Toxicity of copper and zinc alone and in combination in Caenorhabditis elegans model of Huntington's disease and protective effects of rutin.

Copper (Cu) and Zinc (Zn) are required in small concentrations for metabolic functions, but are also toxic. There is a great concern about soil pollution by heavy metals, which may exposure the population to these toxicants, either by inhalation of dust or exposure to toxicants through ingestion of food derived from contaminated soils. In addition, the toxicity of metals in combination is questionable, as soil quality guidelines only assess them separately. It is well known that metal accumulation is often found in the pathologically affected regions of many neurodegenerative diseases, including Huntington's disease (HD). HD is caused by an autosomal dominantly inherited CAG trinucleotide repeat expansion in the huntingtin (HTT) gene. This results in the formation of a mutant huntingtin (mHTT) protein with an abnormally long polyglutamine (polyQ) repeat. The pathology of HD results in loss of neuronal cells, motor changes, and dementia. Rutin is a flavonoid found in various food sources, and previous studies indicate it has protective effects in HD models and acts as a metal chelator. However, further studies are needed to unravel its effects on metal dyshomeostasis and to discern the underlying mechanisms. In the present study, we investigated the toxic effects of long-term exposure to copper, zinc, and their mixture, and the relationship with the progression of neurotoxicity and neurodegeneration in a C. elegans-based HD model. Furthermore, we investigated the effects of rutin post metal exposure. Overall, we demonstrate that chronic exposure to the metals and their mixture altered body parameters, locomotion, and developmental delay, in addition to increasing polyQ protein aggregates in muscles and neurons causing neurodegeneration. We also propose that rutin has protective effects acting through mechanisms involving antioxidant and chelating properties. Altogether, our data provides new indications about the higher toxicity of metals in combination, the chelating potential of rutin in the C. elegans model of HD and possible strategies for future treatments of neurodegenerative diseases caused by the aggregation of proteins related to metals.

Effects of La2O3 nanoparticles and bulk-La2O3 on the development of Pfaffia glomerata (Spreng.) Pedersen and respective nutrient element concentration.

Nanoparticles (NPs) have been progressively applied in the last decades, which may impact the environment. Synthesis of pigments, growing, and nutrient element uptake by plants can also be affected by NPs. The influence of lanthanum oxide nanoparticles (La2O3 NPs) on growth, pigment synthesis, and nutrient element uptake by Pfaffia glomerata (Spreng.) Pedersen, a medicinal plant native in South America, was evaluated in the present study. P. glomerata plantlets were cultivated for 28 days in the absence (control) and presence of 100, 200, and 400 mg L-1 of La2O3 NPs or bulk-La2O3 (b-La2O3) at the same cultivation conditions. Root development, aerial part growth, and pigment concentration in plants were affected by b-La2O3 and La2O3 NPs, mainly by La2O3 NPs. In spite of alteration of nutrient element concentration observed for the 100 and 200 mg L-1 of La2O3 NPs or b-La2O3 treatments, Ca, Cu, Fe, K, La, Mg, Mn, Mo, P, S, and Zn determination in stems and leaves revealed drastically and similar decrease of these elements in plants cultivated in the presence of 400 mg L-1 of La2O3 NPs or b-La2O3. Element distribution (mapping) determined by using laser ablation inductively coupled plasma mass spectrometry in leaves of plants submitted to treatment with 400 mg L-1 of b-La2O3 or La2O3 NPs showed differences in the distribution of elements, indicating distinct effects of b-La2O3 and La2O3 NPs on P. glomerata. As such, this study demonstrated that La2O3 NPs may impact plant growth. However, more investigations are necessary for better understanding of the effect of La2O3 on plants, including a broader range of concentration.

The copper economy response is partially conserved in rice (Oryza sativa L.).

Copper (Cu) is an essential element for plants, especially in photosynthesis, as it is required for plastocyanin function in electron transfer reactions at thylakoid membranes. In Arabidopsis thaliana, Cu deficiency leads to the Cu economy response, in which plants prioritize Cu usage by plastocyanin in detriment of non-essential cupric proteins. In rice (Oryza sativa), however, this response has not been characterized. Rice OsHMA5 is a Cu xylem-loading transporter involved in Cu translocation from roots to shoots, as suggested by the analysis of oshma5 mutant plants. Aiming to understand how rice plants respond to Cu deficiency and how decreased Cu translocation to shoots can affect this response, we characterized the physiological and molecular responses of WT and oshma5 plants under control and Cu deficiency treatments. We found evidence that shoots of oshma5 plants are more prone to Cu deficiency compared to shoots of WT plants, as demonstrated by decreased chlorophyll and Cu concentrations, and electron transport rate. Gene expression analysis revealed that Cu high-affinity transporters OsCOPT1 and OsCOPT5, along with a set of miRNAs and three Cu/Zn superoxide dismutases are responsive to Cu deficiency in both WT and oshma5 plants, suggesting their involvement in the Cu economy response. However, Fe superoxide dismutase was not up-regulated in rice, indicating a difference compared to the A. thaliana Cu economy model. Therefore, we provide evidence for a partially conserved Cu economy response in rice, in comparison to A. thaliana.

Plants' genetic variation approach applied to zinc contamination: secondary metabolites and enzymes of the antioxidant system in Pfaffia glomerata accessions.

Zinc (Zn) is a micronutrient, but its excessive concentration can impair plant growth and development. Fertilizers, liming materials, pesticides and fungicides containing Zn have contributed to increase its concentration in agricultural soils. The aim of the present study is to evaluate the effect of Zn excess on the non-enzymatic (anthocyanin and ß-ecdysone) and enzymatic (superoxide dismutase-SOD and guaiacol peroxidase-GPX) antioxidant system of two P. glomerata accessions (JB and GD) grown in hydroponic system and soil, under short- and long-term exposure times. Three Zn levels (2, 100 and 200 µM) and two short-term exposure times (7 and 14 d) were tested in the hydroponic experiment. Three Zn levels (2, 100 and 200 mg kg-1) and two long-term exposure times (34 and 74 d) were tested in the soil experiment. The effects of Zn excess on P. glomerata accessions depended on the growth system and exposure time. Zinc excess in both tested growth systems resulted in significant change in the tissue oxidative process (MDA concentration) in both accessions, as well as broadened the antioxidant system response, which was based on antioxidant enzymes (SOD and GPX) and secondary metabolites (anthocyanins and ß-ecdysone). The highest anthocyanin concentration was observed in accession JB, which was grown in hydroponics, but tissue anthocyanin concentration increased in both accessions, regardless of growth medium and exposure time. The ß-ecdysone concentration in the roots increased in both accessions, but accession GD was more responsive to Zn excess. There was significant physiological variation in P.glomerata accessions in response to Zn excess.

Investigative Analysis of Lanthanum Oxide Nanoparticles on Elements in Bone of Wistar Rats After 30 Days of Repeated Oral Administration.

Accumulation of lanthanum oxide nanoparticles (La2O3NPs) in the femur bone of rats after 30 days of oral administration was explored. Also, the influence of La2O3NPs on macro and trace elements in the rats' femur bone was assessed. Inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma optical spectrometry (ICP OES) were used for total element determination in the bone after decomposition while laser ablation-ICP-MS (LA-ICP-MS) was used to investigate element distribution (bio-imaging) in the bone. Some differences in element concentrations in the bone between the rats treated with La2O3NPs at 1.0 mg kg-1 (T1), 10.0 mg kg-1 (T2), and 100 mg kg-1 (T3) body weight (bw) and the control rats (CTR) were observed. More differences were observed in the bone of rat treated with 10.0 mg kg-1 La2O3NPs bw. However, the highest change observed was for Mg, which concentration ranged from 5230 ± 12 µg kg-1 for the CTR group to 4130 ± 138 µg kg-1 for the T3 group. Minor changes were observed for Ba, Ca, Cr, Cu, Fe, Mg, Na, Pb, Sr, and Zn between CTR and animals treated with La2O3NPs at the different levels of concentration. It was possible to observe from LA-ICP-MS analysis that La2O3NPs were accumulated only on the surface of the bone, not deeper than about 5 µm. LA-ICP-MS allowed also to investigate the distribution of La and the other elements in a cross section of the femur bone head, where higher amounts of the elements are present at the external part of the bone. Therefore, it was demonstrated that La2O3NPs are incorporated on the surface of the bone and it has a small influence on some of the other elements evaluated.

Arsenic speciation analysis in rice milk using LC-ICP-MS.

The consumption of rice milk has increased, mainly by individuals intolerant to lactose or allergic to cow milk. However, rice milk contains As. In this sense, the concentration of As in rice milk should be controlled. In the present study it is proposed a methodology for determination of As(III), dimethylarsenic (DMA), monomethylarsenic (MMA) and As(V) species in rice milk using LC-ICP-MS. The main features of the methodology are fast analysis, easy and simple sample preparation, where the sample is 3-fold diluted in the mobile phase and then filtered. The four arsenic species investigated were detected in the analysed samples, being As(V) the main species. The limit of quantification of the method ranges from 0.25 to 0.43 µg L-1 As. The analyte recovery ranged from 81 to 116% for samples spiked to 1.00 µg L-1 or 5.00 µg L-1 As and the relative standard deviation was better than 5%.

In vitro stability of arsenic trioxide-liposome encapsulates for acute promyelocytic leukemia treatment.

In this work, we investigated the stability of arsenic trioxide (ATO) used in leukemia treatment, encapsulated with nanoliposome, with the aid of ultrasound treatment. Stability studies of As species were followed by liquid chromatography-inductively coupled plasma mass spectrometry (LC-ICP-MS), allowing for the detection of the conversion of low amounts of As(III) to As(V) or the formation of other As species. The influence of storage temperature and time on ATO was evaluated. Low amounts of As(III) to As(V) conversions were observed when the As encapsulated with nanoliposome was incubated at 25 °C and 40 °C. However, As(III) was stable if the solution was maintained at 5 °C, even after 90 days. No formation of other As species was observed, indicating good stability of the encapsulated ATO. Next step of the work will focus on spray drying of ATO nanoliposomes-encapsuleted with the aim of long term stability of As.

Bioimaging Metallomics.

This chapter focuses on bioimaging in metallomics, which involves metal and metalloids distribution in animal tissues. It starts with laser ablation-inductively coupled plasma-mass spectrometry followed by secondary ion mass spectrometry, synchrotron-based X-ray fluorescence, and electron microscopy, including transmission electron microscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. The basic principles of these techniques and their application for qualitative and quantitative imaging of elements are presented. Sample preparation for bioimaging is briefly discussed. The usefulness of element bioimaging is demonstrated for cells and several animal tissues, including the brain, liver, hair, eye, teeth, and bone. As such, this chapter addresses the state of the art in bioimaging metallomics.

Feasibility of Rare Earth Element Determination in Low Concentration in Crude Oil: Direct Sampling Electrothermal Vaporization-Inductively Coupled Plasma Mass Spectrometry.

A comprehensive study was developed showing the feasibility of determination of rare earth elements (REE) in low concentration in crude oil by using direct sampling electrothermal vaporization system coupled to inductively coupled plasma mass spectrometry (ETV-ICP-MS). The effect of organic modifier on the REE signal was evaluated and the use of 6 mg of citric acid allowed calibration using aqueous reference solutions (selected pyrolysis and vaporization temperatures were 700 and 2200 °C, respectively). Because of the facility of REE in forming refractory compounds inside the graphite furnace during the heating step, the use of a modifier gas (Freon R-12, 3.0 mL min-1) was necessary to allow quantitative vaporization of these elements. A flow rate of 0.40 L min-1 was selected for both bypass and carrier gases. Under optimized conditions, the influence of sample mass was evaluated, and even using a relatively high mass of crude oil (up to 18 mg), accurate results were obtained. The accuracy was evaluated by the comparison of results by ETV-ICP-MS with those obtained by ICP-MS with ultrasonic nebulizer (USN) after high-pressure microwave-assisted wet digestion (MAWD) and microwave-induced combustion (MIC) and no statistical difference was observed between the results. The limits of quantification for REE by ETV-ICP-MS were lower (0.02-0.8 ng g-1) than those obtained by USN-ICP-MS after MAWD and MIC (0.6-5.1 ng g-1). Negligible blank values and relative standard deviations lower than 12% show the feasibility of the proposed ETV-ICP-MS method for routine analysis of crude oil.

Selenium and silicon reduce cadmium uptake and mitigate cadmium toxicity in Pfaffia glomerata (Spreng.) Pedersen plants by activation antioxidant enzyme system.

Cadmium (Cd) is toxic to plants and animals, making it necessary to develop strategies that seek to reduce its introduction into food chains. Thus, the aim of this study was to investigate whether silicon (Si) and selenium (Se) reduce Cd concentrations in Pfaffia glomerata medicinal plant and attenuate the oxidative stress promoted by this metal. These plants were cultivated in hydroponics under the following treatments: control (nutrient solution), 2.5 µM Se, 2.5 mM Si, 50 µM Cd, 50 µM Cd + 2.5 µM Se, 50 µM Cd + 2.5 mM Si. After 14 days of exposure to treatments, leaves and roots were collected for the determination of dry weight of shoot and roots, Cd concentrations, chlorophyll and carotenoids content, and biochemical parameters (lipid peroxidation and guaiacol peroxidase and superoxide dismutase activities). The data were submitted to analysis of variance and means were compared with Scott-Knott test at 5% error probability. Roots of P. glomerata plants showed a significant reduction on dry weight accumulation when exposed to Cd. However, both Se and Si promoted a significant reduction of deleterious effects of Cd. The Cd concentrations in the tissues were reduced in the presence of Se or Si. Plants treated with Cd together with Se or Si presented higher pigment content than those with only Cd, thus showing a reduction in the negative effects caused by this element. In the treatments in which Se and Si were added in the growth medium together with Cd, an activation of superoxide dismutase and guaiacol peroxidase enzymes was observed in the roots and shoot, which may have contributed to lower lipid peroxidation. Thus, Se and Si reduce Cd concentrations and have potential to ameliorate Cd toxicity in P. glomerata plants, which can be used to increase productivity and quality of medicinal plants.

Selenium and mercury levels in rat liver slices co-treated with diphenyl diselenide and methylmercury.

Organoseleno-compounds have been investigated for its beneficial effects against methylmercury toxicity. In this way, diphenyl diselenide (PhSe)2 was demonstrated to decrease Hg accumulation in mice, protect against MeHg-induced mitochondrial dysfunction, and protect against the overall toxicity of this metal. In the present study we aimed to investigate if co-treatment with (PhSe)2 and MeHg could decrease accumulation of Hg in liver slices of rats. Rat liver slices were co-treated with (PhSe)2 (0.5; 5 µM) and/or MeHg (25 µM) for 30 min at 37 °C and Se and Hg levels were measured by inductively coupled plasma mass spectrometry (ICP-MS) in the slices homogenate, P1 fraction, mitochondria and incubation medium. Co-treatment with (PhSe)2 and MeHg did not significantly alter Se levels in any of the samples when compared with compounds alone. In addition, co-treatment with (PhSe)2 and MeHg did not decrease Hg levels in any of the samples tested, although, co-incubation significantly increased Hg levels in homogenate. We suggest here that (PhSe)2 could exert its previously demonstrated protective effects not by reducing MeHg levels, but forming a complex with MeHg avoiding it to bind to critical molecules in cell.

Nutritional disorder in Pfaffia glomerata by mercury excess in nutrient solution / Desordem nutricional em Pfaffia glomerata pelo excesso de mercúrio em solução nutritiva

ABSTRACT: The mineral nutritional homeostasis in response to different concentrations of Hg (0, 25 and 50μM) was evaluated in Pfaffia glomerata plant. The exposure to the highest level of Hg (50µM) caused a decreasing in shoot and root fresh weights of 15.5% and 20%, respectively. Both shoot and root Hg concentrations increased linearly with increasing external Hg concentrations. Ca concentration decreased in shoot only at 50µM Hg, whereas shoot K and Mg concentrations decreased at both 25 and 50µM Hg, when compared to the control. A significant decrease in Cu, Zn, Fe and Mn concentrations in plants exposed to Hg was observed, but most Zn, Mn, and Cu in the roots. On the other hand, P concentration increased in both root and shoot of plants exposed at 25 and 50µM Hg, whereas Na concentration increased only in the root at 25 and 50µM Hg exposure. In general, tissue nutrient concentrations in P. glomerata plantlets exposed to Hg were significantly decreased, which indicates that the Hg may cause alteration on the mineral nutritional homeostasis of this species.

RESUMO: A homeostase nutricional mineral em resposta a diferentes concentrações de Hg (0, 25 e 50μM) foram avaliadas em plantas de Pfaffia glomerata. A exposição ao mais alto nível de Hg (50µM) causou um decréscimo de 15,5% e 20%, respectivamente, na matéria fresca da parte aérea e raízes. As concentrações de Hg na parte aérea e raízes aumentaram linearmente com o aumento das concentrações de Hg. A concentração de Ca decresceu na parte aérea somente em 50µM Hg, enquanto as concentrações de K e Mg na parte aérea decresceram tanto em 25 como em 50µM Hg, quando comparado ao controle. Observou-se um significativo decréscimo nas concentrações de Cu, Zn, Fe e Mn nas plantas expostas ao Hg, mas principalmente Zn, Mn e Cu nas raízes. Por outro lado, a concentração de P aumentou em raízes e parte aérea de plantas expostas a 25 e 50µM Hg, enquanto a concentração de Na aumentou somente nas raízes em 25 e 50µM Hg. No geral, as concentrações de nutrientes nos tecidos de P. glomerata expostas ao Hg foram significativamente diminuídas, o que indica que o Hg pode causar alterações na homeostase nutricional mineral dessa espécie.

Toxic and nutrient elements in yerba mate (Ilex paraguariensis).

Toxic and nutrient elements were investigated in yerba mate (Ilex paraguariensis) from South America. Fifty-four brands of commercialised yerba mate from Argentina, Brazil, Paraguay and Uruguay were analysed for Al, Ba, Ca, Cu, Fe, K, Mg, Mn, P, Sr, and Zn, using inductively coupled plasma optical emission spectrometry (ICP-OES), and Li, Be, Ti, V, Cr, Ni, Co, As, Se, Rb, Mo, Ag, Cd, Sb, La, Ce, Pb, Bi and U using inductively coupled plasma mass spectrometry (ICP-MS). Antimony, Se, Ag and Bi were not detected in any sample whereas the limits of detection (LODs) of these elements were 0.19, 0.40, 0.003 and 0.001 µg g(-1), respectively. Analysis of variance (ANOVA) revealed that the concentrations of Cd, Ti, Ni, As, Mo, U, Li and Be in yerba mate were not statistically different with regard to the country of origin, while those of the other investigated elements differed.

Imazethapyr and imazapic, bispyribac-sodium and penoxsulam: zooplankton and dissipation in subtropical rice paddy water.

Herbicides are very effective at eliminating weed and are largely used in rice paddy around the world, playing a fundamental role in maximizing yield. Therefore, considering the flooded environment of rice paddies, it is necessary to understand the side effects on non-target species. Field experiment studies were carried out during two rice growing seasons in order to address how the commonly-used herbicides imazethapyr and imazapic, bispyribac-sodium and penoxsulam, used at recommended dosage, affect water quality and the non-target zooplankton community using outdoor rice field microcosm set-up. The shortest (4.9 days) and longest (12.2 days) herbicide half-life mean, estimated of the dissipation rate (k) is shown for imazethapyr and bispyribac-sodium, respectively. Some water quality parameters (pH, conductivity, hardness, BOD5, boron, potassium, magnesium, phosphorus and chlorides) achieved slightly higher values at the herbicide treatment. Zooplankton community usually quickly recovered from the tested herbicide impact. Generally, herbicides led to an increase of cladocera, copepods and nauplius population, while rotifer population decreased, with recovery at the end of the experiment (88 days after herbicide treatment).

Toxic and micronutrient elements in organic, brown and polished rice in Brazil.

Concentration levels of As, Cd, Hg, Pb, Tl, Sn, Sb Co, Cu, Mn, Se, Zn, Cr, Ni and Mo in different types of rice cultivated in irrigated fields in Brazil were evaluated. Arsenic, Cd, Pb, Zn, Mn and Cu were found in higher concentrations in brown rice samples, suggesting the prevalence of these elements in the bran. Meanwhile, lower concentrations of Pb, Mo, Cr, Se and Co were found in parboiled rice. Organic rice did not differ of cultivated conventionally rice. Thallium, Hg and Sb were not detected in any rice sample whose limits of detection were 0.7 µg kg(-1), 2.5 µg kg(-1) and 8 µg kg(-1), respectively. The concentrations of the investigated elements were compared with those reported for polished rice and brown rice from other countries, unveiling concentrations in general at the same level for rice produced at non-contaminated sites.

Desorption electrospray ionization mass spectrometry (DESI-MS) applied to the speciation of arsenic compounds from fern leaves.

The different chemical forms of arsenic compounds, including inorganic and organic species, present distinct environmental impacts and toxicities. Desorption electrospray ionization mass spectrometry (DESI-MS) is an excellent technique for in situ analysis, as it operates under atmospheric pressure and room temperature and is conducted with no/minimal sample pretreatment. Aimed at expanding its scope, DESI-MS is applied herein for the quick and reliable detection of inorganic (arsenate--As(V): AsO4(3-) and arsenite--As(III): AsO2(-)) and organic (dimethylarsinic acid--DMA: (CH3)2AsO(OH) and disodium methyl arsonate hexahydrate: CH3AsO3·2Na·6H2O) arsenic compounds in fern leaves. Operational conditions of DESI-MS were optimized with DMA standard deposited on paper surfaces to improve ionization efficiency and detection limits. Mass spectra data for all arsenic species were acquired in both the positive and negative ion modes. The positive ion mode was shown to be useful in detecting both the organic and inorganic arsenic compounds. The negative ion mode was shown only to be useful in detecting As(V) species. Moreover, MS/MS spectra were recorded to confirm the identity of each arsenic compound by the characteristic fragmentation profiles. Optimized conditions of DESI-MS were applied to the analysis of fern leaves. LC-ICP-MS was employed to confirm the results obtained by DESI-MS and to quantify the arsenic species in fern leaves. The results confirmed the applicability of DESI-MS in detecting arsenic compounds in complex matrices.

Chemical phosphorus removal: a clean strategy for piggery wastewater management in Brazil.

The intensive production of animal protein is known to be an environmental polluting activity, especially if the wastewater produced is not managed properly. Swine production in Brazil is growing, and technologies to manage all pollutants present in the wastewater effluent are needed. This work presents a case of study of phosphorus (P) removal from piggery wastewater using Ca(OH)2, and demonstrates the feasibility of this strategy for P management. The effluent of a swine manure treatment plant was treated with Ca(OH)2. According to the addition of Ca(OH)2 the pH of the effluent ranged from 8.0 to 10.0. Different conditions of sludge dewatering were evaluated, and the chemical composition of sludge was investigated. Ion chromatography analysis of effluent samples showed that 92% of total P (TP) was present as soluble P (SP) whereas 75% of SP species were present as phosphate. The efficiency of P removal was typically 90% at pH 8.5 and higher than 98% at pH 10.5. The sludge was separated by sedimentation, centrifugation or filtration and dried. The TP content of dried sludge was 9.3% (w/w). X-ray diffraction analysis of the dry sludge showed the presence of amorphous compounds of Ca and P, which is an indication that the sludge obtained from the swine manure treatment has a potential for application as biofertilizer.

Effects of lead on the growth, lead accumulation and physiological responses of Pluchea sagittalis.

This work aimed to study the process of stress adaptation in root and leaves of different developmental stages (apex, middle and basal regions) of Pluchea sagittalis (Lam.) Cabrera plants grown under exposure to five Pb levels (0, 200, 400, 600 and 1000 µM) for 30 days. Pb concentration and content in roots, stems, and leaves of different developmental stages increased with external Pb level. Consumption of nutrient solution, transpiration ratio, leaf fresh weight, leaf area, and shoot length decreased upon addition of Pb treatments. However, dry weight of shoot parts and roots did not decrease upon addition of Pb treatments. Based on index of tolerance, the roots were much more tolerant to Pb than shoots. δ-aminolevulinic acid dehydratase activity was decreased by Pb treatments, whereas carotenoid and chlorophyll concentrations were not affected. Lipid peroxidation and hydrogen peroxide concentration both in roots and leaves increased with increasing Pb levels. Pb treatments increased ascorbate peroxidase activity in all plant parts, while superoxide dismutase activity increased in leaves and did not change in roots. Catalase activity in leaves from the apex shoot was not affected by Pb, but in other plant parts it was increased. Pb toxicity caused increase in non-protein thiol groups concentration in shoot parts, whereas no significant difference was observed in roots. Both root and shoot ascorbic acid concentration increased with increasing Pb level. Therefore, it seems that Pb stress triggered an efficient defense mechanism against oxidative stress in P. sagittalis but its magnitude was depending on the plant organ and of their physiological status. In addition, these results suggest that P. sagittalis is Pb-tolerant. In conclusion, P. sagittalis is able to accumulate on average 6730 and 550 µg Pb g(-1) dry weight, respectively, in the roots and shoot, a physiological trait which may be exploited for the phytoremediation of contaminated soils and waters.

Heavy crude oil sample preparation by pyrohydrolysis for further determination.

In this work, a method for chlorine determination in heavy crude oil using pyrohydrolysis for sample decomposition was developed. Chlorine was determined using ion chromatography (IC), inductively coupled plasma optical emission spectrometry (ICP OES) and inductively coupled plasma mass spectrometry. In this case an ICP-MS spectrometer equipped with a dynamic reaction cell (DRC-ICP-MS) was used. Heavy crude oil samples were homogenized and placed on a quartz holder, which was then introduced into the pyrohydrolysis apparatus. Nitric acid (0.2 mL) was used to assist the decomposition of the sample. Solutions with different concentrations of ammonium hydroxide were evaluated to collect the volatile chloride produced using pyrohydrolysis. A 0.75 mol L-1 ammonium hydroxide solution was chosen in view of the better accuracy achieved. Best precision and accuracy were obtained by heating the sample for 10 min up to 1000 °C. The limits of quantification (LOQs, 10σ) of chlorine measured by IC, ICP OES and DRC-ICP-MS were 4.5, 48 and 3.6 µg g-1, respectively. A sample amount of 300 mg and final volume of 20 mL were taken into account to calculate the LOQs. The relative standard deviation (RSD) was lower than 5% (4 replicates). Accuracy was evaluated by analysis of certified reference material (NIST 1634c) and comparison with the results obtained using the ASTM D6470 method. In both cases the results obtained using pyrohydrolysis were in agreement. The proposed method was suitable for heavy crude oil decomposition and chlorine determination by IC, ICP OES and DRC-ICP-MS. Three samples per hour can be processed using a simple and low-cost pyrohydrolysis apparatus. The pyrohydrolysis procedure is easy to perform, which is very attractive for routine analysis of crude oil, a very complex matrix. According to the authors' knowledge this is the first application of pyrohydrolysis for heavy crude oil sample preparation.

Cadmium and mineral nutrient accumulation in potato plantlets grown under cadmium stress in two different experimental culture conditions.

In order to evaluate the effect of cadmium (Cd(2+)) toxicity on mineral nutrient accumulation in potato (Solanum tuberosum L.), two cultivars named Asterix and Macaca were cultivated both in vitro and in hydroponic experiments under increasing levels of Cd(2+) (0, 100, 200, 300, 400 and 500 microM in vitro and 0, 50, 100, 150 and 200 microM in hydroponic culture). At 22 and 7 days of exposure to Cd(2+), for the in vitro and hydroponic experiment, respectively, the plantlets were separated into roots and shoot, which were analyzed for biomass as well as Cd(2+), and macro (Ca(2+), K(+) and Mg(2+)) and micronutrient (Cu(2+), Fe(2+), Mn(2+) and Zn(2+)) contents. In the hydroponic experiment, there was no reduction in shoot and root dry weight for any Cd(2+) level, regardless of the potato cultivar. In contrast, in the in vitro experiment, there was an increase in biomass at low Cd(2+) levels, while higher Cd(2+) levels caused a decrease. In general, Cd(2+) decreased the macronutrient and micronutrient contents in the in vitro cultured plantlets in both roots and shoot of cultivars. In contrast, the macronutrient and micronutrient contents in the hydroponically grown plantlets were generally not affected by Cd(2+). Our data suggest that the influence of Cd(2+) on nutrient content in potato was related to the level of Cd(2+) in the substrate, potato cultivar, plant organ, essential element, growth medium and exposure time.