Probing Lanmodulin's Lanthanide Recognition via Sensitized Luminescence Yields a Platform for Quantification of Terbium in Acid Mine Drainage.

Lanmodulin is the first natural, selective macrochelator for f elements-a protein that binds lanthanides with picomolar affinity at 3 EF hands, motifs that instead bind calcium in most other proteins. Here, we use sensitized terbium luminescence to probe the mechanism of lanthanide recognition by this protein as well as to develop a terbium-specific biosensor that can be applied directly in environmental samples. By incorporating tryptophan residues into specific EF hands, we infer the order of metal binding of these three sites. Despite lanmodulin's remarkable lanthanide binding properties, its coordination of approximately two solvent molecules per site (by luminescence lifetime) and metal dissociation kinetics (koff = 0.02-0.05 s-1, by stopped-flow fluorescence) are revealed to be rather ordinary among EF hands; what sets lanmodulin apart is that metal association is nearly diffusion limited (kon ≈ 109 M-1 s-1). Finally, we show that Trp-substituted lanmodulin can quantify 3 ppb (18 nM) terbium directly in acid mine drainage at pH 3.2 in the presence of a 100-fold excess of other rare earths and a 100 000-fold excess of other metals using a plate reader. These studies not only yield insight into lanmodulin's mechanism of lanthanide recognition and the structures of its metal binding sites but also show that this protein's unique combination of affinity and selectivity outperforms synthetic luminescence-based sensors, opening the door to rapid and inexpensive methods for selective sensing of individual lanthanides in the environment and in-line monitoring in industrial operations.

Rare Earth Element Labeling as a Tool for Assuring the Origin of Eggs and Poultry Products.

Laying hens were fed terbium and thulium supplemented feed in order to introduce a distinctive rare earth element pattern that allows discrimination of labeled from unlabeled poultry products. Samples of egg yolk, egg shells, meat, bones, liver, blood, and feces were analyzed using either conventional or laser ablation inductively coupled plasma mass spectrometry. Already after a short time of administering supplemented feed, terbium and thulium enrichment could be unambiguously detected in the products, while absolute terbium and thulium contents remained low enough to ensure safety for the customer. This method could potentially be applied to specifically label foodstuffs produced in certain regions or under certain conditions, in order to ensure food authenticity.

Development of a Click-Chemistry Reagent Compatible with Mass Cytometry.

The recent development of mass cytometry has allowed simultaneous detection of 40 or more unique parameters from individual single cells. While similar to flow cytometry, which is based on detection of fluorophores, one key distinguishing feature of mass cytometry is the detection of atomic masses of lanthanides by mass spectrometry in a mass cytometer. Its superior mass resolution results in lack of signal overlap, thereby allowing multiparametric detection of molecular features in each single cell greater than that of flow cytometry, which is limited to 20 parameters. Unfortunately, most detection in mass cytometry relies on lanthanide-tagged antibodies, which is ideal to detect proteins, but not other types of molecular features. To further expand the repertoire of molecular features that are detectable by mass cytometry, we developed a lanthanide-chelated, azide-containing probe that allows click-chemistry mediated labeling of target molecules. Following incorporation of the thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) during DNA synthesis in S-phase of the cell cycle, we demonstrate that the probe introduced here, tagged with Terbium-159 (159Tb), reacts via copper-catalyzed azide-alkyne Huisgen cycloaddition (click-chemistry) with Edu. Thus, detection of 159Tb makes it possible to measure DNA synthesis in single cells using mass cytometry. The approach introduced here shows similar sensitivity (true positive rate) to other methods used to measure DNA synthesis in single cells by mass cytometry and is compatible with the parallel antibody-based detection of other parameters in single cells. Due to its universal nature, the use of click-chemistry in mass cytometry expands the types of molecular targets that can be monitored by mass cytometry.

Dissolved rare earth elements in the central-western sector of the Ross Sea, Southern Ocean: Geochemical tracing of seawater masses.

The present essay contributes to the existing literature on rare earth elements (REEs) in the southern hemisphere by presenting the first data, to our knowledge, on the vertical profiles of dissolved REEs in 71 samples collected in the central-western sector of the Ross Sea (Southern Ocean-SO). The REEs were measured in the water samples collected during the 2002-2003 and 2005-2006 austral summers. 4 samples were collected and analysed in the framework of a test experiment, as part of the WISSARD Project (Whillans Ice Stream Subglacial Access Research Drilling). Our results show significant differences between the REE patterns of the main water masses present in the SO: we could observe specific signature in the High Salinity Shelf Water (HSSW), Ice Shelf Water (ISW) and Low Salinity Shelf Water (LSSW). A significant increase in Terbium (Tb) concentration was observed in the HSSW and ISW, the two principal water masses contributing to the formation of Antarctic Bottom Water (AABW) in the Ross Sea area, and in LSSW. Some of the HSSW samples show enrichment in Neodymium (Nd). Dissolved REE could therefore be used as tracers to understand the deep circulation of the SO (Pacific sector). We hypothesize that: (I) the characteristic dissolved REE pattern may derive from the composition of source area and from the hydrothermal activity of the central-western area of the Ross Sea; (II) the Tb anomaly observed in the AABW on the South Australian platform could be partially explained by the contribution of AABW generated in the Ross Sea region.

A hydrometallurgical process for the recovery of terbium from fluorescent lamps: Experimental design, optimization of acid leaching process and process analysis.

Terbium and rare earths recovery from fluorescent powders of exhausted lamps by acid leaching with hydrochloric acid was the objective of this study. In order to investigate the factors affecting leaching a series of experiments was performed in according to a full factorial plan with four variables and two levels (42). The factors studied were temperature, concentration of acid, pulp density and leaching time. Experimental conditions of terbium dissolution were optimized by statistical analysis. The results showed that temperature and pulp density were significant with a positive and negative effect, respectively. The empirical mathematical model deducted by experimental data demonstrated that terbium content was completely dissolved under the following conditions: 90 °C, 2 M hydrochloric acid and 5% of pulp density; while when the pulp density was 15% an extraction of 83% could be obtained at 90 °C and 5 M hydrochloric acid. Finally a flow sheet for the recovery of rare earth elements was proposed. The process was tested and simulated by commercial software for the chemical processes. The mass balance of the process was calculated: from 1 ton of initial powder it was possible to obtain around 160 kg of a concentrate of rare earths having a purity of 99%. The main rare earths elements in the final product was yttrium oxide (86.43%) following by cerium oxide (4.11%), lanthanum oxide (3.18%), europium oxide (3.08%) and terbium oxide (2.20%). The estimated total recovery of the rare earths elements was around 70% for yttrium and europium and 80% for the other rare earths.

A Selective Na(+) Aptamer Dissected by Sensitized Tb(3+) Luminescence.

A previous study of two RNA-cleaving DNAzymes, NaA43 and Ce13d, revealed the possibility of a common Na(+) aptamer motif. Because Na(+) binding to DNA is a fundamental biochemical problem, the interaction between Ce13d and Na(+) was studied in detail by using sensitized Tb(3+) luminescence spectroscopy. Na(+) displaces Tb(3+) from the DNAzyme, and thus quenches the emission from Tb(3+) . The overall requirement for Na(+) binding includes the hairpin and the highly conserved 16-nucleotide loop in the enzyme strand, along with a few unpaired nucleotides in the substrate. Mutation studies indicate good correlation between Na(+) binding and cleavage activity, thus suggesting a critical role of Na(+) binding for the enzyme activity. Ce13d displayed a Kd of ∼20 mm with Na(+) (other monovalent cations: 40-60 mm). The Kd values for other metal ions are mainly due to non-specific competition. With a single nucleotide mutation, the specific Na(+) binding was lost. Another mutant improved Kd to 8 mm with Na(+) . This study has demonstrated a Na(+) aptamer with important biological implications and analytical applications. It has also defined the structural requirements for Na(+) binding and produced an improved mutant.

Determining the Effects of Membrane-Interacting Peptides on Membrane Integrity.

In the study of cell-penetrating and membrane-translocating peptides, a fundamental question occurs as to the contribution arising from fundamental peptide-membrane interactions, relative to the contribution arising from the biology and energy of the cell, mostly occurring in the form of endocytosis and subsequent events. A commonly used approach to begin addressing these mechanistic questions is to measure the degree to which peptides can interact with, and physically disrupt, the integrity of synthetic lipid bilayers. Here, we describe a set of experimental methods that can be used to measure the potency, kinetics, transience, and the effective size of peptide-induced membrane disruption.

A label-free fluorescent biosensor for ultratrace detection of terbium (ш) based on structural conversion of G-quadruplex DNA mediated by ThT and terbium (ш).

In this paper, a novel label-free fluorescent biosensor for terbium (ш) (Tb(3+)) was proposed based on structural conversion of G-quadruplex DNA mediated by Thioflavin T (ThT) and Tb(3+). In the presence of K(+), ThT could bind to K(+)-stabilized parallel G-quadruplex, giving rise to high fluorescence intensity. Upon the addition of Tb(3+), Tb(3+) could competitively bind to parallel G-quadruplex leading to the structural change, which resulted in fluorescence decrease. The change of fluorescence intensity (ΔF=F0-F) showed a good linear response toward the concentration of Tb(3+) over the range from 1.0 pM to 10.0 µM with a limit of detection of 0.55 pM. This proposed biosensor was simple and cost-effective in design and in operation with ultrahigh sensitivity and selectivity. Thus, the proposed biosensor could be a promising candidate for monitoring ultratrace Tb(3+) in environment.

Toxic effects of heavy metal terbium ion on the composition and functions of cell membrane in horseradish roots.

The environmental safety of rare earth elements (REEs), especially the toxic effect of REEs on plants, has attracted increasing attention. However, the cellular mechanism of this toxic effect remains largely unknown. Here, the toxic effects of heavy REE terbium ion [Tb(III)] on the cell membrane of horseradish roots were investigated by using electron microscope autoradiography (EMARG) and histochemical methods. The results indicated that Tb(III) was distributed in the extracellular and intracellular spaces of the roots after horseradish was treated with Tb(III). Moreover, the percentage contents of the unsaturated fatty acids in the membrane lipids, the current of the outward K(+) channel and the average diameter of membrane proteins in the roots of horseradish treated with Tb(III) were decreased; on the contrary, the percentage contents of the saturated fatty acids and malondialdehyde in the roots of horseradish treated with Tb(III) were increased. Furthermore, the contents of intracellular N, P, Mg and Fe in the roots of horseradish treated with Tb(III) were decreased, while the contents of intracellular K and Ca in the roots of horseradish treated with Tb(III) were increased. Finally, the effects of Tb(III) on horseradish roots were increased with increasing concentration or duration of Tb(III) treatment. In conclusion, after horseradish was treated with Tb(III), Tb(III) could enter the cells of horseradish roots and lead to the toxic effects on horseradish, which caused the oxidation of the unsaturated fatty acids in the membrane lipids, the changes in the membrane proteins (including the outward K(+) channel), the decrease in the membrane fluidity, and then the inhibition of the intracellular/extracellular-ion exchange in horseradish roots.

A time-resolved luminescence biosensor assay for anaplastic lymphoma kinase (ALK) activity.

A novel time-resolved luminescence biosensor assay for anaplastic lymphoma kinase (ALK) was developed. We used a straightforward strategy to modify a known ALK substrate into a peptide biosensor that can accommodate terbium luminescence sensitization upon its phosphorylation by ALK. Since this strategy is generalizable, this high-throughput screening compatible assay serves as an example for development of other kinase assays that employ terbium luminescence as a read-out.

DNA-based sensitization of Tb3+ luminescence regulated by Ag+ and cysteine: use as a logic gate and a H2O2 sensor.

A simple and facile strategy was developed for regulating the luminescence of Tb(3+) sensitized by DNA, in which Ag(+) and cysteine (Cys) act as activators. The Ag(+)/Cys-mediated reversible luminescence changes in the Tb(3+)-DNA sensing system enabled the design of a DNA INHIBIT logic gate and a H2O2 sensor in a time-resolved luminescence format.

Cytotoxicity and cell imaging potentials of submicron color-tunable yttria particles.

Increased demand of environment protection encouraged scientists to design products and processes that minimize the use and generation of hazardous substances. This work presents comprehensive result of large-scale fabrication and investigation of red-to-green tunable submicron spherical yttria particles codoped with low concentrations of Eu(+3) and Tb(+3). The color emission of synthesized particles can be precisely tuned from red to green by simple variation of Tb/Eu ratio and excitation wavelength. The Tb/Eu-codoped Y(2)O(3) particles did not adversely affect the viability of L-929 fibroblastic cells at concentrations less than 62.5 ppm. Through internalization and wide distribution inside the cells, Tb/Eu codoped Y(2)O(3) particles with intense bright green or red fluorescence rendered cell imaging to be possible. The high brightness, excellent stability, low-toxicity, and imaging capability along with fine color-tunability of synthesized particles enable to find promising application in various areas.

Ce-Tb-Mn co-doped white light emitting glasses suitable for long-wavelength UV excitation.

We report on a new kind of white light emitting glass suitable for long-wavelength ultraviolet excitation by simultaneously emitting blue, green and red fluorescence, which is fabricated by melting of Ce(3+)-Tb(3+)-Mn(2+) co-doped borosilicate glass. The spectroscopic properties of singly, doubly and triply doped glasses have been reported and the energy transfer from Ce(3+) to Tb(3+) and Mn(2+) has also been investigated. By adjusting the concentration of different co-dopants, we obtained the ideal white light emitting borosilicate glass with the color coordinate (x = 0.318, y = 0.333).

A novel Tb(3+)-promoted G-quadruplex-hemin DNAzyme for the development of label-free visual biosensors.

A Tb(3+)-promoted G-quadruplex-hemin DNAzyme was first reported in here. We demonstrated that trace Tb(3+) is able to induce guanine-rich DNA (5'-TGGGTAGGGCGGGTTGGGAAA-3') folding into a compact antiparallel G-quadruplex structure and thus allows the formation of G-quadruplex-hemin DNAzyme. The proposed DNAzyme can effectively catalyze the H(2)O(2)-mediated oxidation of TMB (3,3',5,5'-tetramethylbenzidine sulfate) and leads to a change from colorless to blue in solution color, which provides a sensing platform for the label-free visual detection of Tb(3+). Using above sensing platform, a selective and sensitive label-free visual method for the detection of trace Tb(3+) was developed. The proposed method can be used to detect as low as 1.13×10(-7) M of Tb(3+) by the naked eyes observation and 9.0×10(-9) M of Tb(3+) by UV-vis spectrophotometry with a better stability and reproducibility. Compared with K(+)-promoted G-quadruplex-hemin DNAzyme reported in previous study, the novel Tb(3+)-promoted G-quadruplex-hemin DNAzyme has much higher peroxidase activity and better specificity, which lead to a great potential in the development of optical, electrochemical and chemiluminescence DNAzyme-based biosensors.

Determination of rare earth elements (REES) in airborne particulate matter (APM) collected in Tokyo, Japan, and a positive anomaly of europium and terbium.

The determination of rare earth elements (REEs) in airborne particulate matter (APM) was conducted, and the distribution pattern of atmospheric REEs was evaluated in this study. The APM was collected in the center of Tokyo, Japan, where serious air pollution is always of concern. A cellulose acetate membrane filter was used to collect the APM because Ba and REEs contamination is lower than that in a quartz glass fiber filter. The REEs measurement was conducted by ICP-MS after the digestion of the APM by a microwave acid digestion procedure. The standard reference material (SRM) of NIST 1648 urban particulate matter was used to validate the accuracy of the analytical method. The analytical results for SRM well agreed with those of the reference and reported values. Consequently, the analytical method established in this study was applied to the determination of REEs in APM collected in Tokyo, Japan. The obtained REEs distribution pattern in the APM showed a positive anomaly of Tb and Eu. The La/Sm ratio, which is considered to be as a good indicator of the anthropogenic effect, in size-classified APM showed a high degree of the anthropogenic effect in fine APM with a diameter of <1.1 µm. Emission sources of Tb, Eu and other REEs are discussed.

Use of RNA-bound Tb3+ as a FRET donor.

Lanthanide ions such as Tb(3+) and Eu(3+) have long been used to probe RNA and protein structures due to their luminescence properties and their steric and chemical similarities to biological metal ions such as Mg(2+) and Ca(2+). In this article, we introduce a method that utilizes the enhanced Tb(3+) luminescence upon site-binding to RNA molecules as a FRET donor. Using this method, it is possible to identify specific metal ion-binding locations within a folded RNA molecule. Applications of this method include introducing FRET donors into molecules of interest by engineering RNA loops or bulges as Tb(3+)-binding pockets.

Determination of terbium in phosphate rock by Tb3+-selective fluorimetric optode based on dansyl derivative as a neutral fluorogenic ionophore.

For the first time a highly sensitive and selective fluorimetric optode membrane was prepared for determination of trace amounts of Tb(III) ions in phosphate rock samples. The Tb(III) sensing system was constructed by incorporating 5-(dimethylamino)-N'-(2-hydroxy-1-naphthoyl) naphthalene-1-sulfonohydrazine (L) as a neutral Tb(III)-selective fluoroionophore, in the plasticized PVC membrane containing sodium tetraphenyl borate as a liphophilic anionic additive. The response of the optode is based on the strong fluorescence quenching of L by Tb(3+) ions. At a pH value of 5.0, the optode displays a wide concentration range of 1.0 x 10(-7) to 1.0 x 10(-2)M, with a relatively fast response time of less than 45 s. In addition, to high stability and reproducibility, the sensor shows a unique selectivity towards Tb(3+) ion with respect to common cations. The optode was applied successfully to the trace determination of terbium ion in binary mixture and water samples and the determination of Tb(3+) in phosphate rock samples.

An electrochemical biosensor for ultratrace terbium based on Tb3+ promoted conformational change of human telomeric G-quadruplex.

A new electrochemical biosensor for the monitoring of ultratrace terbium based on the conformational change of DNA containing a single guanine (G)-rich stretch was described here. The biosensor was fabricated by immobilizing a thiolated DNA containing a single G-rich stretch on the gold surface as probe surface. The G-rich DNA probe was found to be capable of changing its configuration from flexible single-stranded structures to rigid tetramolecular G-quadruplex in the presence of terbium III, which provided a switchable charge transport path for the oxidation of [Fe(CN)(6)](4-). The switchable surface provided a sensing platform for the single-step and reagentless detection of Tb(3+). Using this reusable electrochemical sensing platform, a simple, rapid, and selective biosensor for the determination of ultratrace terbium ions with a detection limit of 6.0 x 10(-11)M has been developed. The success in the present biosensor served as a significant step toward the development of monitoring ultratrace Tb(3+) in river water or seawater.

A new Tb3+-selective fluorescent sensor based on 2-(5-(dimethylamino)naphthalen-1-ylsulfonyl)-N-henylhydrazinecarbothioamide.

A novel fluorescent chemosensor 2-(5-(dimethylamino)naphthalen-1-ylsulfonyl)-N-phenylhydrazinecarbothioamide (L) has been synthesized, which revealed an emission of 530 nm and when excited at 360 nm. The fluorescent probe undergoes a fluorescent emission intensity quenching upon binding to terbium ions in MeCN solution. The fluorescence quenching of L is attributed to the 1:1 complex formation between L and Tb(III) which has been utilized as the basis for the selective detection of Tb(III). The linear response range covers a concentration range of Tb(III) from 4.0 x 10(-7) to 1.0 x 10(-5) M and the detection limit is 1.4 x 10(-7) M. The association constant of the 1:1 complex formation for L-Tb(+3) was calculated to be 6.01 x 10(6) M(-1), and the fluorescent probe exhibits high selectivity over other common metal ions mono-, di-, and trivalent cations indicate good selectivity for Tb(III) ions over a large number of interfering cations.

Fluorescence lifetime and intensity of terbium-doped dipicolinic acid in water, HCl, and sodium acetate buffer solutions.

The lifetimes of the individual fluorescing lines from the terbium-doped dipicolinic acid (DPA) complex have been measured and reported, for the first time to our knowledge. These lifetimes have been measured as a function of terbium and dipicolinic acid concentration, solvent pH, and solvent composition for water, HCl, and sodium acetate buffer solutions. Fluorescence lifetimes over the range from 0.75 to 1.07 ms were measured. The maximum fluorescence was obtained for distilled water solutions.