Expression analysis of metallothioneins and mineral contents in tomato (Lycopersicon esculentum) under heavy metal stress.

BACKGROUND: Heavy metals are considered to be the most important pollutants in the contamination of soils; they adversely affect plant growth and development and cause some physiological and molecular changes. The contamination of agricultural soils by heavy metals has changed the mineral element content of vegetables. Plant metallothioneins (MTs) are thought to have the functional role in heavy metal homeostasis, and they are used as the biomarkers for evaluating environmental pollution. We aimed to evaluate the expression of MT isoforms (MT1, 2, 3 and 4) and some mineral element composition of tomato roots, leaves and fruits exposed to copper and lead. RESULTS: Heavy metal applications increased MT1 and MT2 gene expressions compared to the control in the tissues of tomato. The highest level of MT1 and MT2 transcripts was found in roots and leaves, respectively. The expression of MT3 is induced in roots, leaves and fruits except for Pb treatment in roots. MT4 expression increased in fruits; however, other tissues did not show a clear change. Our results indicated that Cu content was higher than Pb in all tissues of tomato. The lower doses of Cu (10 ppm) increased the content of Mg, Fe, Ca and Mn in roots. Pb generally increased the level of minerals in leaves and fruits, but it decreased Mg, Mn and Fe contents in roots. CONCLUSION: Both heavy metals not only moved to aerial parts but also caused alterations to mineral element levels. These results show that MT transcripts are regulated by Cu and Pb, and expression pattern changes to MT isoforms and tissue types. © 2016 Society of Chemical Industry.

Rapid extraction and reverse phase-liquid chromatographic separation of mercury(II) and methylmercury in fish samples with inductively coupled plasma mass spectrometric detection applying oxygen addition into plasma.

A simple and sensitive procedure was developed for extraction and speciation of mercury in fish. Species separation was accomplished with reverse phase-high performance liquid chromatography (HPLC) hyphenated to inductively coupled plasma mass spectrometry (ICP-MS). Oxygen addition into plasma allowed use of organic-rich mobile phase, achieving species separation in 4 min. Mercury species extraction was achieved by microwave exposure for 2 min at mild conditions (60°C, pH 2.0), avoiding necessity of neutralizing sample prior to injection in HPLC, and reducing number of sample preparation steps, analytical source of errors and inter conversion of species. Limit of detection for entire procedure was found to be 0.2 and 0.1 ng g(-1) for mercuric ion and methylmercury, respectively. The method was applied to certified reference materials (TORT-2 and DORM-2) and commercialized fish samples (Mullus barbatus, Sparus aurata, Trachurus mediterraneus, Mugil soiuy, Dicentrarchus labrax, and Pomatomus saltatrix) from Black Sea.

Influence of training frequency on serum concentrations of some essential trace elements and electrolytes in male swimmers.

Elemental fluctuations during physical performances have been a point of interest. This study was designed to investigate the effect of swimming frequency on serum concentrations of some trace elements (chromium, iron, copper, zinc, selenium) and electrolytes (sodium, magnesium, potassium, calcium). Three groups of different-level male swimmers were included in the study, as elite swimmers (n = 14), amateur swimmers (n = 11), and sedentary individuals (n = 10). Elite and amateur swimmer groups followed a 3-week training program. At the end of the period, all volunteers were subjected to a controlled swimming test, and blood samples were collected at the beginning of (pre-test), immediately after (post-test), and 1 h after this activity. Element concentrations were determined by inductively coupled plasma mass spectrometry using a dilute and shoot procedure. Apart from the swimming test applied, pre-test calcium and potassium levels were higher in elite swimmers compared to amateurs and controls. The difference in pre-test levels of these elements can be associated with adaptive mechanisms emerged by the frequent training. Regarding the test applied, changes in magnesium, calcium, copper, zinc, and selenium levels exhibited a common pattern in all study groups, with higher post-test serum concentrations. Another point of note was a drop of copper, zinc, and selenium levels at 1 h after the test in elite swimmers. The decrease in serum zinc was also observed in the other groups. Results highlight the value of regular control of elemental status to provide insight into transient effects and deficiencies.

Arsenic speciation in water and snow samples by adsorption onto PHEMA in a micro-pipette-tip and GFAAS detection applying large-volume injection.

A miniaturized solid phase extraction procedure has been developed for ultra-trace determination of inorganic arsenic species. Arsenic(III) as pyrrolidinedithiocarbamate complex was selectively adsorbed on 30 mg poly(hydroxyethyl methacrylate) (PHEMA) micro beads, which is simply packed into a micro-pipette-tip. The adsorbed arsenic was quantitatively eluted by 700 µL 0.25 M NH(3) and determined by graphite furnace atomic absorption spectrometry (GFAAS). Injection of larger volume (i.e., 50 µL v.s. conventional 10-20 µL) eluent into graphite furnace and the use of Mg(NO(3))(2) as chemical modifier have improved atomic absorption signal intensity (sensitivity as characteristic mass of 25 pg) and precision (RSD of 2.6%, c=10 µg L(-1), n=11). Total arsenic amount was determined after reduction of arsenic(V) to arsenic(III) by thiourea-HCl system. As(V) concentration was calculated by the difference between As(III) and total arsenic. The detection limit (3s) of the method was found as 10 ng L(-1) As(III) with an enrichment factor of 86. The relative standard deviation and relative error for six replicate determinations of 0.5 µg L(-1) As(III) were found to be 4.0% and -0.7%, respectively. The method was successfully applied to drinking water, snow and reference water (SEM-2011) samples. When the samples were spiked with 0.5 and 1.0 µg L(-1) As(III) and As(V), the recoveries varied between 96 and 100%.

Probing the metal-homeostatis effects of the administration of chromium(vi) to mice by ICP MS and size-exclusion chromatography-ICP MS.

Concentrations of chromium, copper, iron, manganese and zinc were determined in liver, kidney, brain, lung, heart and testis of mouse following intraperitoneal injection of hexavalent chromium [Cr(vi)] at a single dose of 8.0 mg Cr/kg. As result, chromium concentrations increased ca. 40-fold in liver and kidney and by a factor of 3-5 in all the other tissues. The homeostasis of Cu, Fe, Mn and Zn was also affected. The element molecular weight distribution was evaluated in the cytosols of the different mouse organs by size-exclusion chromatography (Superdex-75) with UV-VIS and ICP-MS detection. The administration of Cr(vi) resulted in differences in the elution profiles of Fe, Mn, Cu and Zn-protein complexes. Bioinduced Mn, Fe and Zn-binding proteins could be detected in some tissues, especially in liver and kidney. Different molecular weight fractions containing chromium were heartcut and submitted to tryptic digestion prior to MALDI MS analysis. Cr-peptide complexes could be obtained both in non-denaturing and in denaturing (in the presence of urea and DTT) conditions. They were isolated by size-exclusion chromatography with a smaller separation range (Superdex Peptide) but could not be identified by MALDI MS.

Matrix elimination method for the determination of precious metals in ores using electrothermal atomic absorption spectrometry.

Poly(N-(hydroxymethyl)methacrylamide)-1-allyl-2-thiourea) hydrogels, poly(NHMMA-ATU), were synthesized by gamma radiation using (60)Co gamma source in the ternary mixture of NHMMA-ATU-H(2)O. These hydrogels were used for the specific gold, silver, platinum and palladium recovery, pre-concentration and matrix elimination from the solutions containing trace amounts of precious metal ions. Elimination of inorganic matrices such as different transition and heavy metal ions, and anions was performed by adjusting the solution pH to 0.5 that was the selective adsorption pH of the precious metal ions. Desorption of the precious metal ions was performed by using 0.8 M thiourea in 3M HCl as the most efficient desorbing agent with recovery values more than 95%. In the desorption medium, thiourea effect on the atomic signal was eliminated by selecting proper pyrolysis and atomization temperatures for all precious metal ions. Precision and the accuracy of the results were improved in the graphite furnace-atomic absorption spectrometer (GFAAS) measurements by applying the developed matrix elimination method performing the adsorption at pH 0.5. Pre-concentration factors of the studied precious metal ions were found to be at least 1000-fold. Detection limits of the precious metal ions were found to be less than 10 ng L(-1) of the all studied precious metal ions by using the proposed pre-concentration method. Determination of trace levels of the precious metals in the sea-water, anode slime, geological samples and photographic fixer solutions were performed using GFAAS clearly after applying the adsorption-desorption cycle onto the poly(NHMMA-UTU) hydrogels.