Heavy metal uptake capacities by the common freshwater green alga Cladophora fracta

Macroalgae have received much attention for heavy metal removal in treatment of domestic wastewater. In this report, the uptake capacity of a common freshwater green alga, Cladophora fracta, for heavy metal ions (copper, zinc, cadmium, and mercury) was evaluated. The equilibrium adsorption capacities were 2.388?mg Cu²⁺, 1.623?mg Zn²⁺, 0.240?mg Cd²⁺, and 0.228?mg Hg²⁺ per gram of living algae at 18°C and pH?5.0. The removal efficiency for Cu²⁺, Zn²⁺, Cd²⁺, and Hg²⁺ were 99, 85, 97, and 98%, respectively. Greater removal efficiency was achieved when the concentrations of metal ions were at very low level. The results indicated that living algae are suitable for removal and recovery of heavy metal ions from aqueous solutions and can be a potential tool to treat industrial wastewater.     

Development of a fluorescent transgenic zebrafish biosensor for sensing aquatic heavy metal pollution

We report a transgenic zebrafish (Danio rerio) designed to respond to heavy metals using a metal-responsive promoter linked to a fluorescent reporter gene (DsRed2). The metallothionein MT-Ia1 promoter containing metal-responsive elements was derived from the Asian green mussel, Perna viridis. The promoter is known to be induced by a broad spectrum of heavy metals. The promoter-reporter cassette cloned into the Tol2 transposon vector was microinjected into zebrafish embryos that were then reared to maturity. A transgene integration rate of 28 % was observed. The confirmed transgenics were mated with wild-type counterparts, and pools of F₁ embryos were exposed to sub-lethal doses of Cd²⁺, Cu²⁺, Hg²⁺, Pb²⁺ and Zn²⁺. The red fluorescence response of zebrafish embryos was observed 8 h post- exposure to these sub-lethal doses of heavy metals using a fluorescence microscope. Reporter expression estimated by real-time PCR revealed eightfold, sixfold and twofold increase on exposure to highest concentrations of Hg²⁺, Cd²⁺ and Cu²⁺, while Pb²⁺ and Zn²⁺ had no effect. This biosensor could be a first-level screening method for confirming aquatic heavy metal bio-toxicity to eukaryotes.     

A new fluorescent chemosensor for Hg2+ in aqueous solution

We prepared an aminothiourea‐derived Schiff base (DA) as a fluorescent chemosensor for Hg²⁺ ions. Addition of 1 equiv of Hg²⁺ ions to the aqueous solution of DA gave rise to an obvious fluorescence enhancement and the subsequent addition of more Hg²⁺ induced gradual fluorescence quenching. Other competing ions, including Pb²⁺, Cd²⁺, Cr³⁺, Zn²⁺, Fe²⁺, Co³⁺, Ni²⁺, Ca²⁺, Mg²⁺, K⁺ and Na⁺, did not induce any distinct fluorescence changes, indicating that DA can selectively detect Hg²⁺ ions in aqueous solution. Copyright © 2012 John Wiley & Sons, Ltd.     

A new rhodamine‐based fluorescent chemodosimeter for mercuric ions in water media

A new rhodamine–ethylenediamine–nitrothiourea conjugate (RT) was synthesized and its sensing property as a fluorescent chemodosimeter toward metal ions was explored in water media. Analytical results from absorption and fluorescence spectra revealed that the addition of Hg²⁺ ions to the aqueous solution of the chemodosimeter RT caused a distinct fluorescence OFF–ON response with a remarkable visual color change from colorless to pink; however, no clear spectral and color changes were observed from other metal ions including: Zn²⁺, Cu²⁺, Cd²⁺, Pb²⁺, Ag⁺, Fe²⁺, Cr³⁺, Co³⁺, Ni²⁺, Ca²⁺, Mg²⁺, K⁺ and Na⁺. The sensing results and the molecular structure suggested that a Hg²⁺‐induced a desulfurization reaction and cyclic guanylation of the thiourea moiety followed by ring‐opening of the rhodamine spirolactam in RT are responsible for a distinct fluorescence turn‐on signal, indicating that RT is a remarkably sensitive and selective chemodosimeter for Hg²⁺ ions in aqueous solution. Hg²⁺ within a concentration range from 0.1 to 25 μM can be determined using RT as a chemodosimeter and a detection limit of 0.04 μM is achieved. Copyright © 2014 John Wiley & Sons, Ltd.     

Queuine-containing isoacceptor of tyrosine tRNA in Drosophila melanogaster: Alteration of levels by divalent cations

Dietary cadmium causes the queuine-containing, Q(+), isoacceptors to increase relative to the guanine-containing, Q(?), ones of tRNA^Tyr, tRNA^His and tRNA^Asp of Drosophila melanogaster. Of the other divalent cations examined, Sr²⁺, Ni²⁺, Cu²⁺, Zn²⁺ and Hg²⁺, only Hg²⁺ failed to cause an increase in Q(+)tRNA^Tyr. For these results, all pre-adult stages of the organism were spent on media containing the divalent ions. Adult flies that had developed on a normal diet also responsed to divalent ions; Hg²⁺ as well as Cd²⁺, Sr²⁺ and Zn²⁺ caused an increase in Q(+)tRNA^Tyr in 4 days. Using adult flies, the rate of the response was measured; when placed on a Cd²⁺-containing diet, they formed significantly more Q(+)tRNA^Tyr within 24 h as compared to adults on a normal diet. Whether the queuine is derived from the diet or from de novo synthesis is yet to be determined. Since the metal ions represent a range of values in the ‘hard-soft’ classification, different sites of reaction are expected, yet for Drosophila a common result is an alteration in the ratio of Q(+) and Q(?) isoacceptors of these tRNAs. The transition to Q(+)tRNA may be an early indication of the metabolic imbalances resulting from the presence of the divalent cation.     

Multimode selective detection of mercury by chiroptical fluorescent sensors based on methionine/cysteine

Two multimode Hg(II) sensors, L‐MethBQA and L‐CysBQA, were obtained by fusing methionine or S‐methyl cysteine, into a bis‐quinolyl amine‐based chiral podand scaffold. Quinolyl groups serve as the fluorophore and possess nitrogen lone pairs capable of chelating metal ions. On exposure to Hg²⁺ or Zn²⁺, these sensors show signal enhancement in fluorescence. However, Cu²⁺ quenches their fluorescence in 30:70 acetontrile/water. L‐CysBQA complexes with Hg²⁺, producing an exciton‐coupled circular dichroism spectrum with the opposite sign to the one that is produced by Cu²⁺ or Zn²⁺ complexation. L‐CysBQA binds Hg²⁺ more strongly than Zn²⁺ and is shown to differentiate Hg²⁺ from other metal ions, such as Zn²⁺, Cu²⁺, Ni²⁺, and Pb²⁺, exceptionally well. The synergistic use of relatively soft sulfur, quinoline‐based chiral ligands and chiroptically enhanced fluorescence detection results in high sensitivity and selectivity for Hg²⁺. Chirality, 2011. © 2011 Wiley‐Liss, Inc.     

Calcium signaling mediates the response to cadmium toxicity in Saccharomyces cerevisiae cells

The involvement of Ca²⁺ in the response to high Mn²⁺, Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺, Cd²⁺, and Hg²⁺ was investigated in Saccharomyces cerevisiae. The yeast cells responded through a sharp increase in cytosolic Ca²⁺ when exposed to Cd²⁺, and to a lesser extent to Cu²⁺, but not to Mn²⁺, Co²⁺, Ni²⁺, Zn²⁺, or Hg²⁺. The response to high Cd²⁺ depended mainly on external Ca²⁺ (transported through the Cch1p/Mid1p channel) but also on vacuolar Ca²⁺ (released into the cytosol through the Yvc1p channel). The adaptation to high Cd²⁺ was influenced by perturbations in Ca²⁺ homeostasis. Thus, the tolerance to Cd²⁺ often correlated with sharp Cd²⁺-induced cytosolic Ca²⁺ pulses, while the Cd²⁺ sensitivity was accompanied by the incapacity to rapidly restore the low cytosolic Ca²⁺.     

A bacterial view of the periodic table: genes and proteins for toxic inorganic ions

Essentially all bacteria have genes for toxic metal ion resistances and these include those for Ag⁺, AsO ⁻₂ , AsO ³⁻₄ , Cd²⁺, Co²⁺, CrO ²⁻₄ , Cu²⁺, Hg²⁺, Ni²⁺, Pb²⁺, TeO ²⁻₃ , Tl⁺ and Zn²⁺. The largest group of resistance systems functions by energy-dependent efflux of toxic ions. Fewer involve enzymatic transformations (oxidation, reduction, methylation, and demethylation) or metal-binding proteins (for example, metallothionein SmtA, chaperone CopZ and periplasmic silver binding protein SilE). Some of the efflux resistance systems are ATPases and others are chemiosmotic ion/proton exchangers. For example, Cd²⁺-efflux pumps of bacteria are either inner membrane P-type ATPases or three polypeptide RND chemiosmotic complexes consisting of an inner membrane pump, a periplasmic-bridging protein and an outer membrane channel. In addition to the best studied three-polypeptide chemiosmotic system, Czc (Cd²⁺, Zn²⁺, and Co²), others are known that efflux Ag⁺, Cu⁺, Ni²⁺, and Zn²⁺. Resistance to inorganic mercury, Hg²⁺ (and to organomercurials, such as CH₃Hg⁺ and phenylmercury) involve a series of metal-binding and membrane transport proteins as well as the enzymes mercuric reductase and organomercurial lyase, which overall convert more toxic to less toxic forms. Arsenic resistance and metabolizing systems occur in three patterns, the widely-found ars operon that is present in most bacterial genomes and many plasmids, the more recently recognized arr genes for the periplasmic arsenate reductase that functions in anaerobic respiration as a terminal electron acceptor, and the aso genes for the periplasmic arsenite oxidase that functions as an initial electron donor in aerobic resistance to arsenite.

     

Effects of heavy metals on seed germination and early seedling growth of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Seed is a developmental stage that is highly protective against external stresses in the plant life cycle. In this study, we analyzed toxicity of essential (Cu²⁺ and Zn²⁺) and non-essential heavy metals (Hg²⁺, Pb²⁺ and Cd²⁺) on seed germination and seedling growth in the model species Arabidopsis. Our results show that seedling growth is more sensitive to heavy metals (Hg²⁺, Pb²⁺, Cu²⁺ and Zn²⁺) in comparison to seed germination, while Cd²⁺ is the exception that inhibited both of these processes at similar concentrations. To examine if toxicity of heavy metals is altered developmentally during germination, we incubated seeds with Hg²⁺ or Cd²⁺ only for a restricted period during germination. Hg²⁺ displayed relatively strong toxicity at period II (12–24 h after imbibition), while Cd²⁺ was more effective to inhibit germination at period I (0–12 h after imbibition) rather than at period II. The observed differences are likely to be due in part to selective uptake of different ions by the intact seed, because isolated embryos (without seed coat and endosperm) are more sensitive to both Hg²⁺ and Cd²⁺ at period I. We assessed interactive toxicity between heavy metals and non-toxic cations, and found that Ca²⁺ was able to partially restore the inhibition of seedling growth by Pb²⁺ and Zn²⁺.     

Substrate Profile and Metal-ion Selectivity of Human Divalent Metal-ion Transporter-1

Divalent metal-ion transporter-1 (DMT1) is a H⁺-coupled metal-ion transporter that plays essential roles in iron homeostasis. DMT1 exhibits reactivity (based on evoked currents) with a broad range of metal ions; however, direct measurement of transport is lacking for many of its potential substrates. We performed a comprehensive substrate-profile analysis for human DMT1 expressed in RNA-injected Xenopus oocytes by using radiotracer assays and the continuous measurement of transport by fluorescence with the metal-sensitive PhenGreen SK fluorophore. We provide validation for the use of PhenGreen SK fluorescence quenching as a reporter of cellular metal-ion uptake. We determined metal-ion selectivity under fixed conditions using the voltage clamp. Radiotracer and continuous measurement of transport by fluorescence assays revealed that DMT1 mediates the transport of several metal ions that were ranked in selectivity by using the ratio I_max/K_0.5 (determined from evoked currents at −70 mV): Cd²⁺ > Fe²⁺ > Co²⁺, Mn²⁺ ≫ Zn²⁺, Ni²⁺, VO²⁺. DMT1 expression did not stimulate the transport of Cr²⁺, Cr³⁺, Cu⁺, Cu²⁺, Fe³⁺, Ga³⁺, Hg²⁺, or VO⁺. ⁵⁵Fe²⁺ transport was competitively inhibited by Co²⁺ and Mn²⁺. Zn²⁺ only weakly inhibited ⁵⁵Fe²⁺ transport. Our data reveal that DMT1 selects Fe²⁺ over its other physiological substrates and provides a basis for predicting the contribution of DMT1 to intestinal, nasal, and pulmonary absorption of metal ions and their cellular uptake in other tissues. Whereas DMT1 is a likely route of entry for the toxic heavy metal cadmium, and may serve the metabolism of cobalt, manganese, and vanadium, we predict that DMT1 should contribute little if at all to the absorption or uptake of zinc. The conclusion in previous reports that copper is a substrate of DMT1 is not supported.     

A highly selective fluorescence and absorption sensor for rapid recognition and detection of Cu2+ ions in aqueous solution and film

A fluorescence and absorption chemosensor (SAAT) based on 5-(hydroxymethyl)-salicylaldehyde (SA) and o-aminothiophenol (AT) was designed and synthesized. SAAT in DMSO–HEPES (20.0 mM, v/v, 1:99, pH = 7.0) solution shows a highly selective and sensitive absorption and an ‘on–off’ fluorescence response to Cu²⁺ ions in aqueous solutions over all other competitive metal ions including Na⁺, Ag⁺, Ba²⁺, Ca²⁺, Cd²⁺, Mg²⁺, Zn²⁺, Cr³⁺, Al³⁺, Hg²⁺, K⁺, Mn²⁺, Ni²⁺, Sr²⁺, Tb³⁺ and Co²⁺. SAAT exhibits ratiometric absorption sensing ability for Cu²⁺ ions. Importantly, SAAT also can sense Cu²⁺ ions using fluorescence quenching, the fluorescence intensity of SAAT showed a good linear relationship with Cu²⁺ concentration, and the detection limit of Cu²⁺ was 0.34 μM. The results of Job's plot, Benesi–Hildebrand plot, mass spectra, and density functional theory calculations confirmed that the selective absorption and fluorescence response were attributed to the formation of a 1:1 complex between SAAT and Cu²⁺. SAAT in test film could identify Cu²⁺ in water samples using the intuitive fluorescence colour change under a UV lamp. SAAT has great application value as a selective and sensitive chemosensor to discriminate and detect Cu²⁺ ions.     

Bacterial resistances to toxic metal ions - a review

Bacterial plasmids encode resistance systems for toxic metal ions, including Ag⁺, AsO₂^-, AsO₄^3-, Cd²⁺, Co²⁺, CrO₄^2-, Cu²⁺ Hg²⁺, Ni²⁺, Pb²⁺, Sb³⁺, TeO₃^2-, Tl⁺ and Zn²⁺. The function of most resistance systems is based on the energy-dependent efflux of toxic ions. Some of the efflux systems are ATPases and others are chemiosmotic cation/proton antiporters. The Cd²⁺-resistance ATPase of Gram-positive bacteria (CadA) is membrane cation pump homologous with other bacterial, animal and plant P-type ATPases. CadA has been labeled with ³²P from [α-³²p]ATP and drives ATP-dependent Cd²⁺ (and Zn²⁺) uptake by inside-out membrane vesicles (equivalent to efflux from whole cells). Recently, isolated genes defective in the human hereditary diseases of copper metabolism, namely Menkes syndrome and Wilson's disease, encode P-type ATPases that are more similar to bacterial CadA than to other ATPases from eukaryotes. The arsenic resistance efflux system transports arsenite [As(III)], alternatively using either a double-polypeptide (ArsA and ArsB) ATPase or a single-polypeptide (ArsB) functioning as a chemiosmotic transporter. The third gene in the arsenic resistance system, arsC, encodes an enzyme that converts intracellular arsenate [As(V)] to arsenite [As(III)], the substrate of the efflux system. The triple-polypeptide Czc (Cd²⁺, Zn²⁺ and Co²⁺) chemiosmotic efflux pump consists of inner membrane (CzcA), outer membrane (CzcC) and membrane-spanning (CzcB) proteins that together transport cations from the cytoplasm across the periplasmic space to the outside of the cell.     

Probing the coordination properties of glutathione with transition metal ions (Cr2+,Mn2+,Fe2+,Co2+, Ni2+,Cu2+,Zn2+,Cd2+,Hg2+) by density functional theory

Complexes formed by reduced glutathione (GSH) with metal cations (Cr²⁺, Mn²⁺,Fe²⁺,Co²⁺,Ni²⁺,Cu²⁺,Zn²⁺,Cd²⁺,Hg²⁺) were systematically investigated by the density functional theory (DFT). The results showed that the interactions of the metal cations with GSH resulted in nine different stable complexes and many factors had an effect on the binding energy. Generally, for the same period of metal ions, the binding energies ranked in the order of Cu²⁺>Ni²⁺>Co²⁺>Fe²⁺>Cr²⁺>Zn²⁺>Mn²⁺; and for the same group of metal ions, the general trend of binding energies was Zn²⁺>Hg²⁺>Cd²⁺. Moreover, the amounts of charge transferred from S or N to transition metal cations are greater than that of O atoms. For Fe²⁺,Co²⁺,Ni²⁺,Cu²⁺,Zn²⁺,Cd²⁺ and Hg²⁺ complexes, the values of the Wiberg bond indices (WBIs) of M-S (M denotes metal cations) were larger than that of M-N and M-O; for Cr²⁺ complexes, most of the WBIs of M-O in complexes were higher than that of M-S and M-N. Furthermore, the changes in the electron configuration of the metal cations before and after chelate reaction revealed that Cu²⁺, Ni²⁺,Co²⁺ and Hg²⁺ had obvious tendencies to be reduced to Cu⁺,Ni⁺,Co⁺ and Hg⁺ during the coordination process.     

The interaction of zinc, nickel and cadmium with serum albumin and histidine-rich glycoprotein assessed by equilibrium dialysis and immunoadsorbent chromatography

Human serum albumin (HSA) has been shown to bind 2–3 mol of Zn²⁺, Ni²⁺, or Cd²⁺ per mole of protein with apparent dissociation constants (K_d) in the range of 10 μm. Rabbit histidine-rich glycoprotein (HRG) binds 13, 9, and 6 mol of Zn²⁺, Ni²⁺, and Cd²⁺ per mole of protein, respectively, with apparent K_ds also near 10 μm. However, the binding of metals by HRG exhibits positive cooperativity, so that the apparent K_ds may underestimate HRGs true affinity for metal ions. The relative affinities of HSA and HRG for metal ions were found to be Zn²⁺ > Ni²⁺ > Cd²⁺. In addition, histidine (a serum metal chelator) affected the binding of Ni²⁺ by both proteins but not that of Zn²⁺ or Cd²⁺. At physiological concentrations of HSA (250 μm), HRG (2.5 μm), and histidine (100 μm), HRG bound 36% of the Zn²⁺, 9% of the Ni²⁺, and 13% of the Cd²⁺ at a total metal concentration of 25 μm. Under the same conditions HSA held 37% of the Zn²⁺, 14% of the Ni²⁺, and 56% of the Cd²⁺. Thus, HSA appears to have a lower intrinsic affinity for the three metals than HRG but would be expected to bind a higher proportion of these metals in serum. A specific immunoadsorbent column was prepared and used to study the metal binding by HRG in serum directly. Both ⁶⁵Zn²⁺ and ⁶³Ni²⁺ were associated with HRG in aliquots of rabbit serum after incubation with the corresponding metal ion. This evidence indicates that HRG must be considered as a metal binding component of serum.     

Comparison of Physiological Changes in Euglena gracilis During Exposure to Heavy Metals of Heterotrophic and Autotrophic Cells

The effect of different concentrations of Hg²⁺, Cd²⁺, and Pb²⁺ on ultrastructure, growth, respiration, photosynthesis, chlorophyll content, and metal accumulation in Euglena gracilis was examined. The toxicity of the heavy metals was dependent on the culture medium used and whether cells were grown in the dark or under illumination. Hg²⁺ was the most toxic metal, which showed effects at a concentration as low as 1.5 μM; Cd²⁺ showed an intermediate toxicity (effects observed above 50 μM); and Pb²⁺ was almost ineffective up to 1 mM. Cells grown for several weeks in the dark, in the presence of 1.5 μM Hg²⁺ showed a reduced sensitivity to subsequent exposure to Cd²⁺ or Pb²⁺. The Hg²⁺-pretreated cells also presented an enhanced capacity to accumulate other metals. In comparison, light-grown cells showed a greater Cd²⁺ accumulation, but a lower Pb²⁺ uptake than Hg²⁺-pretreated dark-grown cells. Pretreatment of light-grown cells with Hg²⁺ did not enhance the accumulation of Cd²⁺. These results suggest that the capacity to tolerate heavy metals by Euglena may have mechanistic differences when cells are grown in the dark or under illumination.     

Differential action of Hg2+ and Cd2+ on the phycobilisomes and chlorophyll a fluorescence,and photosystem II dependent electron transport in the cyanobacterium Anabaena flos-aquae

The effect of equimolar concentrations of Hg²⁺ and Cd²⁺ on the whole cell absorption spectra, absorption spectra of the extracted phycocyanin (PC) and fluorescence emission spectra of phycobilisomes (PBS) was investigated in the cells of Anabaena flos-aquae. The PC component of the PBS was found to be extremely sensitive to the Hg²⁺ rather than the Cd²⁺ ions. Further, the results showed that Hg²⁺ and Cd²⁺ induced decrease in the rate of Hill activity (H₂O - DCPIP) was partially restored by the electron donor NH₂OH, not by the diphenyl carbazide. Similarly, chlorophyll a fluorescence emission in the presence of metals showed that addition of NH₂OH could effectively reverse the metal induced alterations in the fluorescence emission intensity. These results, together, suggested that Hg²⁺ and Cd²⁺ caused damage to the photosystems (PS) II reaction center. However, a relatively higher stimulation of the chlorophyll a emission at 695 nm with a red shift of 4.0 nm in the presence of Hg²⁺, and Cd²⁺ induced preferential decrease in the emission intensity at 676 nm as compared with the peak at 695 nm were indicative of the differential action of Hg²⁺ and Cd²⁺ on the PS II.     

A simply synthesized biphenyl substituted piperidin‐4‐one for the fluorescence chemosensing of Cd2+

Ion‐induced change in fluorescence is a straight‐forward method for detection of toxic metal ions showing immediate response. Cadmium ions are toxic to the environment. We report in this paper a piperidine‐4‐one‐based fluorescent chemosensor of Cd²⁺ ions, designed and synthesized by a simple method. The compound is characterized using infra‐red (IR) and ¹H–NMR spectral techniques. The chemosensor showed Cd²⁺ ion selectivity and sensitivity in aqueous solution. The stoichiometry and the binding constants were determined using fluorescence spectroscopy. Piperidine‐4‐one shows a 1:1 stoichiometric binding to Cd²⁺. The limit of detection of Cd²⁺ was reported.     

Highly selective ratiometric fluorescent Zn2+ chemosensor based on diarylethene derivative with bi‐8‐carboxamidoquinoline unit

An asymmetrical diarylethene (1O) with a bi‐8‐carboxamidoquinoline unit was synthesized. Its photochromic and fluorescence performances on stimulation with both light and metal ions showed that the diarylethene could serve as a highly selective ratiometric fluorescent chemosensor to detect Zn²⁺ ions based on internal charge transfer and chelation‐enhanced fluorescence processes. The diarylethene could selectively discriminate Zn²⁺ from Cd²⁺ in acetonitrile. Furthermore, Job's plots based on fluorescence titration and electrospray ionization mass spectrometry analysis showed 1 : 1 binding stoichiometry between 1O and Zn²⁺. The binding constant of 1O with Zn²⁺, estimated using the Benesi–Hildebrand method, and the limit of detection were 3.37 × 10⁵ M^–1 and 4.6 × 10^–8 mol/L, respectively. Additionally, the light and metal‐responsive fluorescence behavior of 1O was used successfully to construct a molecular logic circuit with four inputs and one output. Copyright © 2016 John Wiley & Sons, Ltd.