The center of pathological (toxic) action of metals in an people organisms and a role of galvanic currents in its induction

Chronic influence of metals on an organism differs from their sharp toxic action. In some exposed to long chronic metals influence people appear chronic inflammatory diseases, including oncological, on a background of neurologic symptoms develop. Today the overwhelming majority of the population is more 40-50 years old have metal alloys in the form of dental artificial limbs and various implantates in a body. Because of metal corrosion in organisms of these people concentration of metal ions is created. Galvanic currents induced at presence of any metals in an organism, promote corrosion, and carry of metal ions inside organism. If these currents are strong (the potential difference more 150 MB), local inflammatory pathologies development in organism due to concentration in the center of metal ions.     

Metallic ion content and damage to the DNA in oral mucosa cells of children with fixed orthodontic appliances

Although the metal devices used in orthodontic treatments are manufactured highly resistance to corrosion, they may still suffer some localized corrosion resulting from the oral cavity conditions. The corrosion causes the release of metals from the alloys used for their manufacture. In this report, we evaluated the in vivo metal ions release of three alloys (stainless steel, titanium and nickel-free) usually used in the orthodontics treatments and its genotoxicity. We applied to 15 patients, between 12 and 16 years, 4 tubes and 20 brackets. Samples from oral mucosa were taken before the treatment and 30 days later. The concentration of the titanium, chromium, manganese, cobalt, nickel, molybdenum and iron were detected using inductively coupled plasma mass spectrometry (ICP-MS). The genotoxicity was measured with a comet assay (Olive moment). The oral mucosa cells in contact with the stainless steel alloy displayed the greatest titanium and manganese concentrations and those in contact with the nickel-free alloy presented the greatest concentration of chromium and iron. Both alloys, stainless steel and nickel-free, induced a higher DNA damage in the oral mucosa cells than the titanium alloy, in which the Olive moment was similar to controls. Based on the results of our study, we can conclude that titanium brackets and tubes are the most biocompatible of the three alloys.     

Effect of Heavy Metals （Cd, Cu） on the Gametophytes of Laminaria japonica Aresch

Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal （Cd） distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu up to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning （the first day） of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd-and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution of the ion in gametophytes and the results obtained were consistent with distribution determined using other methods. All results of the present study showed that gametophytes of L. japonica play a remarkable role as heavy metal decontaminators, especially with regard to Cd.     

THE INFLUENCE OF SIZE ON THE ACCUMULATED AMOUNTS OF METALS (Cu, Pb, Cd, Zn AND Ca) IN SIX SPECIES OF SLUG SAMPLED FROM A CONTAMINATED WOODLAND SITE

Heavy metal concentrations in 6 terrestrial slug species (Derocerasreticulatum, D. caruanae, Arionater, A. hortensis, A. subfuscusand Milax budapestensis) inhabiting a disused Pb/Zn mine sitewere measured by atomic absorption spectrophotometry. Relationshipsbetween dry weight: metal content and dry weight: metal concentration,respectively, were investigated by regression analysis beforeand after log transformation. Interspecies, as well as inter-metal differences in metal accumulationwere more effectively demonstrated by comparing metal content:dry body weight relationships, as opposed to metal concentration:dry body weight relationships. Regression analysis revealed differences in metal accumulationbetween the non-essential trace metals Pb/Cd and the metabolicallyessential trace metals Zn/Cu. Species differences in metal accumulation were more pronouncedwhen their coefficients of functional regression were compared,the greatest differences being observed for species belongingto different genera. However, the presence of a high level ofintrinsic variability in metal levels reduces the effectivenessand significance of the metal content: dry body weight expressions,and also detracts from the immediate prospect of using terrestrialslugs as biomonitors of environmental metal pollution. (Received 10 March 1989; accepted 25 November 1989)     

Burning question: Are there sustainable strategies to prevent microbial metal corrosion?

The global economic burden of microbial corrosion of metals is enormous. Microbial corrosion of iron-containing metals is most extensive under anaerobic conditions. Microbes form biofilms on metal surfaces and can directly extract electrons derived from the oxidation of Fe⁰ to Fe²⁺ to support anaerobic respiration. H₂ generated from abiotic Fe⁰ oxidation also serves as an electron donor for anaerobic respiratory microbes. Microbial metabolites accelerate this abiotic Fe⁰ oxidation. Traditional strategies for curbing microbial metal corrosion include cathodic protection, scrapping, a diversity of biocides, alloys that form protective layers or release toxic metal ions, and polymer coatings. However, these approaches are typically expensive and/or of limited applicability and not environmentally friendly. Biotechnology may provide more effective and sustainable solutions. Biocides produced with microbes can be less toxic to eukaryotes, expanding the environments for potential application. Microbially produced surfactants can diminish biofilm formation by corrosive microbes, as can quorum-sensing inhibitors. Amendments of phages or predatory bacteria have been successful in attacking corrosive microbes in laboratory studies. Poorly corrosive microbes can form biofilms and/or deposit extracellular polysaccharides and minerals that protect the metal surface from corrosive microbes and their metabolites. Nitrate amendments permit nitrate reducers to outcompete highly corrosive sulphate-reducing microbes, reducing corrosion. Investigation of all these more sustainable corrosion mitigation strategies is in its infancy. More study, especially under environmentally relevant conditions, including diverse microbial communities, is warranted.     

Pseudomonas fluorescens' view of the periodic table

Growth in a biofilm modulates microbial metal susceptibility, sometimes increasing the ability of microorganisms to withstand toxic metal species by several orders of magnitude. In this study, a high-throughput metal toxicity screen was initiated with the aim of correlating biological toxicity data in planktonic and biofilm cells to the physiochemical properties of metal ions. To this end, Pseudomonas fluorescens ATCC 13525 was grown in the Calgary Biofilm Device (CBD) and biofilms and planktonic cells of this microorganism were exposed to gradient arrays of different metal ions. These arrays included 44 different metals with representative compounds that spanned every group of the periodic table (except for the halogens and noble gases). The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum biofilm eradication concentration (MBEC) values were obtained after exposing the biofilms to metal ions for 4 h. Using these values, metal ion toxicity was correlated to the following ion-specific physicochemical parameters: standard reduction-oxidation potential, electronegativity, the solubility product of the corresponding metal–sulfide complex, the Pearson softness index, electron density and the covalent index. When the ions were grouped according to outer shell electron structure, we found that heavy metal ions gave the strongest correlations to these parameters and were more toxic on average than the other classes of the ions. Correlations were different for biofilms than for planktonic cells, indicating that chemical mechanisms of metal ion toxicity differ between the two modes of growth. We suggest that biofilms can specifically counter the toxic effects of certain physicochemical parameters, which may contribute to the increased ability of biofilms to withstand metal toxicity.     

Metal metabolism in the red alga Cyanidium caldarium and its relationship to metal tolerance

The unicellular red alga Cyanidium caldarium is tolerant to high levels of various metal ions. Cells of this alga cultured with divalent metal ions at 5 mM contained an elevated concentration of each metal, with the highest level for Zn followed by Mn > Ni > Cu. This order is in fair agreement with the toxicity levels reported previously, with the exception of Mn, which shows a toxicity level comparable to that of Ni. Transmission electron microscopy indicated the presence of electron-dense bodies in the algal cells, and elemental analysis by energy dispersive X-ray spectrometry showed high levels of Fe and P in these bodies. Accumulation of Zn was found in these particles in Zn-treated algal cells, whereas no such deposition was found for Cu, Ni, or Mn in cells treated with the respective metals. Although trapping of Zn in the intracellular bodies may contribute to reduction of metal activity in the cells, this effect can be overcome by high intracellular levels of Zn that result in a high degree of toxicity. The correlation between intracellular concentration and toxic levels of metal ions implies that the reduced incorporation of the metals is a major detoxification mechanism in this alga.     

Zinc fluxes and zinc transporter genes in chronic diseases

The group IIb metal zinc (Zn) is an essential dietary component that can be found in protein rich foods such as meat, seafood and legumes. Thousands of genes encoding Zn binding proteins were identified, especially after the completion of genome projects, an indication that a great number of biological processes are Zn dependent. Imbalance in Zn homeostasis was found to be associated with several chronic diseases such as asthma, diabetes and Alzheimer's disease. As it is now evident for most nutrients, body Zn status results from the interaction between diet and genotype. Zn ions cross biological membranes with the aid of specialized membrane proteins, belonging to the ZRT/IRT-related Proteins (ZIP) and zinc transporters (ZnT) families. The ZIPs are encoded by the Slc39A gene family and are responsible for uptake of the metal, ZnTs are encoded by the Slc30A genes and are involved in intracellular traffic and/or excretion. Both ZnTs and Zips exhibit unique tissue-specific expression, differential responsiveness to dietary Zn deficiency and excess, as well as to physiological stimuli via hormones and cytokines. Intracellular Zn concentration is buffered by metallothioneins (MTs), a class of cytosolic protein with high affinity for metals. Scattered information is available on the role of proteins responsible for regulating Zn fluxes in the onset and progression of chronic diseases. This paper reviews reports that link Zn transporter genes, their allelic variants and/or expression profiles in the context of specific diseases. Further investigation in this direction is very important, since Zn imbalance can result not only from insufficient dietary intake, but also from impaired activity of proteins that regulate Zn metabolism, thus contributing to multifactorial diseases.     

Effect of Heavy Metals (Cd, Cu) on the Gametophytes of Laminaria japonica Aresch

Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal (Cd) distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu up to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning (the first day) of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd- and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution of the ion in gametophytes and the results obtained were consistent with distribution determined using other methods.All results of the present study showed that gametophytes of L. japonica play a remarkable role as heavy metal decontaminators, especially with regard to Cd.     

黄河口盐地碱蓬湿地土壤-植物系统重金属污染评价

以黄河口盐地碱蓬湿地为例,评价了淹水和非淹水区湿地表层土壤As、Cd、Cu、Cr、Pb和Zn 6种重金属的污染程度及其在土壤-植物系统中的迁移、富集特征,分析了不同积水深度和土壤理化性质对研究区土壤重金属含量的影响.研究结果表明,与土壤或沉积物质量标准相比,黄河口盐地碱蓬湿地土壤受As和Cd污染最严重,而其它重金属污染较轻;非淹水土壤Cd、Cr和Zn含量高于淹水湿地,而As、Cu和Pb则较低;而且淹水土壤As含量随积水深度增加而呈下降趋势,但积水深度对其他重金属含量的影响不明显.相关性分析结果表明,按照受土壤关键影响因子的不同重金属(除As外)可以分为两类:第一类为Cd、Cr和Zn,这些重金属含量受土壤pH值和盐分影响较大,且相互间存在显著正相关关系,表明它们可能有相同的来源;第二类为Pb和Cu,它们受土壤pH值、盐分和有机质的影响,且Pb和Cu之间存在显著正相关关系.除Cr、Cu和Zn外,重金属在盐地碱蓬的根系内一般不发生显著富集,但绝大多数重金属都表现出地上部分的含量比根系更高的现象.     

Metal imaging in neurodegenerative diseases

Metal ions are known to play an important role in many neurodegenerative diseases including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and prion diseases. In these diseases, aberrant metal binding or improper regulation of redox active metal ions can induce oxidative stress by producing cytotoxic reactive oxygen species (ROS). Altered metal homeostasis is also frequently seen in the diseased state. As a result, the imaging of metals in intact biological cells and tissues has been very important for understanding the role of metals in neurodegenerative diseases. A wide range of imaging techniques have been utilized, including X-ray fluorescence microscopy (XFM), particle induced X-ray emission (PIXE), energy dispersive X-ray spectroscopy (EDS), laser ablation inductively coupled mass spectrometry (LA-ICP-MS), and secondary ion mass spectrometry (SIMS), all of which allow for the imaging of metals in biological specimens with high spatial resolution and detection sensitivity. These techniques represent unique tools for advancing the understanding of the disease mechanisms and for identifying possible targets for developing treatments. In this review, we will highlight the advances in neurodegenerative disease research facilitated by metal imaging techniques.     

Synthetic fluorescent sensors for studying the cell biology of metals

Metals are essential for sustaining all forms of life, but alterations in their cellular homeostasis are connected to severe human disorders, including cancer, diabetes and neurodegenerative diseases. Fluorescent small molecules that respond to metal ions in the cell with appropriate selectivity and sensitivity offer the ability to probe physiological and pathological consequences of the cell biology of metals with spatial and temporal fidelity. Molecular imaging of normal and abnormal cellular metal ion pools using these new chemical tools provides a host of emerging opportunities for visualizing, in real time, aspects of metal accumulation, trafficking, and function or toxicity in living systems. This review presents a brief survey of available synthetic small-molecule sensor types for fluorescence detection of cellular metals.     

Metalloregulation of Gram-positive pathogen physiology

Owing to the unique redox potential of transition metals, many of these elements serve important roles as cofactors in numerous enzymes. However, the reactive nature of metal becomes an intracellular threat when these ions are present in excess. Therefore, all organisms require mechanisms for sensing small fluctuations in metal levels to maintain a controlled balance of uptake, efflux, and sequestration. The ability to sense metal ion concentration is especially important for the survival of pathogenic bacteria because host organisms can both restrict access to essential metals from invading pathogens and utilize the innate toxicity of certain metals for bacterial killing. Host-induced metal ion fluctuations must be rapidly sensed by pathogenic bacteria so that they can activate metal transport systems, alter their physiology to accommodate differences in metal concentrations, and regulate the expression of virulence factors.     

Effect of Metal Cations on the Viscosity of a Pectin-Like Capsular Polysaccharide from the Cyanobacterium Microcystis flos-aquae C3-40

The properties of purified capsular polysaccharide from the cyanobacterium Microcystis flos-aquae C3-40 were examined by capillary viscometry. Capsule suspensions exhibited similar viscosities between pH 6 and 10 but were more viscous at pH <=4 than at pH 6 to 11. At pH 7, a biphasic effect of metal ion concentration on capsule viscosity was observed: (i) capsule viscosity increased with increasing metal ion concentration until a maximal viscosity occurred at a specific concentration that was a reproducible characteristic of each metal ion, and (ii) the viscosity decreased with further addition of that ion. Because the latter part of the biphasic curve was complicated by additional factors (especially the precipitation or gelation of capsule by divalent metal ions), the effects of various metal chlorides were compared for the former phase in which capsule viscosity increased in the presence of metal ions. Equivalent increases in capsule viscosity were observed with micromolar concentrations of divalent metal ions but only with 10 to 20 times greater concentrations of Na(sup+). The relative abilities of various metal salts to increase capsule viscosity were as follows: CdCl(inf2), Pb(NO(inf3))(inf2), FeCl(inf2) > MnCl(inf2) > CuCl(inf2), CaCl(inf2) >> NaCl. This pattern of metal efficacy resembles known cation influences on the structural integrity of capsule in naturally occurring and cultured M. flos-aquae colonies. The data are the first direct demonstration of an interaction between metal ions and purified M. flos-aquae capsule, which has previously been proposed to play a role in the environmental cycling of certain multivalent metals, especially manganese. The M. flos-aquae capsule and the plant polysaccharide pectin have similar sugar compositions but differ in their relative responses to various metals, suggesting that capsular polysaccharide could be a preferable alternative to pectin for certain biotechnological applications.     

In vitro assessment of the toxicity of metal compounds

This review has focused on several parameters related to the delivery of carcinogenic metal compounds to the cell nucleus as a basis for understanding the intermediates formed between metals and cellular components and the effect of these intermediates on DNA structure and function. Emphasis has been placed on metal interactions at the cellular membrane, including lipid peroxidation, metal interactions with glutathione and their relation to membrane injury, and metal effects on the membrane bound enzyme, Na⁺/K⁺ ATPase. Metal binding to metallothionein is also considered, particularly as related to transport and utilization of metal ions and to genetic defects in these processes exemplified in Menkes disease. The ability of cadmium to induce the synthesis of metallothionein more strongly than zinc is also discussed in relation to other toxic and carcinogenic metals. The effects of metal ions on purified DNA and RNA polymerase systems are presented with some of the recent studies using biological ligand-metal complexes. This review points out the importance of considering how metals affect in vitro systems when presented as ionic forms or complexed to relevant biological ligands.     

Research Advances on the Mechanisms of Heavy Metal Tolerance in Plants

Heavy metals impact on the cytoplasmic function in a number of different ways, principally by their binding to protein sulflhdryl groups, by producing a deficiency of essential ions and, eventually, by substituting the essemial ions. Other modes of toxicity are possible, including disruption of cell transport processes and oxidative damage by free radicals generated by metal redox cycling. Plants have developed a variety of biochemical defense strategies to prevent heavy metal poisoning. The possible defense mechanism in plant may involve: metal binding to cell walls, avoidance of uptake these toxic metal ions, reduction of heavy metal transport across the cell membrane, active efflux, compartmentalization and metal chelation. Phytochelatins that can tightly bind and sequester metals may play an important role in the accumulation of heavy metals and preventing them from entering the cell metabolic pathway, the rates of high molecular weight (HMW) metal phytochelatin complexes (Cd-Sa-complex) formation may be an important determinant of the plant tolerance. In addition, plants possess several antioxidant defense systems to protect themselves from the oxidative stress by heavy metals.     

Metals in cigarette smoke

Metals are vital for a huge number of physiological processes in the human body, but can also destroy health when the concentration is not within the physiologically favourable range. Cigarette smoking interferes with the carefully controlled metal homeostasis of the human body. This review focuses on the consequences of metal delivery to the human body by cigarette smoking and discusses the body's responses. The metal content of tobacco plants, smoke, the circulation, and various organs is discussed. Finally, we link individual cigarette smoke contained metals to the genesis of human diseases.     

The role of the root cell wall in the heavy metal tolerance ofAthyrium yokoscense

Cells of the roots ofA. yokoscense growing on metalliferous habitats were fractionated into their cell wall and cytoplasmic components. About 70–90% of the total copper, zinc and cadmium was located in the cell wall. Copper had a markedly greater affinity for the cell wall than zinc and cadmium, and was prevented from entering the cytoplasm. A large proportion of these heavy metals in the cell wall were exchanged as ions. The capacity of the cell wall for exchanging metal ions inA. yokoscense was higher than in other plants growing on metalliferous habitats. However, compared with different ferns unable to grow on metalliferous habitats, this capacity was not unique toA. yokoscense. Consequetly, the root cell wall ofA. yokoscense is considered to be an important site of metal ion storage and may play the role of an excretory organ for heavy metals. On the other hand, as proportion of the heavy metls was transported to the cytoplasm, where the metal content was much higher than the average for normal ferns. This would suggest thatA. yokoscense has another metabolic mechanism related to metal tolerance.     

Metallic dental material biocompatibility in osteoblastlike cells

Ions released from metallic dental materials used in orthodontic appliances could induce undesirable effects on cells and tissues. This study evaluates the biocompatibility of two of the most labile components of metallic dental alloys on osteoblastlike cells. The influence of protein and ions on metal dissolution properties is also investigated using different electrolyte solutions. Morphological alterations, cell growth, and differentiation of osteoblasts were assessed after exposure to pure metals (Ag, Cu, Pd, Au) and Ni−Ti alloy and correlated with the kinetics of elements released into the culture media. Results showed that Cu and Ag were the most cytotoxic elements and the other metals were biocompatible with the osteoblasts. The parameters of biocompatibility were correlated with the levels of ions detected into the culture media. Metal ions induced cell death through early mitosis arrest, apoptotic phenomena, and necrotic processes. Voltammograms showed that anions and proteins interfered in the corrosion process. Fetal bovine serum (FBS) strongly affected the electrochemical process, decreasing the oxidation rate of the metals. In conclusion, copper and silver ions showed a time-dependent low biocompatibility, which correlated with the concentration of released ions. The dissolution of the metallic materials was dependent on the composition of the simulated biological media.     

TRPM7 provides an ion channel mechanism for cellular entry of trace metal ions

Trace metal ions such as Zn(2+), Fe(2+), Cu(2+), Mn(2+), and Co(2+) are required cofactors for many essential cellular enzymes, yet little is known about the mechanisms through which they enter into cells. We have shown previously that the widely expressed ion channel TRPM7 (LTRPC7, ChaK1, TRP-PLIK) functions as a Ca(2+)- and Mg(2+)-permeable cation channel, whose activity is regulated by intracellular Mg(2+) and Mg(2+).ATP and have designated native TRPM7-mediated currents as magnesium-nucleotide-regulated metal ion currents (MagNuM). Here we report that heterologously overexpressed TRPM7 in HEK-293 cells conducts a range of essential and toxic divalent metal ions with strong preference for Zn(2+) and Ni(2+), which both permeate TRPM7 up to four times better than Ca(2+). Similarly, native MagNuM currents are also able to support Zn(2+) entry. Furthermore, TRPM7 allows other essential metals such as Mn(2+) and Co(2+) to permeate, and permits significant entry of nonphysiologic or toxic metals such as Cd(2+), Ba(2+), and Sr(2+). Equimolar replacement studies substituting 10 mM Ca(2+) with the respective divalent ions reveal a unique permeation profile for TRPM7 with a permeability sequence of Zn(2+) approximately Ni(2+) > Ba(2+) > Co(2+) > Mg(2+) >/= Mn(2+) >/= Sr(2+) >/= Cd(2+) >/= Ca(2+), while trivalent ions such as La(3+) and Gd(3+) are not measurably permeable. With the exception of Mg(2+), which exerts strong negative feedback from the intracellular side of the pore, this sequence is faithfully maintained when isotonic solutions of these divalent cations are used. Fura-2 quenching experiments with Mn(2+), Co(2+), or Ni(2+) suggest that these can be transported by TRPM7 in the presence of physiological levels of Ca(2+) and Mg(2+), suggesting that TRPM7 represents a novel ion-channel mechanism for cellular metal ion entry into vertebrate cells.