Sirpa Peräniemi

Metallothioneins 2 and 3 contribute to the metal-adapted phenotype but are not directly linked to Zn accumulation in the metal hyperaccumulator, Thlaspi caerulescens

To study the role of metallothioneins (MTs) in Zn accumulation, the expression of TcMT2a, TcMT2b, and TcMT3 was analysed in three accessions and 15 F3 families of two inter-accession crosses of the Cd/Zn hyperaccumulator Thlaspi caerulescens, with different degrees of Zn accumulation. The highest expression levels were found in the shoots of a superior metal-accumulating calamine accession from St Laurent le Minier, with >10-fold TcMT3 expression compared with another calamine accession and a non-metallicolous accession. Moreover, F3 sibling lines from the inter-accession crosses that harboured the MT2a or MT3 allele from St Laurent le Minier had higher expression levels. However, there was no co-segregation of TcMT2a or TcMT3 expression and Zn accumulation. To examine the functions of TcMTs in plants, TcMT2a and TcMT3 were ectopically expressed in Arabidopsis. The transformant lines had reduced root length in control medium but not at high metal concentrations, suggesting that the ectopically expressed proteins interfered with the physiological availability of essential metals under limited supply. The Arabidopsis transformant lines did not show increased tolerance to Cd, Cu, or Zn, nor increased Cd or Zn accumulation. Immunohistochemical analysis indicated that in roots, MT2 protein is localized in the epidermis and root hairs of both T. caerulescens and Arabidopsis thaliana. The results suggest that TcMT2a, TcMT2b, and TcMT3 are not primarily involved in Zn accumulation as such. However, the elevated expression levels in the metallicolous accessions suggests that they do contribute to the metal-adapted phenotype, possibly through improving Cu homeostasis at high Zn and Cd body burdens. Alternatively, they might function as hypostatic enhancers of Zn or Cd tolerance.

Focal cerebral ischemia in rats alters APP processing and expression of Aβ peptide degrading enzymes in the thalamus

We have previously demonstrated aggregation of amyloid precursor protein (APP) and beta-amyloid (Abeta) to dense plaque-like deposits in the thalamus of rats subjected to transient middle cerebral artery occlusion (MCAO). Here, we investigated the underlying molecular effects of MCAO on APP processing and expression profiles of Abeta degrading enzymes in the cortex adjacent to the infarct (penumbra) and ipsilateral thalamus 2, 7 and 30 days after ischemic insult. Enhanced beta-amyloidogenic processing of APP and altered insulin degrading enzyme and neprilysin expression were observed in the thalamus, but not the penumbral cortex, 7 and 30 days after MCAO coinciding with increased calcium levels and beta-secretase (BACE) activity. Consecutively, increased BACE activity associated with depletion of BACE trafficking protein GGA3, suggesting a post-translational stabilization of BACE. These results demonstrate that focal cerebral ischemia leads to complex pathogenic events in the thalamus long after the initial insult.

Isolation of Zn-responsive genes from two accessions of the hyperaccumulator plant Thlaspi caerulescens

Several populations with different metal tolerance, uptake and root-to-shoot transport are known for the metal hyperaccumulator plant Thlaspi caerulescens. In this study, genes differentially expressed under various Zn exposures were identified from the shoots of two T. caerulescens accessions (calaminous and non-calaminous) using fluorescent differential display RT-PCR. cDNA fragments from 16 Zn-responsive genes, including those encoding metallothionein (MT) type 2 and type 3, MRP-like transporter, pectin methylesterase (PME) and Ole e 1-like gene as well as several unknown genes, were eventually isolated. The full-length MT2 and MT3 sequences differ from those previously isolated from other Thlaspi accessions, possibly representing new alleles or isoforms. Besides the differential expression in Zn exposures, the gene expression was dependent on the accession. Thlaspi homologues of ClpP protease and MRP transporter were induced at high Zn concentrations. MT2 and PME were expressed at higher levels in the calaminous accession. The MTs and MRP transporter expressed in transgenic yeasts were capable of conferring Cu and Cd tolerance, whereas the Ole e 1-like gene enhanced toxicity to these metals. The MTs increased yeast intracellular Cd content. As no significant differences were found between Arabidopsis and Thlaspi MTs, they apparently do not differ in their capacity to bind metals. However, the higher levels of MT2 in the calaminous accession may contribute to the Zn-adapted phenotype.

Zirconium-loaded activated charcoal as an adsorbent for arsenic, selenium and mercury

The adsorption of arsenic, selenium and mercury from aqueous solutions onto zirconium-loaded activated charcoal was studied as a function of adsorption time, amount of adsorbent, pH, concentration of adsorbates, sample volume and the oxidation states of the adsorbates. The cross-interference of the analytes was also investigated. Loaded filters were measured by energy-dispersive X-ray fluorescence spectrometry (EDXRF) and the amount of the unadsorbed analytes were determined by vapour generation atomic absorption spectrometry (VGAAS).

Environmental impact of mining activities on the surface water quality in Tibet: Gyama valley.

Nearly 20years of industrial scale metal mining operations in Tibet have caused an impact on the regions surface water quality. However, no information with respect to the pollution has been provided to the public. The aim of this work was to evaluate the chemical quality of the stream water and to assess the present and future potential risks of acid mine drainage to the regional and downstream environments. This study, based on data collected in 2006, 2007 and 2008 in the Gyama valley, using the Environmental Risk Index (I(ER)) documents that elevated concentrations of Cu, Pb, Zn, Mn, Fe and Al in the surface water and streambed at the upper/middle part of the valley pose a considerably high risk to the local environment. In contrast, the risk level at the stream source area is zero and only minor risk at the lower reaches. The iron and copper contamination of the upper/middle part of the river appears to be both natural and accelerated by the mining activities. The level of dissolved contaminants in the water decreases within short distance downstream due to precipitation and sorption to the streambed and strong dilution by a tributary stream and eventually by the Lhasa River. A high content of heavy metals in the stream sediments as well as in a number of tailings with gangue and material from the ore processing, poses a great potential threat to the downstream water users. Environmental changes such as global warming or increased mining activity may increase the mobility of these pools of heavy metals.

Determination of trace elements in heavy oil samples by graphite furnace and cold vapour atomic absorption spectrometry after acid digestion

A procedure was developed to determine the concentrations of trace elements (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb and V) in heavy oil samples. The wet digestion (HNO3-H2SO4) method developed for the sample preparation was both repeatable and quantitative and allowed element determinations by graphite furnace atomic absorption spectrometry (GFAAS) or cold vapour atomic absorption spectrometry (CVAAS) from the same sample solution. The detection limits were low, and low concentrations of analyte could be determined accurately. V and Ni were determined also by inductively coupled plasma atomic emission spectrometry (ICP-AES), in samples merely diluted with xylene (1:10), the 1CP-AES results were in good agreement with the GFAAS results.

Optimized arsenic, selenium and mercury determinations in aqueous solutions by energy dispersive x-ray fluorescence after preconcentration onto zirconium-loaded activated charcoal

Abstract Adsorption onto zirconium-loaded activated charcoal (ZrC ∗) allows the efficient collection of arsenic, selenium and mercury from aqueous solutions in a thin layer, and at the same time removes or minimizes elemental interferences. When this preconcentration method is combined with energy dispersive x-ray fluorescence, the resulting analysis is accurate and precise. The method is simple and rapid and almost free from contamination because only a small amount of sodium hydroxide or nitric acid is required for pH adjustment, in addition to ZrC ∗ adsorbent. Spectral interferences from other elements are negligible because the ZrC ∗ adsorbent is highly selective. The procedure is also suitable for the determination of organometallic compounds of arsenic, selenium and mercury.

Separation of microgram quantities of Cr(III) and Cr(VI) in aqueous solutions and determination by energy dispersive X-ray fluorescence spectrometry

Abstract A procedure was developed for the separation and independent determination of microgram quantities of Cr(VI) and Cr(III) in aqueous solution. Cr(VI) was adsorbed onto zirconium-loaded activated charcoal (ZrC ∗), and Cr(III) was collected by Fe(OH) 3 precipitation and the precipitate bound to activated charcoal. Chromium in the activated charcoal was measured by energy dispersive X-ray fluorescence spectrometry. The detection limits for both chromium species were better than 0.05 mg/l. The procedure was tested with the Ground Water and Waste Water Pollution Control Check Standard WP-15 spiked with Cr(VI) and several samples from the plating industry.

Advanced material and approach for metal ions removal from aqueous solutions

A Novel approach to remove metals from aqueous solutions has been developed. The method is based on a resin free, solid, non-toxic, microcrystalline bisphosphonate material, which has very low solubility in water (59 mg/l to ion free Milli-Q water and 13 mg/l to 3.5% NaCl solution). The material has been produced almost quantitatively on a 1 kg scale (it has been prepared also on a pilot scale, ca. 7 kg) and tested successfully for its ability to collect metal cations from different sources, such as ground water and mining process waters. Not only was this material highly efficient at collecting several metal ions out of solution it also proved to be regenerable and reusable over a number of adsorption/desorption, which is crucial for environmental friendliness. This material has several advantages compared to the currently used approaches, such as no need for any precipitation step.

Environmental fate and bioavailability of wood preservatives in freshwater sediments near an old sawmill site

Impacts of an old contaminated sawmill site located in Eastern Finland were studied, with emphasis on transportation and bioaccumulation of wood preservatives in the surrounding water system. To assess the transportation of chlorophenols and chromated copper arsenate (CCA) from the sawmill to the nearby lake, the concentrations of these compounds in selected sediment samples were analyzed. To assess the contribution of a pulp mill further upstream, the concentration of extractable organic halogens (EOX) was analyzed. Bioaccumulation of wood preservatives from sediments was examined using Lumbriculus variegatus as test organism. In sediments collected from the sawmill area, concentrations of chlorophenols, arsenic, chromium and copper were high. In the surrounding area the concentrations of these compounds were slightly elevated at some sampling points but were mostly within the natural range of variation. Thus, it can be concluded that transportation of wood preservatives from the sawmill area to its surroundings is fairly low. However, 60 microg/l of arsenic and 50 microg/l of copper were found in water taken from a brook that runs through a landfill area of the sawmill to the nearby river, and the concentration of arsenic in the surface sediment at one sampling point in the lake was slightly elevated. The total amount of organohalogens in sediment was higher in the river and the lake than in the sawmill area. Of all the wood preservatives studied, only arsenic was found to bioaccumulate in present conditions, reaching a tissue concentration of 362 microg/g dw in organisms exposed for 28 days to sediment from the brook. High concentration of arsenic in oligochaeta tissue was related to high concentration of arsenic in the pore water.