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Phytochelatins, a class of heavy-metal-binding peptides from plants, are functionally analogous to metallothioneins.

Phytochelatins are a class of heavy-metal-binding peptides previously isolated from cell suspension cultures of several dicotyledonous and monocotyledonous plants. These peptides consist of repetitive gamma-glutamylcysteine units with a carboxyl-terminal glycine and range from 5 to 17 amino acids in length. In the present paper we show that all plants tested synthesized phytochelatins upon exposure to heavy metal ions. No evidence for the occurrence of metallothionein-like proteins was found. All data so far obtained indicate that phytochelatins are involved in detoxification and homeostasis of heavy metals in plants and thus serve functions analogous to those of metallothioneins in animals and some fungi. Phytochelatins are induced by a wide range of metal anions and cations. Phytochelatin synthesis in suspension cultures was inhibited by buthionine sulfoximine, a specific inhibitor of gamma-glutamylcysteine synthetase (EC 6.3.2.2). This finding and kinetic studies of phytochelatin induction point to a synthesis from glutathione or its precursor, gamma-glutamylcysteine, in a sequential manner, thereby generating the set of homologous peptides.

Journal Article↗

Bacterial Community Structure and Function along a Heavy Metal Gradient.

The response of the planktonic, sediment, and epilithic bacterial communities to increasing concentrations of heavy metals was determined in a polluted river. None of the communities demonstrated a pollution-related effect on bacterial numbers (viable and total), heterotrophic activity, resistance to Pb or Cu, or species diversity as determined by either the Shannon-Wiener diversity index or rarefaction. The lack of correlation between concentrations of heavy metals and resistance in the sediment bacterial community was investigated and found to be due at least in part to the high pH of the river water and the resultant reduction in heavy metal toxicity. The three different communities demonstrated characteristic profiles based on the relative abundances of bacterial strains grouped according to functional similarities.

Journal Article↗

Atmospheric quality and distribution of heavy metals in Argentina employing Tillandsia capillaris as a biomonitor.

The atmospheric quality and distribution of heavy metals were evaluated throughout a wide region of Argentina. In addition, the biomonitor performance of Tillandsia capillaris Ruiz & Pav. f. capillaris was studied in relation to the accumulation of heavy metals and to its physiologic response to air pollutants. A sampling area of 50,000 km2 was selected in the central region of the Argentine Republic. This area was subdivided into grids of 25 x 25 km. Pools of T. capillaris, where present, were collected at each intersection point. From each pool three sub-samples were analyzed independently. Furthermore, five replicates were collected at 20% of the points in order to analyze the variability within the site. The content of Co, Cu, Fe, Ni, Mn, Pb and Zn was determined by Atomic Absorption Spectrometry. Chemical-physiological parameters were also determined to detect symptoms of foliar damage. Chlorophylls, phaeophytins, hydroperoxy conjugated dienes, malondialdehyde and sulfur were quantified in T. capillaris. Some of these parameters were used to calculate a foliar damage index. Data sets were evaluated by one-way ANOVA, correlation analysis, principal component analysis and mapping. Geographical distribution patterns were obtained for the different metals reflecting the contribution of natural and anthropogenic emission sources. According to our results it can be inferred that Fe, Mn and Co probably originated in the soil. For Pb, the highest values were found in the mountainous area, which can be attributed to the presence of Pb in the granitic rocks. Ni showed mainly an anthropogenic origin, with higher values found in places next to industrial centers. For Zn the highest values were in areas of agricultural development. The same was observed for Cu, whose presence could be related to the employment of pesticides. The foliar damage index distribution map showed that the central and southeastern zones were the ones where the major damage in the bioindicator was found. The central zone coincides with the city of Córdoba whereas the southeastern area is strictly agricultural, so the high values found there could be related to the use of pesticides.

Agriculture↗

Mechanisms of apoptosis in human retinal pigment epithelium induced by TNF-alpha in conditions of heavy metal ion deficiency.

PURPOSE: To investigate the mechanism underlying apoptosis in retinal pigment epithelium (RPE) induced by TNF-alpha in conditions of heavy metal ion deficiency. METHODS: Apoptotic morphology was assessed with Hoechst 33342. FITC-VAD-fmk or antibody specific to cleaved caspase 3 was used to detect in situ activated caspases or cleaved caspase 3, respectively. JC-1 and carboxylated dichlorodihydrofluorescein diacetate were used as probes to measure mitochondrial membrane potential (Deltapsi(m)) and intracellular reactive oxygen species (rOx). RESULTS: The apoptotic response of RPE cells was markedly enhanced when TNF-alpha plus actinomycin D (act-D) was coapplied with N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), a heavy metal ion chelator. The apoptosis was caspase dependent, and a blockade with cyclosporin A (CsA), an inhibitor of the mitochondrial permeability transition (MPT), but not FK506, a calcineurin inhibitor, abolished caspase activation and subsequent apoptosis, suggesting that apoptosis requires the MPT, and that caspase activation is downstream to the MPT. MPT, observed in situ as Deltapsi(m) loss, was prevented when cells were pretreated with either CsA or the pan-caspase inhibitor z-VAD-fmk. This result suggests that apoptotic signaling is initiated by the MPT and further amplified by downstream caspases, probably through a feedback loop. An increase in rOx was observed in cells exposed to TNF-alpha+act-D+TPEN, and rOx production did not require MPT or caspase activation. In addition, application of antioxidants significantly inhibited rOx production, Deltapsi(m) loss, and apoptosis. These data suggest that the rOx may play a role as a proapoptotic signal. CONCLUSIONS: The data suggest that intracellular heavy metal ions play a role in determining the apoptosis induction threshold of the inflammatory response to TNF-alpha in RPE.

Apoptosis↗

Magnetic response of soils and vegetation to heavy metal pollution--a case study.

Fast and cost-effective detection of industrial pollution can significantly promote its ecological, economic, and social assessment. A magnetometric method, used for qualitative determination of anthropogenic contamination, meets these requirements but needs further development in more quantitative terms. It could be used successfully in numerous cases when the heavy metals coexist with strongly magnetic iron oxide particles in the source dust. We present an integrated magnetic and geochemical study that examines the utility of magnetometric techniques for rapid, qualitative detection of metallic pollutants in soils and vegetation. The new aspect of our approach, in comparison with previously published articles on this subject, is the combined investigation (magnetic and geochemical) of both soils and vegetation, thus using an additional medium for employing the magnetometry as a pollution proxy at a site. The study area is a small (approximately 3 km2) region in the suburbs of Sofia (Bulgaria), with the main pollution source being a metallurgical factory. Soil samples have been taken from the topmost 20 cm from private gardens, located at different distances from the factory. Vegetation samples were taken from ryegrass (both leaves and roots) and leaves from two kinds of deciduous trees (maple and acacia). The results show that both vegetation and soils are characterized by enhanced magnetic properties, compared to background material, which is due to the presence of magnetite particles of anthropogenic origin accompanying heavy metal emissions. SEM images and microprobe analyses reveal the presence of a significant amount of particles, containing heavy metals (including iron) in vegetation samples taken close to the main pollution source. Correlation analyses show a statistically significant link (correlation coefficients ranging from 0.6 to 0.7) between magnetic susceptibility and the main heavy metals (Cu, Zn, Pb) in soil samples, indicating that the magnetic susceptibility can provide a proxy method for identifying the relative contribution of industrial pollution in soils and vegetation, that is reliable, inexpensive, and less time-consuming than standard chemical analyses.

Environmental Monitoring↗

Use of diammonium phosphate to reduce heavy metal solubility and transport in smelter-contaminated soil.

Phosphate treatments can reduce metal dissolution and transport from contaminated soils. However, diammonium phosphate (DAP) has not been extensively tested as a chemical immobilization treatment. This study was conducted to evaluate DAP as a chemical immobilization treatment and to investigate potential solids controlling metal solubility in DAP-amended soils. Soil contaminated with Cd, Pb, Zn, and As was collected from a former smelter site. The DAP treatments of 460, 920, and 2300 mg P kg-1 and an untreated check were evaluated using solute transport experiments. Increasing DAP decreased total metal transported. Application of 2300 mg P kg-1 was the most effective for immobilizing Cd, Pb, and Zn eluted from the contaminated soil. Metal elution curves fitted with a transport model showed that DAP treatment increased retardation (R) 2-fold for Cd, 6-fold for Zn, and 3.5-fold for Pb. Distribution coefficients (Kd) increased with P application from 4.0 to 9.0 L kg-1 for Cd, from 2.9 to 10.8 L kg-1 for Pb, and from 2.5 to 17.1 L kg-1 for Zn. Increased Kd values with additional DAP treatment indicated reduced partitioning of sorbed and/or precipitated metal released to mobile metal phases and a concomitant decrease in the concentration of mobile heavy metal species. Activity-ratio diagrams indicated that DAP decreased solution Cd, Pb, and Zn by forming metal-phosphate precipitates with low solubility products. These results suggest that DAP may have potential for protecting water resources from heavy metal contamination near smelting and mining sites.

Industry↗

Biological and health implications of toxic heavy metal and essential trace element interactions.

Human civilization and a concomitant increase in industrial activity has gradually redistributed many toxic metals from the earth's crust to the environment and increased the possibility of human exposure. Among the various toxic elements, heavy metals cadmium, lead, and mercury are specially prevalent in nature due to their high industrial use. These metals serve no biological function and their presence in tissues reflects contact of the organism with its environment. They are cumulative poison, and are toxic even at low dose. Studies of metabolism and toxicity of these elements have revealed important interactions between them and some essential dietary elements like calcium, zinc, iron, selenium, copper, chromium, and manganese. In general, a deficiency of these essential elements increases toxicity of heavy metals, whereas an excess appears to be protective. While most of the observations are on laboratory animals, limited human data are in agreement with the results of animal experiments. These suggest that the dietary presence of the essential elements may contribute to the protection of man and animal from the effects of heavy metal exposure, while their deficiency may increase toxicity. Appropriate dietary manipulation thus may be valuable in the prevention and treatment of heavy metal toxicity.

Animals↗

Development of an inverse method to identify the kinetics of heavy metal release during waste incineration in fluidized bed.

This paper deals with the emission of heavy metals (HM) during the incineration of municipal solid waste in a fluidized bed reactor. This study focused on the development of a general method to identify the kinetics of vaporization of heavy metals from the on-line analysis of exhaust gas. This method is an inverse method, which requires only the time evolution of the HM concentration in exhaust gases (experimental data) and a global bubbling bed model developed for transient conditions at the reactor scale. First, a lab-scale fluidized bed incinerator was set-up to simulate the HM release during the thermal treatment of metal-spiked model wastes. A specific on-line analysis system based on ICP-OES was developed to measure in real time the variation of the relative concentration of HM in exhaust gases. Then, a two-phase flow bubbling bed model was developed and validated to calculate the kinetics of vaporization of HM from its measured concentration time profile in the outlet gas. The technique was first validated with model waste (metal-spiked mineral matrices), thus enabling at each time both solid sampling for measuring the HM vaporization kinetic and on-line analysis for measuring the HM concentration in the outlet gas. The inverse method was then applied to realistic artificial wastes (derived from real wastes) to identify the HM vaporization kinetics from the on-line analysis results.

Aluminum Oxide↗

Effects of high temperature melting on fractionation of heavy metals in sewage sludge.

Sewage sludge from aerobic treatment plant was found to contain high amounts of heavy metals. Research was carried out to investigate the speciation and leaching behavior of heavy metals when using high temperature melting technology for treatment. This was achieved by conducting a sequential chemical extraction procedure and EP-TOX leaching test. The thermal treatment led to increased shift of metals from organic fraction to residual fraction, indicating that the thermal treatment caused metals in sewage sludge to become stable. Furthermore, results from leaching test revealed that metals were not leached from the final product after thermal treatment and this was verified using US EPA standard limits. Results from this study indicated that melting technology could convert the sludge to product that can be either reused or landfilled without an adverse environmental impact.

Biological Availability↗

Occupational exposure to heavy metals: DNA damage induction and DNA repair inhibition prove co-exposures to cadmium, cobalt and lead as more dangerous than hitherto expected.

Co-exposure to cadmium, cobalt, lead and other heavy metals occurs in many occupational settings, such as pigment and batteries production, galvanization and recycling of electric tools. However, little is known about interactions between several heavy metals. In the present study we determined DNA single strand break (DNA-SSB) induction and repair capacity for 8-oxoguanine in mononuclear blood cells of 78 individuals co-exposed to cadmium (range of concentrations in air: 0.05-138.00 micro g/m(3)), cobalt (range: 0-10 micro g/m(3)) and lead (range: 0-125 micro g/m(3)). Exposure to heavy metals was determined in air, blood and urine. Non-parametric correlation analysis showed a correlation between cadmium concentrations in air with DNA-SSB (P = 0.001, R = 0.371). Surprisingly, cobalt air concentrations correlated even better (P < 0.001, R = 0.401), whereas lead did not correlate with DNA-SSB. Logistic regression analysis including 11 possible parameters of influence resulted in a model showing that cobalt in air, cadmium in air, cadmium in blood and lead in blood influence the level of DNA-SSB. The positive result with cobalt was surprising, since exposure levels were much lower compared with the TRK-value of 100 micro g/m(3). To examine, whether the positive result with cobalt is stable, we applied several logistic regression models with two blocks, where all factors except cobalt were considered preferentially. All strategies resulted in the model described above. Logistic regression analysis considering also all possible interactions between the relevant parameters of influence finally resulted in the following model: Odds ratio = 1.286(Co in air) x 1.040(Cd in air) x 3.111(Cd in blood) x 0.861(Pb in air) x 1.023(Co in air x Pb in air). This model correctly predicts an increased level of DNA-SSB in 91% of the subjects in our study. One conclusion from this model is the existence of more than multiplicative effects for co-exposures of cadmium, cobalt and lead. For instance increasing lead air concentrations from 1.6 to 50 micro g/m(3) in the presence of constant exposures to cobalt and cadmium (8 micro g/m(3) and 3.8 micro g/m(3)) leads to an almost 5-fold increase in the odds ratio, although lead alone does not increase DNA-SSB. The mechanism behind these interactions might be repair inhibition of oxidative DNA damage, since a decrease in repair capacity will increase susceptibility to reactive oxygen species generated by cadmium or cobalt. Indeed, repair of 8-oxoguanine decreased with increasing exposures and inversely correlated with the level of DNA-SSB (P = 0.001, R = -0.427). Protein expression patterns of individuals exposed to cobalt concentrations of approximately 10 micro g/m(3) were compared with those of unexposed individuals using two-dimensional gel electrophoresis. Qualitative and apparent quantitative alterations in protein expression were selective and certainly occurred in <0.1% of all proteins. In conclusion, the hazard due to cobalt exposure - that has been classified only as IIB by the IARC - seems to be underestimated, especially when individuals are co-exposed to cadmium or lead. Co-exposure may cause genotoxic effects, even if the concentrations of individual heavy metals do not exceed TRK-values.

Alcohol Drinking↗

Determination of the heavy metal binding capacity of aquatic samples using MetPLATE: a preliminary study.

MetPLATE, a microbial toxicity test which is specific for heavy metal toxicity, was used to rapidly determine the heavy metal binding capacity (HMBC) of a wide range of surface waters from Florida and Georgia. HMBC determines the impact of physicochemical factors on metal bioavailability and toxicity. The new developed protocol, using MetPLATE as the toxicity assay, showed that HMBC varied from 1.7 to 39.2 for Cd whereas the ranges for Cu and Ag were < 1-11.9 and < 1-2.1, respectively. The effect of seasons on HMBC was determined using samples from the Hogtown Creek, Gainesville, FL, and from the St John's river in Jacksonville, FL. Both surface waters displayed the highest HMBC during the Fall season. Preliminary examination of the limited data set confirms that HMBC may be influenced by the concentration of dissolved organic carbon (DOC). These preliminary data show that the impact of physical and chemical parameters on the toxicity of metals in aquatic environments can be rapidly assessed using rapid and low-cost microbiotests.

Biological Availability↗

Determination of traces of heavy metals (Mn, Cu, Zn, Cd and Pb) in microsamples of teeth material by ETV-ICP-MS.

Heavy metal levels in tooth compartments would appear to be a suitable indicator of long-term exposure. A method has been developed to assess the heavy metal levels (Mn, Cu, Zn, Cd and Pb) in tooth compartments like dentin and enamel. For the simultaneous determination of these elements in small dentin sample amounts (in some cases less than 1 mg) inductively coupled plasma mass spectrometry with electrothermal vaporization was the method of choice. Sample preparation, method development and first results of the application of the method for the analysis of tooth compartments have been described. Variation in the elemental concentrations depending on the kind of tooth and the sampling position have been observed. The method is useful for epidemiological studies of the heavy metal exposure of children.

Journal Article↗

Heavy metals in traditional Indian remedies.

The growing popularity of traditional Indian remedies necessitates a critical evaluation of risks associated with their use. This systematic review aims at summarising all available data relating to the heavy metal content in such remedies. Computerised literature searches were carried out to identify all articles with original data on this subject. Fifteen case reports and six case series were found. Their collective results suggest that heavy metals, particularly lead, have been a regular constituent of traditional Indian remedies. This has repeatedly caused serious harm to patients taking such remedies. The incidence of heavy metal contamination is not known, but one study shows that 64% of samples collected in India contained significant amounts of lead (64% mercury, 41% arsenic and 9% cadmium). These findings should alert us to the possibility of heavy metal content in traditional Indian remedies and motivate us to consider means of protecting consumers from such risks.

Adult↗

Ecophysiological responses of Empetrum nigrum to heavy metal pollution.

Chlorophyll, organic (citric and malic acids) and abscisic acid (ABA) contents and stem water potential were measured to indicate possible physiological effects of heavy metal deposition on Empetrum nigrum L. (crowberry). The leaves and stems of E. nigrum were collected at distances of 0.5 and 8 km from the Cu-Ni smelter at Harjavalta, south-west Finland. All the investigated parameters were clearly affected by heavy metal emissions. Chlorophyll contents in the leaves and organic acid contents in the leaves and stems were lower close to the emission source. Generally found increase in organic acid contents with increasing Ni concentrations was not found, which might be due to the lower production of organic acids measured by decreased photosynthesis near the smelter. In contrast, ABA contents in stems and leaves in general, were higher in plants growing 0.5 km from the pollution source. Close to the smelter the stem water potential of E. nigrum was less negative during the day but more negative during the night. These results suggest that smelter emissions have a negative effect on the ecophysiology of E. nigrum even though it is considered to be a tolerant species to heavy metals.

Chlorophyll↗