influence of genetic polymorphisms on arsenic methylation efficiency during pregnancy: Evidence from a Spanish birth cohort.

BACKGROUND: Inorganic arsenic (iAs) is a widespread toxic metalloid. It is well-known that iAs metabolism and its toxicity are mediated by polymorphisms in AS3MT and other genes. However, studies during pregnancy are scarce. We aimed to examine the role of genetic polymorphisms in AS3MT, GSTO2, N6AMT1, MTHFR, MTR, FTCD, CBS, and FOLH1 in iAs methylation efficiency during pregnancy. METHODS: The study included 541 pregnant participants from the INMA (Environment and Childhood) Spanish cohort. Using high-performance liquid chromatography coupled to inductively coupled plasma-tandem mass, we measured arsenic (iAs and the metabolites monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA)) in urine samples collected during the first trimester. iAs methylation efficiency was determined based on relative concentrations of the As metabolites in urine (%MMA, %DMA, and %iAs). Thirty-two single nucleotide polymorphisms (SNPs) in nine genes were determined in maternal DNA; AS3MT haplotypes were inferred. We assessed the association between genotypes/haplotypes and maternal As methylation efficiency using multivariate linear regression models. RESULTS: The median %MMA and %DMA were 5.3 %, and 89 %, respectively. Ancestral alleles of AS3MT SNPs (rs3740393, rs3740390, rs11191453, and rs11191454) were significantly associated with higher %MMA, %iAs, and lower %DMA. Pregnant participants with zero copies of the GGCTTCAC AS3MT haplotype presented a higher %MMA. Statistically significant associations were also found for the FOLH1 SNP rs202676 (ß 0.89 95%CI: 0.24, 1.55 for carriers of the G allele vs. the A allele). CONCLUSIONS: Our study shows that ancestral alleles in AS3MT polymorphisms were associated with lower As methylation efficiency in early pregnancy and suggests that FOLH1 also plays a role in As methylation efficiency. These results support the hypothesis that As metabolism is multigenic, being a key element for identifying susceptible populations.

Cadmium in the hyperaccumulating mushroom Thelephora penicillata: Intracellular speciation and isotopic composition.

Thelephora penicillata is an ectomycorrhizal mushroom that can accumulate extraordinarily high concentrations of Cd, As, Cu, and Zn in its fruit-bodies. To better understand its element accumulation ability, we compared the element concentrations in T. penicillata with 10 distinct ectomycorrhizal mushroom species growing at the same site (Karlina Pila, Czech Republic). On average, T. penicillata accumulated 330, 2130, 26, and 4 times more Cd, As, Cu, and Zn, respectively, than other mushrooms. Size-exclusion chromatography and an electrophoretic analysis of T. penicillata cell extracts indicate that intracellular Cd may be present mainly in >1 kDa, presumably compartmentalized, Cd species, and partially binding with 6-kDa cysteinyl-containing peptide(s) resembling metallothioneins. The cadmium isotopic composition of mushroom fruit-bodies, soil digests, and soil extracts was investigated by thermal ionization mass spectrometry (TIMS) with double spike correction. The isotopic composition (δ114/110Cd) of ectomycorrhizal mushrooms from Karlina Pila varied in a wide range of -0.37 to +0.14 ‰. However, remarkably low δ114/110Cd values were observed in the majority of the investigated mushrooms when compared to the relatively homogeneous Cd isotopic composition of bulk soil (δ114/110Cd = +0.09 ‰) and the comparatively heavy isotopic composition of soil extracts (mean δ114/110Cd values of +0.11 ± 0.01 ‰ and +0.22 ± 0.01 ‰, depending on the extraction method). The isotopic composition of Cd hyperaccumulated in T. penicillata essentially matched the mycoavailable soil Cd fraction. However, most isotopic data indicates isotopic fractionation at the soil/fruit-body interface, which could be of environmental significance.

Quantitative mapping of mercury and selenium in mushroom fruit bodies with laser ablation-inductively coupled plasma-mass spectrometry.

This work describes the development of a novel method for quantitative mapping of Hg and Se in mushroom fruit body tissues with laser ablation coupled to inductively coupled plasma-mass spectrometry (LA-ICP-MS). Different parameters of the protocol for preparation of the standards used for quantification via external calibration were assessed, e.g., the dissolution temperature of gelatin standards and the addition of chitosan and L-cysteine as additives to the gelatin-based calibration droplets to better match the sample matrix. While chitosan was not suited for this purpose, the presence of L-cysteine considerably improved the figures of merit of the calibration, leading to limits of detection of 0.006 and 0.3 µg g-1 for Hg and Se, respectively, at a pixel size of 20 × 20 µm. Further, an in-house reference material, ideally suited for the validation of the method for application to mushroom samples, was successfully prepared from a paste of Boletus edulis. The newly developed method was used to investigate the distribution of Hg and Se in tissue sections of five porcini mushroom individuals of three different species (Boletus edulis, Boletus aereus, and Boletus pinophilus) and one sample of a parasol mushroom (Macrolepiota procera). For one sample, additional areas were ablated at higher spatial resolution, with a laser spot size down to 5 µm, which allows a detailed investigation of the spatial distribution of Hg and Se in mushrooms.

A new mushroom hyperaccumulator: Cadmium and arsenic in the ectomycorrhizal basidiomycete Thelephora penicillata.

Fruit-bodies of six Thelephora species (Fungi, Basidiomycota, Thelephoraceae) were analyzed for their trace element concentrations. In Thelephora penicillata, extremely high concentrations of Cd and As were found, followed by highly elevated concentrations of Cu and Zn. The highest accumulation ability was found for Cd with a mean concentration of 1.17 ± 0.37 g kg-1 (dry mass) in fruit-bodies collected from 20 unpolluted sites; the mean As concentration was 0.878 ± 0.242 g kg-1. Furthermore, striking accumulation of Se (923 ± 28 mg kg-1) was found in one sample of T. vialis and elevated concentrations of S were detected in T. palmata (19.6 ± 5.9 g kg-1). The analyzed Thelephora species were sequenced and, based on the Maximum Likelihood phylogenetic analysis (ITS rDNA) of the genus, possible other Thelephora (hyper)accumulators were predicted on the basis of their phylogenetic relationship with the discovered (hyper)accumulators. The striking ability of T. penicillata to accumulate simultaneously Cd, As, Cu, and Zn has no parallel in the Fungal Kingdom and raises the question of a biological importance of metal(loid) hyperaccumulation in mushrooms.

Arsenic speciation analysis in honey bees for environmental monitoring.

Arsenic can be toxic to living organisms, depending not only on the concentration, but also its chemical form. The aim of this study was to determine arsenic concentrations and perform arsenic speciation analysis for the first time in honeybees, to evaluate their potential as biomonitors. Highest arsenic concentrations were determined in the vicinity of coal fired thermal power plants (367 µg kg-1), followed by an urban region (213 µg kg-1), with much lower concentrations in an industrial city (28.8 µg kg-1) and rural areas (41 µg kg-1). Until now, honey bees have never been used to study different arsenic species in the environment. For this reason, four extraction procedures were tested: water, hot water at 90 °C, 20% methanol, and 1% formic acid. Water at 90 °C was able to extract more than 90% of the total arsenic from honey bee samples. Inorganic arsenic (the sum of arsenite and arsenate) accounted for 95% of arsenic species in bees from three locations, except the industrial city, where it represented only 80% of arsenic species, while 15% was present as DMA.

Prenatal arsenic exposure, arsenic methylation efficiency, and neuropsychological development among preschool children in a Spanish birth cohort.

BACKGROUND: Prenatal arsenic (As) exposure could negatively affect child neuropsychological development, but the current evidence is inconclusive. OBJECTIVES: To explore the relationship between prenatal urinary total As (TAs) concentrations, the As species and the methylation efficiency, and child neuropsychological development in a Spanish birth cohort. We also studied the effect modification produced by sex and several nutrients and elements. MATERIALS AND METHODS: Study subjects were 807 mother-child pairs participating in the INMA (Childhood and Environment) Project. Urinary TAs and its metabolites, monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), inorganic As (iAs) and arsenobetaine were measured in the first trimester of pregnancy. Methylation efficiency was determined through the percentages of the metabolites and using principal component analysis. Children's neuropsychological development was assessed at the age of 4-5 years using the McCarthy Scales of Children's Abilities (MSCA). Multivariable linear regression models were built to assess the association between TAs, the As species and the maternal methylation efficiency, and the neuropsychological scores. We explored effect modification by sex, iron status, maternal nutrients status (serum manganese and selenium, and urinary zinc), and maternal vitamins intake (folate, and vitamins B12 and B6). RESULTS: The geometric mean (95%CI) of ∑As (sum of DMA, MMA and iAs) was 7.78 (7.41, 8.17) µg/g creatinine. MMA concentrations were inversely associated with the scores for the general, verbal, quantitative, memory, executive function and working memory scales (i.e. ß [CI95%] = -1.37 [-2.33, -0.41] for the general scale). An inverse association between %MMA and the memory scores was found. Children whose mothers had lower manganese, zinc and ferritin concentrations obtained lower scores on several MSCA scales with decreasing As methylation efficiency. DISCUSSION: An inverse association was observed between MMA concentrations and children's neuropsychological development. Maternal levels of manganese, zinc and ferritin affected the association between As methylation efficiency and MSCA scores.

Trace Element Accumulation in Two Turtle Species, Malaclemys terrapin and Chelydra serpentina, in New Jersey, USA.

Trace elements in aquatic environments pose a risk to biological communities; this study investigates the total concentrations of arsenic (As), silver (Ag), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), mercury (Hg), nickel (Ni), selenium (Se), lead (Pb), and zinc (Zn) within muscle, carapace, liver, and adipose tissues of diamondback terrapins and common snapping turtles in New Jersey. The effects of tissue type, sex, size, and location upon trace element accumulation were studied. The data obtained indicates that within diamondback terrapins and common snapping turtles, trace element accumulations displayed a significant difference among tissue types and sex (p < 0.005). The data indicates that Ag, Cd, Cu, and Hg can accumulate within the liver of diamondback terrapin. Se was found to accumulate in the livers of both diamondback terrapin and common snapping turtles. The highest mean concentrations of Co, Cr, Ni, and Pb were found in the carapace of both turtle species. Sex was found to have an impact on As, Hg, and Zn accumulation within different tissue types of diamondback terrapins. Diamondback terrapin males were found to have higher concentrations of As within the carapace. Diamondback terrapin females possessed higher concentrations of Hg in muscle tissues and Hg and Zn in the carapace. Turtle size and collection location land type and land cover did not display any correlation with trace element bioaccumulation for either species.

Parallel and comparative non-targeted metabolomic speciation analysis of metalloids and their non-metal analogues by HPLC-ICPMS/MS in mushrooms.

With the introduction of tandem mass spectrometry to inductively coupled plasma mass spectrometry (ICPMS/MS), the potential for non-targeted elemental metabolomic analysis has been expanded to many non-metals of pivotal biological importance. Arsenic and selenium are trace elements that share chemical similarity with the non-metals phosphorus and sulfur, respectively, and this similarity can be exploited to gain more insight into the incompletely understood biological significance of these metalloids and the evolution of their biochemical pathways. As a proof of concept, we show the applicability of HPLC-ICPMS/MS for non-targeted and parallel speciation analysis of arsenic, selenium, phosphorus, and sulfur in mushrooms-metabolically diverse organisms. Incredibly contrasting levels of diversity were found in the metabolomic profiles of the four investigated elements among the various species along with sharp discrepancies among related elements (e.g. phosphorous vs. arsenic) in certain mushroom species. The present work shows that ICPMS/MS offers a new dimension in non-targeted metabolomic analysis and enables a unique comparative approach in investigating and tracking the biochemistry of related elements in moderately complex organisms.

Is the water disinfection by-product dichloroacetic acid biosynthesized in the edible mushroom Russula nigricans?

We report the first halogen speciation analysis study by high performance liquid chromatography coupled with inductively coupled plasma tandem mass spectrometry (HPLC-ICPMS/MS) in the fruiting bodies of various mushroom species. Non-targeted speciation analysis revealed the occurrence of dichloroacetic acid (DCAA) in the edible mushroom Russula nigricans. Multiple samples of this mushroom (n = 5) collected from different geographic non-industrial regions in two different countries confirmed the consistent presence of this species at a relatively narrow concentration range (23-37 mg kg-1), whereas no other chlorinated acetic acid (e.g. chloroacetic acid and trichloroacetic acid) was detected. Neither DCAA nor any other chlorinated acetic acid were detected in any of the other mushroom species investigated in the present study, including seven different mushroom species of the same genus Russula, even though all mushrooms were collected from the same non-industrial geographic regions. Together with the previously reported biological activities of DCAA, these findings collectively suggest biosynthesis of this compound as an explanation for its dominant presence in R. nigricans, and constitute the first example of the dominant natural occurrence of this compound over other chlorinated acetic acids in a living organism. This may warrant a change in our view of the occurrence of dichloroacetic acid in nature, where primarily considered as a pollutant arising from water disinfection.

Urinary arsenic species and methylation efficiency during pregnancy: Concentrations and associated factors in Spanish pregnant women.

BACKGROUND: Arsenic (As) is considered to be toxic for humans, the main routes of exposure being through drinking water and the diet. Once ingested, inorganic arsenic can be methylated sequentially to monomethyl and dimethyl arsenicals. Several factors can affect both As exposure and methylation efficiency. OBJECTIVES: To describe the urinary concentrations of the different As species and evaluate the methylation efficiency during pregnancy, as well as their associated factors in a birth cohort of pregnant Spanish women. METHODS: Participants in this cross-sectional study were 1017 pregnant women from two areas of Spain who had taken part in the INMA (Environment and Childhood) project (2003-2008). Total As (organic and inorganic compounds) and its main metabolites (monomethylarsonic acid, [MMA], dimethylarsinic acid, [DMA], inorganic As [iAs]) and arsenobetaine [AB]) were measured in urine samples collected during the first trimester. Sociodemographic and dietary information was collected through questionnaires. Multivariate linear regression models were used to explore the association between As species concentrations and covariates. Arsenic methylation efficiency was determined through the percentages of the metabolites and using As methylation phenotypes, obtained from principal component analysis. RESULTS: Median urine concentrations were 33.0, 21.6, 6.5, 0.35 and 0.33 µg/g creatinine for total As, AB, DMA, MMA and iAs, respectively. Daily consumption of rice and seafood during the first trimester of pregnancy were positively associated with the concentration of As species (i.e., ß [CI95%] = 0.36 [0.09, 0.64] for rice and iAs, and 1.06 [0.68, 1.44] for seafood and AB). TAs, AB and iAs concentrations, and DMA and MMA concentrations were associated with legume and vegetable consumption, respectively. The medians of the percentage of As metabolites were 89.7 for %DMA, 5.1 for %MMA and 4.7 for %iAs. Non-smoker women and those with higher body mass index presented a higher methylation efficiency (denoted by a higher %DMA and lower %MMA). DISCUSSION: Certain dietary, lifestyle, and environmental factors were observed to have an influence on both As species concentrations and methylation efficiency in our population. Further birth cohort studies in low exposure areas are necessary to improve knowledge about arsenic exposure, especially to inorganic forms, and its potential health impact during childhood.

Arsenocholine-O-sulfate: A novel compound as major arsenic species in the parasitic mushroom Tolypocladium ophioglossoides.

The As concentrations, along with 34 other elements, and the As speciation were investigated in wild-grown samples of the parasitic mushroom Tolypocladium ophioglossoides with inductively coupled plasma mass spectrometry (ICPMS) and high performance liquid chromatography coupled to ICPMS. The As concentrations were 0.070-3.44 mg kg-1 dry mass. More remarkable was the As speciation, where up to 56% of the extracted As were found to be an unknown As species, which was marginally retained under anion- and also cation-exchange conditions. After testing several different chromatographic settings, the compound was finally isolated and identified as 2-(sulfoxyethyl) trimethylarsonium ion (in short: arsenocholine-O-sulfate) with high resolution mass spectrometry. The compound was synthesized and further quantified in all investigated samples via ion-pair chromatography coupled to ICPMS. In addition to the high abundance of arsenocholine-O-sulfate in T. ophioglossoides, small amounts of this As species were also detected in one sample of the host mushroom, Elaphomyces asperulus. In a sample of another parasitic mushroom, Ophiocordyceps sinensis, arsenocholine-O-sulfate could not be detected, but the main species was another unknown compound that was oxidized to inorganic As(V) with hydrogen peroxide. This is the first discovery of arsenocholine-O-sulfate in nature. It is possible that it is present in many other organisms, at least in low concentrations, and just has not been detected there yet because of its unusual chromatographic behavior. The existence of arsenocholine-O-sulfate brings up questions again about the biotransformation pathways of As in the environment and the specific behavior of fungi.

Metal Ion Promiscuity and Structure of 2,3-Dihydroxybenzoic Acid Decarboxylase of Aspergillus oryzae.

Broad substrate tolerance and excellent regioselectivity, as well as independence from sensitive cofactors have established benzoic acid decarboxylases from microbial sources as efficient biocatalysts. Robustness under process conditions makes them particularly attractive for preparative-scale applications. The divalent metal-dependent enzymes are capable of catalyzing the reversible non-oxidative (de)carboxylation of a variety of electron-rich (hetero)aromatic substrates analogously to the chemical Kolbe-Schmitt reaction. Elemental mass spectrometry supported by crystal structure elucidation and quantum chemical calculations verified the presence of a catalytically relevant Mg2+ complexed in the active site of 2,3-dihydroxybenoic acid decarboxylase from Aspergillus oryzae (2,3-DHBD_Ao). This unique example with respect to the nature of the metal is in contrast to mechanistically related decarboxylases, which generally have Zn2+ or Mn2+ as the catalytically active metal.

Is arsenic responsible for the toxicity of the hyperaccumulating mushroom Sarcosphaera coronaria?

The Violet Crown Cup, Sarcosphaera coronaria, is a rather inconspicuous mushroom, but with an interesting and unresolved mystery. In earlier days, the mushroom was considered edible, but several poisonings were reported in the early 20th century. The reason for the seemingly sporadic toxicity of S. coronaria is still unknown. One possible explanation is arsenic, since Crown Cups can take up high amounts of this element. We investigated the arsenic concentration and arsenic speciation in S. coronaria with inductively coupled plasma mass spectrometry (ICPMS) and HPLC coupled to ICPMS and found up to incredible 0.9% As (dry mass). Most of it was present as methylarsonic acid (MA), a less toxic form of this element. However, low concentrations of the highly toxic methylarsonous acid [MA (III)] were also detected. The amounts were too low to pose an acute risk for consumers, but the concentration of MA (III) significantly increased during simulated gastric digestion. We could not unambiguously identify arsenic as the toxic constituent of S. coronaria, but we demonstrated that the extremely toxic MA (III) can be formed under certain circumstances, which should be carefully investigated in future.

Arsenic species in mushrooms, with a focus on analytical methods for their determination - A critical review.

Since the discovery of the "fish arsenic" arsenobetaine in a mushroom, arsenic species have been studied in all kinds of mushrooms, with very diverse results, for more than 20 years. Depending on the fungal species, inorganic arsenic, methylarsonic acid, dimethylarsinic acid, arsenobetaine and a couple of other arsenic-containing compounds have been detected in all kinds of concentrations in natural samples. A rule to predict the arsenic speciation of a new mushroom is yet to be found. Some research has been devoted to the question about the origin of the different arsenic species in mushrooms. It is still not clear whether mushrooms are able to transform arsenic by themselves or if they just take it up from the environment. For the determination of the arsenic species in mushrooms, high performance liquid chromatography coupled to inductively coupled plasma mass spectrometry is used most of the times, but a few alternative approaches exist. This review will give an overview of these different techniques and critically discuss them. The arsenic species that have been found in mushrooms so far will be summarized, and the work that has been devoted to unraveling the origins of these compounds will be discussed in depth. The interesting phenomenon of arsenic hyperaccumulation by mushrooms will be reviewed as well.

Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients.

Polcalcins are important respiratory panallergens, whose IgE-binding capacity depends on the presence of calcium. Since specific immunotherapy is not yet available for the treatment of polcalcin-sensitized patients, we aimed to develop a molecule for efficient and safe immunotherapy. We generated a hypoallergenic variant of the grass pollen polcalcin Phl p 7 by introducing specific point mutations into the allergen's calcium-binding regions. We thereby followed a mutation strategy that had previously resulted in a hypoallergenic mutant of a calcium-binding food allergen, the major fish allergen parvalbumin. Dot blot assays performed with sera from Phl p 7-sensitized patients showed a drastically reduced IgE reactivity of the Phl p 7 mutant in comparison to wildtype Phl p 7, and basophil activation assays indicated a significantly reduced allergenic activity. Rabbit IgG directed against mutant rPhl p 7 blocked patients' IgE binding to wildtype Phl p 7, indicating the mutant's potential applicability for immunotherapy. Mass spectrometry and circular dichroism experiments showed that the mutant had lost the calcium-binding capacity, but still represented a folded protein. In silico analyses revealed that the hypoallergenicity might be due to fewer negative charges on the molecule's surface and an increased molecular flexibility. We thus generated a hypoallergenic Phl p 7 variant that could be used for immunotherapy of polcalcin-sensitized individuals.

Simultaneous selenium and sulfur speciation analysis in cultivated Pleurotus pulmonarius mushroom.

Selenium (Se) and sulfur (S) speciation analysis in edible and medicinal Se enriched P. pulmonarius extracts was performed. Mycelium, colonized substrate, and fruiting bodies at different harvesting times were analyzed using ion-pairing reversed-phase chromatography coupled to an ICPMS/MS detector. Extraction efficiencies in enzymatically digested and aqueous extracts were between 45.3 and 109% for Se, depending on the sample type. Selenomethionine (Se-Met) was found to be the major Se-compound, together with a number of unknown Se-species. Cystine (Cys2), methionine (Met), and sulfate were also detected and quantified in all samples. Most of the Se-Met (84.0%) and Met (75.8%) were found to be in free form in the fruiting body, in contrast with the mycelium where 53.4% of Se-Met and 80.5% of Met is incorporated into proteins.

Speciation analysis of elements accumulated in Cystoderma carcharias from clean and smelter-polluted sites.

Collections of Cystoderma carcharias sporocarps were sampled from clean and smelter-polluted sites and analyzed for Ag, As, Cd, Cu, Pb, Se, and Zn contents. Concentrations of all elements were significantly higher in samples from the smelter-polluted area. Except for As and Pb, all elements were effectively accumulated in the sporocarps at both clean and polluted sites. With the highest concentration of 604 mg Cd kg-1, C. carcharias can be considered as Cd hyperaccumulator. As revealed by HPLC-ICPQQQMS analysis, the As species in sporocarps from clean and polluted areas involved besides the major arsenobetaine a variety of known and unknown arsenicals; the occurrence of dimethylarsinoylacetate and trimethylarsoniopropionate is reported for the first time for gilled fungi (Agaricales). Size-exclusion chromatography of C. carcharias extracts supported by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and RP-HPLC data indicated that detoxification of intracellular Cd and Cu may largely rely on metallothioneins (MT) or MT-like peptides, not phytochelatins.