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Thallium (Tl) is a toxic element associated with minerals, and its redistribution is facilitated by both geological and anthropogenic activities. In the natural environment, the transformation and migration of Tl mediated by (micro)organisms have attracted increasing attention. This review presents an overview of the biochemical transformation of Tl and the bioremediation strategies for Tl contamination. In the environment, Tl exists in various forms and originates from diverse sources. The global distribution characteristics of Tl in various media are summarized here, while its speciation and toxicity mechanism to organisms are elucidated. Interactions between (micro)organisms and Tl are commonly observed in the environment. Microbial response mechanisms to typical Tl exposure are analyzed at both species and gene levels, and the possibility of microorganisms as bio-indicators for monitoring Tl contamination is also highlighted. The processes and mechanisms involved in the microbial and benthic mediated transformation of Tl, as well as its enrichment by plants, are discussed. Additionally, in situ bioremediation strategies for Tl contamination and bio-treatment techniques for Tl-containing wastewater are summarized. Finally, the existing knowledge gaps and future research challenges are emphasized, including Tl distribution characteristics in the atmosphere and ocean, the key molecular mechanisms underlying Tl transformation by organisms, the screening of potential Tl oxidizing microorganisms and hyperaccumulators, as well as the revelation of global biogeochemical cycling pathways of Tl., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

To reduce heavy metal toxicity, like that induced by thallium (TI) in plants, growth-promoting bacteria (GPB) are a widely used to enhance plant tolerance to heavy metals toxicity. In our study, we characterized seven GPB and identified Actinoplanes spp., as the most active strain. This bioactive strain was then applied to alleviate TI phytotoxicity. TI contamination (20 mg/kg soil) induced TI bioaccumulation, reducing wheat growth (biomass accumulation) and photosynthesis rate, by about 55% and 90%, respectively. TI stress also induced oxidative damages as indicated by increased oxidative markers (H 2 O 2 and lipid peroxidation (MDA)). Interestingly, Actinoplanes spp. significantly reduced growth inhibition and oxidative stress by 20% and 70%, respectively. As a defense mechanism to mitigate the TI toxicity, wheat plants showed improved antioxidant and detoxification defense including increased phenolic and tocopherols levels as well as peroxidase (POX), catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GR) enzymes activities. These defense mechanisms were further induced by Actinoplanes spp. Additionally, Actinoplanes spp. increased the production of heavy metal-binding ligands such as metallothionein, phytochelatins, total glutathione, and glutathione S-transferase activity by 100%, 90%, 120%, and 100%, respectively. This study, therefore, elucidated the physiological and biochemical bases underlying TI-stress mitigation impact of Actinoplanes spp. Overall, Actinoplanes spp. holds promise as a valuable approach for ameliorating TI toxicity in plants. KEYBOARD: Actinobacteria, Bioaccumulation, Detoxification, Membrane damage, Redox regulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Cyclobutane pyrimidine dimer (CPD) and (6-4)photoproduct (6-4 PP) are two major types of UV-induced DNA lesion and 6-4 PP is more mutagenic than CPD. Activated by lesion detection, nucleotide excision repair (NER) eliminates CPDs and 6-4 PPs. Thallium (Tl) is a toxic metal existing primarily as Tl + in the aquatic environment. Ingestion of Tl + -contaminated foods and water is a major route of human poisoning. As Tl + may inhibit enzyme activities via binding to sulfhydryl groups, this study explored if Tl + could intensify UV mutagenicity by inactivating NER-linked damage recognition factors using zebrafish (Danio rerio) embryo as a model system. Incubation of Tl + (as thallium nitrate) at 0.1-0.4 μg/mL with zebrafish extracts for 20 min caused a concentration-dependent inhibition of 6-4 PP binding activities as shown by a photolesion-specific band shift assay, while CPD binding activities were insensitive to Tl + . The ability of Tl + to suppress 6-4 PP detection was stronger than that of Hg 2+ . Exposure of zebrafish embryos at 1 h post fertilization (hpf) to Tl + at 0.4-1 μg/mL for 9 or 71 h also specifically inhibited 6-4 PP detection, indicating that Tl + induced a prolonged inhibition of 6-4 PP sensing ability primarily via its direct interaction with damage recognition molecules. Tl + -mediated inhibition of 6-4 PP binding in embryos at distinct stages resulted in a suppression of NER capacity monitored by a transcription-based DNA repair assay. Our results revealed the potential of Tl + to enhance UV mutagenicity by disturbing the removal of 6-4 PP through repressing the lesion detection step of NER., Competing Interests: Declaration of competing interest The authors declare that there are no competing interests that could have appeared to influence the work reported in this article., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The development of cell-based fluorescent assays has resulted in an incredible tool for searching new ion channels' modulators with a biophysical and clinical profile. Among all the ion channels, potassium (K + )-permeable channels represent the most diverse and relevant for cell function, making them attractive targets for drug discovery. Some of the cell-based assays for K + channels take advantage of a thallium-sensitive dye whose fluorescence increased upon the binding of thallium (Tl + ), an ion able to move through K + channels. We optimize the FLIPR Potassium Assay Kit based on thallium influx to measure the Kv10.1 activity., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Thallium (Tl), a key element in high-tech industries, is recognized as a priority pollutant by the US EPA and EC. Tl accumulation threatens aquatic ecosystems. Despite its toxicity, little is known about its impact on cyanobacteria. This study explores the biochemical mechanisms of Tl(I) toxicity in cyanobacteria, focusing on physiology, metabolism, oxidative damage, and antioxidant responses. To this end, Anabaena and Nostoc were exposed to 400 µg/L, and 800 µg/L of Tl(I) over seven days. Anabaena showed superior Tl(I) accumulation with 7.8% removal at 400 µg/L and 9.5% at 800 µg/L, while Nostoc removed 2.2% and 7.4%, respectively. Tl(I) exposure significantly reduced the photosynthesis rate and function, more than in Nostoc . It also altered primary metabolism, increasing sugar levels and led to higher amino and fatty acids levels. While Tl(I) induced cellular damage in both species, Anabaena was less affected. Both species enhanced their antioxidant defense systems, with Anabaena showing a 175.6% increase in SOD levels under a high Tl(I) dose. This suggests that Anabaena's robust biosorption and antioxidant systems could be effective for Tl(I) removal. The study improves our understanding of Tl(I) toxicity, tolerance, and phycoremediation in cyanobacteria, aiding future bioremediation strategies.

Thallium (Tl) is a priority pollutant regulated by the US EPA. It is also a critical element commonly used in high technology industries; with an increasing demand for semiconductors nowadays, wastewater discharges from manufacturing plants or metal mining activities may result in elevated levels of thallium in receiving water harming aquatic organisms. Regarding the impact of thallium on freshwater algae, little attention has been paid to prokaryotic physiology through various exposure periods. In this bench-scale study, prokaryotic alga Microcystis aeruginosa PCC 7806 was cultured in modified BG11 medium and exposed to Tl + (TlNO 3 ) ranging from 250 to 1250 μg/L for 4 and 14 days. Throughout the experiment using flow cytometry assays, algal population, cell membrane integrity, oxidation stress level, and chlorophyll fluorescence were exacerbated following the exposure to 750 μg Tl/L (approximately 4-day effective concentration of Tl + for reducing 50% of algal population). Potassium and humic acid (HA) (1-5 mg/L) were added to study their influences on the thallium toxicity. With the additions of potassium, thallium toxicities to algal population and physiology were not significantly changed within 4 days, while they were alleviated within 14 days. With the addition of HA at 1 mg/L, cell membrane integrity was significantly attenuated within 4 days; ameliorating effects on algal population and oxidative stress were not observed until day 14. Thallium toxicities on oxidative stress level and photosynthesis activity were exacerbated in the presence of HA at 3-5 mg/L. The study provides useful information for further studies on the mode of toxic action of Tl + in prokaryotic algae; it also demonstrates the necessity of considering short and long-term exposure durations while incorporating water chemistry into assessment of thallium toxicity to algae., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Kuo-Pei Tsai reports financial support was provided by The Ministry of Science and Technology in Taiwan. Kuo-Pei Tsai reports financial support was provided by The Ministry of Education in Taiwan., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Malignant migrating partial seizure of infancy (MMPSI) is a devastating and pharmacoresistant form of infantile epilepsy. MMPSI has been linked to multiple gain-of-function (GOF) mutations in the KCNT1 gene, which encodes for a potassium channel often referred to as SLACK. SLACK channels are sodium-activated potassium channels distributed throughout the central nervous system (CNS) and the periphery. The investigation described here aims to discover SLACK channel inhibitor tool compounds and profile their pharmacokinetic and pharmacodynamic properties. A SLACK channel inhibitor VU0531245 (VU245) was identified via a high-throughput screen (HTS) campaign. Structure-activity relationship (SAR) studies were conducted in five distinct regions of the hit VU245. VU245 analogs were evaluated for their ability to affect SLACK channel activity using a thallium flux assay in HEK-293 cells stably expressing wild-type (WT) human SLACK. Selected analogs were tested for metabolic stability in mouse liver microsomes and plasma-protein binding in mouse plasma. The same set of analogs was tested via thallium flux for activity versus human A934T SLACK and other structurally related potassium channels, including SLICK and Maxi-K. In addition, potencies for selected VU245 analogs were obtained using whole-cell electrophysiology (EP) assays in CHO cells stably expressing WT human SLACK through an automated patch clamp system. Results revealed that this scaffold tolerates structural changes in some regions, with some analogs demonstrating improved SLACK inhibitory activity, good selectivity against the other channels tested, and modest improvements in metabolic clearance. Analog VU0935685 represents a new, structurally distinct small-molecule inhibitor of SLACK channels that can serve as an in vitro tool for studying this target., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: C. David Weaver, co-author of this manuscript, is an owner of WaveFront Biosciences and ION biosciences, makers of the Panoptic plate reader and Thallos, Tl(+)-sensitive fluorescent indicators used in the these studies, respectively., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The similarities between thallium and potassium have suggested the use of calcium polystyrene sulfonate (CPS), an oral ion exchange resin, as a potential agent against thallium intoxication. Therefore, the study was an attempt to evaluate the efficacy of CPS and Prussian blue when given alone or in combination against thallium toxicity. The effect on binding capacity was investigated in terms of contact time, amount of CPS, influence of pH, simulated physiological solutions and interference of potassium ions. Also, rats were given single dose of thallium chloride (20 mg kg -1 ) and the treatment with PB and CPS was given for 28 days as CPS 30 g kg -1 , orally, twice a day, PB 3 g kg -1 , orally, twice a day and their combination. The effect of antidotal treatment was evaluated by calculating the thallium levels in various organs, blood, urine and feces. The results of the in vitro study indicated exceedingly quick binding in the combination of CPS and PB as compared to PB alone. Also, it was found that the binding capacity at pH 2.0 was considerably increased for PB with CPS (184.656 mg g -1 ) as compared to PB (37.771 mg g -1 ). Furthermore, statistically significant results were obtained in the in vivo study as after 7th day, thallium levels in blood of rats treated with combination were reduced by 64% as compared to control group and 52% as compared to alone PB treated group. Also, Tl retention in liver, kidney, stomach, colon and small intestine of combination treated rats was significantly reduced to 46%, 28%, 41%, 32% and 33% respectively, as compared to alone PB treated group. These findings demonstrate this as a good antidotal option against thallium intoxication., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Thallium (Tl + ) is a trace metal with extreme toxicity and is highly soluble in water, posing a great risk to ecological and human safety. This work aimed to investigate the role played by Tl + in regulating lipid accumulation in microalgae and the removal efficiency of Tl + . The effect of Tl + on the cell growth, lipid production and Tl + removal efficiency of Parachlorella kessleri R-3 was studied. Low concentrations of Tl + had no significant effect on the biomass of microalgae. When the Tl + concentration exceeded 5 μg L -1 , the biomass of microalgae showed significant decrease. The highest lipid content of 63.65% and lipid productivity of 334.55 mg L -1 d -1 were obtained in microalgae treated with 10 and 5 μg L -1 Tl + , respectively. Microalgae can efficiently remove Tl + and the Tl + removal efficiency can reach 100% at Tl + concentrations of 0-25 μg L -1 . The maximum nitric oxide (NO) level of 470.48 fluorescence intensity (1 × 10 6 cells) -1 and glutathione (GSH) content of 343.51 nmol g -1 (fresh alga) were obtained under 5 μg L -1 Tl + stress conditions. Furthermore, the exogenous donor sodium nitroprusside (SNP) supplemented with NO was induced in microalgae to obtain a high lipid content (59.99%), lipid productivity (397.99 mg L -1 d -1 ) and GSH content (430.22 nmol g -1 (fresh alga)). The corresponding analysis results indicated that NO could participate in the signal transduction pathway through modulation of reactive oxygen species (ROS) signaling to activate the antioxidant system by increasing the GSH content to eliminate oxidative damage induced by Tl + stress. In addition, NO regulation of ROS signaling may enhance transcription factors associated with lipid synthesis, which stimulates the expression of genes related to lipid synthesis, leading to increased lipid biosynthesis in microalgae. Moreover, it was found that the change in Tl + had little effect on the fatty acid components and biodiesel properties. This study showed that Tl + stress can promote lipid accumulation in microalgae for biodiesel production and simultaneously effectively remove Tl + , which provided evidence that NO was involved in signal transduction and antioxidant defense, and improved the understanding of the interrelation between NO and ROS to regulate lipid accumulation in microalgae., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Thallium (Tl) is a rare trace metal element but increasingly detected in wastewater produced by coal-burning, smelting, and more recently, high-tech manufacturing industries. However, the adverse effects of Tl, especially cardiotoxicity, on aquatic biota remain unclear. In this study, zebrafish model was used to elucidate the effects and mechanisms of Tl(I) cardiotoxicity in developing embryos. Exposure of embryonic zebrafish to low-dose Tl(I) (25-100 μg/L) decreased heart rate and blood flow activity, and subsequently impaired swim bladder inflation and locomotive behavior of larvae. Following high-level Tl(I) administration (200-800 μg/L), embryonic zebrafish exhibited pericardial edema, incorrect heart looping, and thinner myocardial layer. Based on RNA-sequencing, Tl(I) altered pathways responsible for protein folding and transmembrane transport, as well as negative regulation of heart rate and cardiac jelly development. The gene expression of nppa, nppb, ucp1, and ucp3, biomarkers of cardiac damage, were significantly upregulated by Tl(I). Our findings demonstrate that Tl(I) at environmentally relevant concentrations interfered with cardiac development with respect to anatomy, function, and transcriptomic alterations. The cardiotoxic mechanisms of Tl(I) provide valuable information in the assessment of Tl-related ecological risk in freshwater environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Although several studies have evaluated the effects of Thallium (Tl) in adult species of fish, the developmental toxicity of Tl has not been previously explored. In this study, zebrafish embryos (<4 h post fertilization (hpf)) were exposed to Tl at concentrations from 0.8 to 400 μg L -1 for 7 d. The results showed that the decreased hatching rate and increased malformation rate were observed in the larvae. The swimming velocity of larvae from 200 and 400 μg L -1 treatments was respectively reduced by ~26 % and 15 %. Histopathological analysis of liver indicated the number of cells of karyolysis (143 % and 202 %) and pyknosis (170 % and 131 %) were respectively increased in 200 and 400 μg L -1 Tl treatments. Meanwhile, the Tl body burden and metallothionein (MT) levels in the larvae were increased with elevated Tl concentrations. The level of malondialdehyde (MDA) was increased by ~20 to 51 % in all Tl treatments and total antioxidant capacity (TAC) was decreased by ~12 % at 200 μg L -1 . The activities of Na + /K + -ATPase and protease were inhibited in 200 and 400 μg L -1 Tl treatments. Moreover, the transcripts of genes (Nrf2, HO-1, TNF-α, IL-1β, IL-8, IL-10, TGF) were significantly altered. In addition, a total of 930 differentially expressed genes (DEGs) and 1549 DEGs were found in the 200 and 400 μg L -1 treatments with 458 overlapped DEGs by transcriptomic analysis. The protein digestion and absorption, ECM-receptor interaction, and complement and coagulation cascades pathways were shown to be the most significantly enriched pathways. This study helps better understand the molecular mechanisms of Tl toxicity in fish., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Thallium (Tl), a highly toxic and priority pollutant heavy metal, exposure can damage mitochondria and disrupt their function. The liver is the central organ that controls lipid homeostasis and contains a large number of mitochondria. So far, there is no study investigating the effects of Tl exposure on hepatic fatty acid metabolism. Here, we showed that 10 ppm of Tl(I) and Tl(III) exposures for two weeks did not significantly affect the body weight and water/food intake in mice. However, it decreased the ratio of liver/weight and induced hepatic sinus congestion and hepatocyte necrosis. Inductively coupled plasma-mass spectrometry (ICP-MS) analysis revealed Tl accumulation in the liver. Gas chromatography-mass spectrometry (GC-MS) results showed that Tl(I) exposure significantly increased hepatic C18:0 concentration, while significantly decreased the concentrations of C16:1n-7, C20:1n-9, C18:3n-6, and C20:2n-9. Tl(III) exposure significantly reduced hepatic concentrations of C20:0, C22:0, C20:1n-9, C18:3n-6, and C20:3n-6. In addition, Tl(I) exposure upregulated the genes related to antioxidation (HO-1, GPX1, and GPX4), fatty acid synthesis (FADS2 and Elovl2), and fatty acid oxidation pathway (PPARα, ACADM, ACADVL, ACAA2, and CPT1A) in the liver. Tl(III) exposure did not significantly affect the transcript levels of liver antioxidative/metabolic enzymes and fatty acid synthesis-related genes, but upregulated fatty acid oxidation pathway-related genes (CYP4A10 and CPT1A). These results suggest that Tl(I) and Tl(III) exposures can cause liver damage and disrupt hepatic fatty acid metabolism, which provide new insights into Tl exposure-induced energy depletion from the perspective of fatty acid metabolism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Experimental liver injury with hepatocelluar necrosis and abnormal liver tests is caused by exposure to heavy metals (HMs) like aluminum, arsenic, beryllium, cadmium, chromium, cobalt, copper, iron, lead, mercury, molybdenum, nickel, platinum, thallium, titanium, vanadium, and zinc. As pollutants, HMs disturb the ecosystem, and as these substances are toxic, they may affect the health of humans and animals. HMs are not biodegradable and may be deposited preferentially in the liver. The use of animal models can help identify molecular and mechanistic steps leading to the injury. HMs commonly initiate hepatocellular overproduction of ROS (reactive oxygen species) due to oxidative stress, resulting in covalent binding of radicals to macromolecular proteins or lipids existing in membranes of subcellular organelles. Liver injury is facilitated by iron via the Fenton reaction, providing ROS, and is triggered if protective antioxidant systems are exhausted. Ferroptosis syn pyroptosis was recently introduced as mechanistic concept in explanations of nickel (Ni) liver injury. NiCl 2 causes increased iron deposition in the liver, upregulation of cyclooxygenase 2 (COX-2) protein and mRNA expression levels, downregulation of glutathione eroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), nuclear receptor coactivator 4 (NCOA4) protein, and mRNA expression levels. Nickel may cause hepatic injury through mitochondrial damage and ferroptosis, defined as mechanism of iron-dependent cell death, similar to glutamate-induced excitotoxicity but likely distinct from apoptosis, necrosis, and autophagy. Under discussion were additional mechanistic concepts of hepatocellular uptake and biliary excretion of mercury in exposed animals. For instance, the organic anion transporter 3 (Oat3) and the multidrug resistance-associated protein 2 (Mrp2) were involved in the hepatic handling of mercury. Mercury treatment modified the expression of Mrp2 and Oat3 as assessed by immunoblotting, partially explaining its impaired biliary excretion. Concomitantly, a decrease in Oat3 abundance in the hepatocyte plasma membranes was observed that limits the hepatic uptake of mercury ions. Most importantly and shown for the first time in liver injury caused by HMs, titanium changed the diversity of gut microbiota and modified their metabolic functions, leading to increased generation of lipopolysaccharides (LPS). As endotoxins, LPS may trigger and perpetuate the liver injury at the level of gut-liver. In sum, mechanistic and molecular steps of experimental liver injury due to HM administration are complex, with ROS as the key promotional compound. However, additional concepts such as iron used in the Fenton reaction, ferroptosis, modification of transporter systems, and endotoxins derived from diversity of intestinal bacteria at the gut-liver level merit further consideration.

Exposure to metals increases risk for pregnancy complications. Extracellular vesicle (EV) miRNA contribute to maternal-foetal communication and are dysregulated in pregnancy complications. However, metal impacts on maternal circulating EV miRNA during pregnancy are unknown. Our objective was to investigate the impact of multiple metal exposures on EV miRNA in maternal circulation during pregnancy in the MADRES Study. Associations between urinary concentrations of nine metals and 106 EV miRNA in maternal plasma during pregnancy were investigated using robust linear regression (N = 231). Primary analyses focused on metal-miRNA associations in early pregnancy (median: 12.3 weeks gestation). In secondary analyses, we investigated associations with late pregnancy miRNA counts (median: 31.8 weeks gestation) in a subset of participants (N = 184) with paired measures. MiRNA associated with three or more metals (P FDR <0.05) were further investigated using Bayesian Kernel Machine Regression (BKMR), an environmental mixture method. Thirty-five miRNA were associated (P FDR <0.05) with at least one metal in early pregnancy. One association (an inverse association between cobalt and miR-150-5p) remained statistically significant when evaluating late pregnancy miRNA counts. Eight miRNA (miR-302b-3p, miR-199a-5p, miR-188-5p, miR-138-5p, miR-212-3p, miR-608, miR-1272, miR-19b-3p) were associated with three metals (barium, mercury, and thallium) in early pregnancy, and their predicted target genes were enriched in pathways important for placental development. Results were consistent when using BKMR. Early pregnancy exposure to barium, mercury, and thallium may have short-term impacts on a common set of EV miRNA which target pathways important for placental development.

Loss-of-function (LOF) variants in the K V 11.1 potassium channel cause long QT syndrome (LQTS). Most variants disrupt intracellular channel transport (trafficking) to the cell membrane. Since some channel inhibitors improve trafficking of K V 11.1 variants, a high-throughput screening (HTS) assay to detect trafficking enhancement would be valuable to the identification of drug candidates. The thallium (Tl + ) flux assay technique, widely used for drug screening, was optimized using human embryonic kidney (HEK-293) cells expressing a trafficking-deficient K V 11.1 variant in 384-well plates. Assay quality was assessed using Z prime (Z') scores comparing vehicle to E-4031, a drug that increases K V 11.1 membrane trafficking. The optimized assay was validated by immunoblot, electrophysiology experiments, and a pilot drug screen. The combination of: 1) truncating the trafficking-deficient variant K V 11.1-G601S (K V 11.1-G601S-G965*X) with the addition of 2) K V 11.1 channel activator (VU0405601) and 3) cesium (Cs + ) to the Tl + flux assay buffer resulted in an outstanding Z' of 0.83. To validate the optimized trafficking assay, we carried out a pilot screen that identified three drugs (ibutilide, azaperone, and azelastine) that increase K V 11.1 trafficking. The new assay exhibited 100% sensitivity and specificity. Immunoblot and voltage-clamp experiments confirmed that all three drugs identified by the new assay improved membrane trafficking of two additional LQTS K V 11.1 variants. We report two new ways to increase target-specific activity in trafficking assays-genetic modification and channel activation-that yielded a novel HTS assay for identifying drugs that improve membrane expression of pathogenic K V 11.1 variants. SIGNIFICANCE STATEMENT: This manuscript reports the development of a high-throughput assay (thallium flux) to identify drugs that can increase function in K V 11.1 variants that are trafficking-deficient. Two key aspects that improved the resolving power of the assay and could be transferable to other ion channel trafficking-related assays include genetic modification and channel activation., (Copyright © 2022 by The American Society for Pharmacology and Experimental Therapeutics.)

TRAAK (KCNK4, K2P4.1) is a mechanosensitive two-pore domain potassium (K2P) channel. Due to its expression within sensory neurons and genetic link to neuropathic pain it represents a promising potential target for novel analgesics. In common with many other channels in the wider K2P sub-family, there remains a paucity of small molecule pharmacological tools. Specifically, there is a lack of molecules selective for TRAAK over the other members of the TREK subfamily of K2P channels. We developed a thallium flux assay to allow high throughput screening of compounds and facilitate the identification of novel TRAAK activators. Using a library of ∼1200 drug like molecules we identified Aprepitant as a small molecule activator of TRAAK. Aprepitant is an NK-1 antagonist used to treat nausea and vomiting. Close structural analogues of Aprepitant and a range of NK-1 antagonists were also selected or designed for purchase or brief chemical synthesis and screened for their ability to activate TRAAK. Electrophysiology experiments confirmed that Aprepitant activates both the 'long' and 'short' transcript variants of TRAAK. We also demonstrated that Aprepitant is selective and does not activate other members of the K2P superfamily. This work describes the development of a high throughput assay to identify potential TRAAK activators and subsequent identification and confirmation of the novel TRAAK activator Aprepitant. This discovery identifies a useful tool compound which can be used to further probe the function of TRAAK K2P channels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Ion channels have been proposed as therapeutic targets for different types of malignancies. One of the most studied ion channels in cancer is the voltage-gated potassium channel ether-à-go-go 1 or Kv10.1. Various studies have shown that Kv10.1 expression induces the proliferation of several cancer cell lines and in vivo tumor models, while blocking or silencing inhibits proliferation. Kv10.1 is a promising target for drug discovery modulators that could be used in cancer treatment. This work aimed to screen for new Kv10.1 channel modulators using a thallium influx-based assay., Methods: Pharmacological effects of small molecules on Kv10.1 channel activity were studied using a thallium-based fluorescent assay and patch-clamp electrophysiological recordings, both performed in HEK293 stably expressing the human Kv10.1 potassium channel., Results: In thallium-sensitive fluorescent assays, we found that the small molecules loperamide and amitriptyline exert a potent inhibition on the activity of the oncogenic potassium channel Kv10.1. These results were confirmed by electrophysiological recordings, which showed that loperamide and amitriptyline decreased the amplitude of Kv10.1 currents in a dose-dependent manner. Both drugs could be promising tools for further studies., Conclusions: Thallium-sensitive fluorescent assay represents a reliable methodological tool for the primary screening of different molecules with potential activity on Kv10.1 channels or other K + channels., (© 2021. Maj Institute of Pharmacology Polish Academy of Sciences.)

Thallium (TI) is one of the most toxic heavy metals. Human exposure to Tl occurs through contaminated drinking water and from there to food, a threat to health. Recently, environmental contamination by Tl has been reported in several countries, urging the need for studies to determine the impact of endogenous and exogenous mechanisms preventing thallium toxicity. The cytoprotective effect of metallothionein (MT), a protein with high capacity to chelate metals, at two doses (100 and 600 µg/rat), was tested. Prussian blue (PB) (50 mg/kg) was administered alone or in combination with MT. A dose of Tl (16mg/kg) was injected i.p. to Wistar rats. Antidotes were administered twice daily, starting 24h after Tl injection, for 4 days. Tl concentrations diminished in most organs (p < 0.05) by effect of PB, alone or in combination with MT, whereas MT alone decreased Tl concentrations in testis, spleen, lung and liver. Likewise, brain thallium also diminished (p < 0.05) by effect of PB and MT alone or in combination in most of the regions analyzed (p < 0.05). The greatest diminution of Tl was achieved when the antidotes were combined. Plasma markers of renal damage increased after Tl administration, while PB and MT, either alone or in combination, prevented the raise of those markers. Only MT increased the levels of reduced glutathione (GSH) in the kidney. Finally, increased Nrf2 was observed in liver and kidney, after treatment with MT alone or in combination with PB. Results showed that MT alone or in combination with PB is cytoprotective after thallium exposure., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The effects of both Tl + and thiol reagents were studied on the content of the inner membrane free SH-groups, detected with Ellman reagent, and the inner membrane potential as well as swelling and respiration of succinate-energized rat liver mitochondria in medium containing TlNO 3 and KNO 3 . These effects resulted in a rise in swelling and a decrease in the content, the potential, and mitochondrial respiration in 3 and 2,4-dinitrophenol-uncoupled states. A maximal effect was seen when phenylarsine oxide reacting with thiol groups recessed into the hydrophobic regions of the membrane. Compared with phenylarsine oxide, the effective concentrations of other reagents were approximately one order of magnitude higher in experiments with mersalyl and 4,4'-diisothiocyanostilbene-2,2'-disulfonate, and two orders of magnitude higher in experiments with tert-butyl hydroperoxide and diamide. The above effects of Tl + and the thiol reagents became even more pronounced with calcium overload of mitochondria. However, the effects were suppressed by inhibitors of the mitochondrial permeability transition pore (cyclosporine A, ADP, and n-ethylmaleimide). These findings suggest that opening of the pore induced by Tl + in the inner membrane can be dependent on the conformation state of the adenine nucleotide translocase, which depends on the activity of its thiol groups., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Inhibition of the K Ca 3.1 potassium channel has therapeutic potential in a variety of human diseases, including inflammation-associated disorders and cancers. However, K Ca 3.1 inhibitors with high therapeutic promise are currently not available. This study aimed to establish a screening assay for identifying inhibitors of K Ca 3.1 in native cells and from library compounds derived from natural products in Thailand. The screening platform was successfully developed based on a thallium flux assay in intestinal epithelial (T84) cells with a Z' factor of 0.52. The screening of 1352 compounds and functional validation using electrophysiological analyses identified 8 compounds as novel K Ca 3.1 inhibitors with IC 50 values ranging from 0.14 to 6.57 µM. These results indicate that the assay developed is of excellent quality for high-throughput screening and capable of identifying K Ca 3.1 inhibitors. This assay may be useful in identifying novel K Ca 3.1 inhibitors that may have therapeutic potential for inflammation-associated disorders and cancers.

Two-pore domain potassium (K2P) channels carry background (or leak) potassium current and play a key role in regulating resting membrane potential and cellular excitability. Accumulating evidence points to a role for K2Ps in human pathophysiologies, most notably in pain and migraine, making them attractive targets for therapeutic intervention. However, there remains a lack of selective pharmacological tools. The aim of this work was to apply a "target class" approach to investigate the K2P superfamily and identify novel activators across all the described subclasses of K2P channels. Target class drug discovery allows for the leveraging of accumulated knowledge and maximizing synergies across a family of targets and serves as an additional approach to standard target-based screening. A common assay platform using baculovirus (BacMam) to transiently express K2P channels in mammalian cells and a thallium flux assay to determine channel activity was developed, allowing the simultaneous screening of multiple targets. Importantly, this system, by allowing precise titration of channel function, allows optimization to facilitate the identification of activators. A representative set of channels (THIK-1, TWIK-1, TREK-2, TASK-3, and TASK-2) were screened against a library of Food and Drug Administration (FDA)-approved compounds and the LifeArc Index Set. Activators were then analyzed in concentration-response format across all channels to assess selectivity. Using the target class approach to investigate the K2P channels has enabled us to determine which of the K2Ps are amenable to small-molecule activation, de-risk multiple channels from a technical point of view, and identify a diverse range of previously undescribed pharmacology.

Thallium (Tl), a ubiquitous environmental toxicant, can cross the placental barrier during pregnancy. However, the effects of prenatal Tl exposure on placental function are currently unclear. Based on the Ma'anshan Birth Cohort study, we examined whether long-term prenatal Tl exposure was associated with placental inflammation. Tl concentrations were quantified in serum samples (n = 7050) from 2515 pregnancy during each trimester, placental inflammatory cytokine mRNA expression was assessed in 2519 placenta tissues. Geometric mean values of serum Tl concentrations were 63.57, 63.63 and 48.71 ng/L for the first, second and third trimesters, respectively. After adjustment for potential confounders, serum Tl concentration was positively associated with CD68 (β: 0.30; 95% CI: 0.05, 0.56) in the first trimester and TNF-α (β: 0.12; 95% CI: 0.01, 0.23), IL-6 (β: 0.15; 95% CI: 0.05, 0.25) and CD68 (β: 0.25; 95% CI: 0.10, 0.39) in the third trimester, however was negatively associated with IL-4 (β: -0.21; 95% CI: -0.41, -0.01) and CD206 (β: -0.23; 95% CI: -0.45, -0.02) in the first trimester. Repeated measures analysis showed that TNF-α, IL-6 and CD68 increased by 0.11 (95% CI: 0.01, 0.21), 0.12 (0.15, 95% CI: 0.05, 0.25), 0.22 (95% CI: 0.10, 0.39), respectively, with each 1ln-transformed Tl increase in total samples. Gender-specific analyses revealed that these associations were largely driven by male offspring. In addition, immunohistochemistry revealed that nuclear NF-κB p65 expression increased in placenta tissue. The results of this prospective cohort study provide longitudinal evidence that prenatal Tl exposure induces a placental inflammatory response in the Chinese population., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Both thallium (Tl) and arsenic (As) bear severe toxicity. Brake fern (Pteris vittata L.) is well-known for its hyperaccumulation capacity of As, yet its role on Tl accumulation remains unknown. Herein, brake ferns growing near an As tailing site in Yunnan, Southwestern China are for the first time discovered as a co-hyperaccumulator of both Tl and As. The results showed that the brake ferns extracted both As and Tl efficiently from the soils into the fronds. The studied ferns growing on Tl and As co-polluted soils were found to accumulate extremely high levels of both As (7215-11110 mg/kg) and Tl (6.47-111 mg/kg). Conspicuously high bio-accumulation factor (BCF) was observed for As (7.8) and even higher for Tl (28.4) among these co-hyperaccumulators, wherein the contents of As and Tl in contaminated soils were 1240 ± 12 and 3.91 ± 0.01 mg/kg, respectively. The applied advanced characterization techniques (e.g. transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS)) indicated a preferential uptake of Tl(I) while simultaneous accumulation of As (III) and As(V) from the Tl(I)/Tl(III)-As (III)/As(V) co-existent rhizospheric soils. The findings benefit the phytoremediation practice and pose implications for managing and restoring Tl-As co-contaminated soils in other countries., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We studied thallium (Tl) isotope fractionation in white mustard grown hydroponically at different Tl doses. Thallium isotope signatures in plants indicated preferential incorporation of the light 203 Tl isotope during Tl uptake from the nutrient solution. Negative isotope fractionation was even more pronounced in dependence on how much the available Tl pool decreased. This finding corresponds to the concept of isotope overprinting related to a high contamination level in the growing media (solution or soil). Regarding Tl translocation in plants, we observed a large Tl isotope shift with an enrichment in the heavy 205 Tl isotope in the shoots relative to the roots in treatments with low/moderate solution Tl concentrations (0.01/0.05 mg Tl/L), with the corresponding α 205/203 Tl fractionation factors of ˜1.007 and 1.003, respectively. This finding is probably a consequence of specific (plant) reactions of Tl replacing K in its cycle. The formation of the S-coordinated Tl(I) complexes, potentially affecting both Tl accumulation and Tl isotope fractionation in plants, however, was not proven in our plants, since we did not have indication for that on the basis of X-ray absorption spectroscopy, suggesting that Tl was mainly present as free/hydrated Tl + ion or chemically bound to O-containing functional groups., (Copyright © 2019 Elsevier B.V. All rights reserved.)

It has been suggested that open pit mining and upgrading of bitumen in northern Alberta releases Tl and other potentially toxic elements to the Athabasca River and its watershed. We examined Tl and other trace elements in otoliths of Trout-perch (Percopsis omiscomaycus), a non-migratory fish species, collected along the Athabasca River. Otoliths were analyzed using ICP-QMS, following acid digestion, in the metal-free, ultraclean SWAMP laboratory. Compared to their average abundance in the dissolved (<0.45 μm) fraction of Athabasca River, Tl showed the greatest enrichment in otoliths of any of the trace elements, with enrichments decreasing in the order Tl, Sr, Mn, Zn, Ba, Th, Ni, Rb, Fe, Al, Cr, Ni, Cu, Pb, Co, Li, Y, V, and Mo. Normalizing Tl in the otoliths to the concentrations of lithophile elements such as Li, Rb, Al or Y in the same tissue reveals average enrichments of 177, 22, 19 and 190 times, respectively, relative to the corresponding ratios in the water. None of the element concentrations (Tl, Li, Rb, Al, Y) or ratios were significantly greater downstream of industry compared to upstream. This natural bioaccumulation of Tl most likely reflects the similarity in geochemical and biological properties of Tl + and K + . SUMMARY OF MAIN FINDINGS: Thallium is enriched in fish otoliths, relative to the chemical composition of the river, to the same degree both upstream and downstream of industry., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Thallium (Tl+) flux assays enable imaging of potassium (K+) channel activity in cells and tissues by exploiting the permeability of K+ channels to Tl+ coupled with a fluorescent Tl+ sensitive dye. Common Tl+ sensing dyes utilize fluorescein as the fluorophore though fluorescein exhibits certain undesirable properties in these assays including short excitation wavelengths and pH sensitivity. To overcome these drawbacks, the replacement of fluorescein with rhodols was investigated. A library of 13 rhodol-based Tl+ sensors was synthesized and their properties and performance in Tl+ flux assays evaluated. The dimethyl rhodol Tl+ sensor emerged as the best of the series and performed comparably to fluorescein-based sensors while demonstrating greater pH tolerance in the physiological range and excitation and emission spectra 30 nm red-shifted from fluorescein.

Thallium has been shown to significantly influence various tissues of living organisms; Exposure to Thallium can disturb mitochondrial function, degenerate neurons, and interfere with the function of critical metabolic enzymes and co-enzymes. Glutathione (GSH) an essential biomarker is considered a key factor in harnessing the thallium toxicity. In the present study the interaction of Thallium (Thallium Chloride) and glutathione was investigated spectro-photo-metrically in aqueous media. The renowned Elman's experimental protocol was followed at a wavelength of 412nm for Glutathione quantification in each sample. The pH of each sample was maintained at 7.6 using Phosphate buffer during the entire course of the experiment. A concentration as well as time dependent depletion of glutathione after exposure to various concentration of Thallium metal was observed, revealing chemical interaction between the metal and glutathione. The exact mechanism of interaction of Thallium and glutathione is still to be investigated. However, this piece of research suggests that a decrease in the concentration of Glutathione may be due to Thallium-GSH abduct or oxidize glutathione (GSSG) formation. This study was performed in-vitro as a model of in vivo.

TRPM5 functions as a calcium-activated monovalent cation-selective ion channel and is expressed in a variety of cell types. Dysfunction of this type of channel has been recently implied in cardiac arrhythmias, diabetes, and other pathologies. Therefore, a growing interest has emerged to develop the pharmacology of these ion channels. We optimized a screening assay based on the thallium flux through the TRPM5 channel and a fluorescent thallium dye as a probe for channel activity. We show that this assay is capable of identifying molecules that inhibit or potentiate calcium-activated monovalent cation-selective ion channels.

Thallium (Tl) is a well-recognized hazardous toxic heavy metal that has been reported to have embryotoxicity and fetotoxicity. However, little is known about its association with preterm birth (PTB) in humans. We aimed to evaluate the predictors of Tl exposure and assessed its relation with PTB. The study population included 7173 mother-infant pairs from a birth cohort in Wuhan, China. Predictors of Tl concentrations were explored using linear regression analyses, and associations of Tl exposure with risk of PTB or gestational age at birth were estimated using logistic regression or generalized linear models. The geometric mean and median values of urinary Tl concentrations were 0.28 μg/L (0.55 μg/g creatinine) and 0.29 μg/L (0.53 μg/g creatinine). We found that maternal urinary Tl concentrations varied by gestational weight gain, educational attainment, multivitamin and iron supplementations. Women with Tl concentrations higher than 0.80 μg/g creatinine were at higher risk of giving birth prematurely versus those with Tl concentrations lower than 0.36 μg/g creatinine [adjusted odds ratio (95% confidence interval (CI)): 1.55 (1.05, 2.27)], and the association was more pronounced in PTB with premature rupture of membranes (PROM) rather than in PTB without PROM. About 3-fold increase in creatinine-corrected Tl concentrations were associated with 0.99-day decrease in gestational length (95% CI: -1.36, -0.63). This is the first report on the associations between maternal Tl exposure and the risk of PTB., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

To investigate the bioavailability of thallium (Tl) in soil and rice in a Tl-contaminated area in Guangdong, China, the topsoil and rice samples were collected from 24 sampling sites and analyzed. Moreover, a modified sequential extraction procedure was applied to determine the different Tl fractions in the soil. The mean pH value of the soil samples was 4.50. The total Tl concentration in the paddy soil was about 4-8 times higher than the Canadian guideline value (1mgkg -1 ) for agricultural land uses. The mean ecological risk index of Tl was determined to be 483, indicating that potential hazard of the paddy soil was serious. The mean content of Tl in rice was 1.42mgkg -1 , which exceeded the German maximum permissible level (0.5mgkg -1 ) of Tl in foods and feedstuffs by a factor of nearly 3. The hazard quotient value via rice intake was 57.6, indicating a high potential health risk to the local residents. The distribution of various Tl fractions followed the order of easily reducible fraction (40.3%) > acid exchangeable fraction (30.5%) > residual fraction (23.8%) > oxidizable fraction (5.4%). Correlation analyses showed that the easily reducible fraction correlates positively with the soil Fe and Mn contents, whereas the acid exchangeable fraction is significantly correlated with the S content. The soil pH was negatively correlated with the Tl content in both soil and rice. The Tl content in rice was more strongly correlated with the exchangeable fraction than the total Tl content in the soil. Overall, the bioavailability of Tl in more acidic soil is higher, and is strongly dependent on the speciation of Tl, especially the content of acid exchangeable fraction., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Thallium (Tl + ) is a ubiquitous natural trace metal considered as the most toxic among heavy metals. The ionic ratio of Tl + is similar to that of potassium (K + ), therefore accounting for the replacement of the latter during enzymatic reactions. The principal organelle damaged after Tl + exposure is mitochondria. Studies on the mechanisms of Tl + include intrinsic pathways altered and changes in antiapoptotic and proapoptotic proteins, cytochrome c, and caspases. Oxidative damage pathways increase after Tl + exposure to produce reactive oxygen species (ROS), changes in physical properties of the cell membrane caused by lipid peroxidation, and concomitant activation of antioxidant mechanisms. These processes are likely to account for the neurotoxic effects of the metal. In humans, Tl + is absorbed through the skin and mucous membranes and then is widely distributed throughout the body to be accumulated in bones, renal medulla, liver, and the Central Nervous System. Given the growing relevance of Tl + intoxication, in recent years there is a notorious increase in the number of reports attending Tl + pollution in different countries. In this sense, the neurological symptoms produced by Tl + and its neurotoxic effects are gaining attention as they represent a serious health problem all over the world. Through this review, we present an update to general information about Tl + toxicity, making emphasis on some recent data about Tl + neurotoxicity, as a field requiring attention at the clinical and preclinical levels.

Gümüsköy Ag (As, Pb, and Tl) deposits are one of the largest silver deposits in the country and located about 25 km west of Kütahya, Turkey. This study investigated the accumulation and transport of thallium into 11 wild plants in soil of the mining area. Plant samples and their associated soils were collected from the field and Tl contents were measured with inductively coupled plasma mass spectroscopy (ICP-MS). The mean concentrations in the soil, roots, and shoots of the studied plants were, respectively, 170, 318, and 315 mg kg(-1) for Tl. The plants analyzed and collected from the studied area were separated into different groups based on enrichment coefficients of roots and shoots (ECR and ECS). The results showed that because of their higher ECR and ECS, the following could be good bioaccumulators: CY, IS, SL, and VR for Tl. Therefore, these plants can be useful for remediation or phytoremediation of soils polluted by Tl.

Background: Anesthetic preconditioning (APC) is a clinically important phenomenon in which volatile anesthetics (VAs) protect tissues such as heart against ischemic injury. The mechanism of APC is thought to involve K channels encoded by the Slo gene family, and the authors showed previously that slo-2 is required for APC in Caenorhabditis elegans. Thus, the authors hypothesized that a slo-2 ortholog may mediate APC-induced cardioprotection in mammals., Methods: A perfused heart model of ischemia-reperfusion injury, a fluorescent assay for K flux, and mice lacking Slo2.1 (Slick), Slo2.2 (Slack), or both (double knockouts, Slo2.x dKO) were used to test whether these channels are required for APC-induced cardioprotection and for cardiomyocyte or mitochondrial K transport., Results: In wild-type (WT) hearts, APC improved post-ischemia-reperfusion functional recovery (APC = 39.5 ± 3.7% of preischemic rate × pressure product vs. 20.3 ± 2.3% in controls, means ± SEM, P = 0.00051, unpaired two-tailed t test, n = 8) and lowered infarct size (APC = 29.0 ± 4.8% of LV area vs. 51.4 ± 4.5% in controls, P = 0.0043, n = 8). Protection by APC was absent in hearts from Slo2.1 mice (% recovery APC = 14.6 ± 2.6% vs. 16.5 ± 2.1% in controls, P = 0.569, n = 8 to 9, infarct APC = 52.2 ± 5.4% vs. 53.5 ± 4.7% in controls, P = 0.865, n = 8 to 9). APC protection was also absent in Slo2.x dKO hearts (% recovery APC = 11.0 ± 1.7% vs. 11.9 ± 2.2% in controls, P = 0.725, n = 8, infarct APC = 51.6 ± 4.4% vs. 50.5 ± 3.9% in controls, P = 0.855, n = 8). Meanwhile, Slo2.2 hearts responded similar to WT (% recovery APC = 41.9 ± 4.0% vs. 18.0 ± 2.5% in controls, P = 0.00016, n = 8, infarct APC = 25.2 ± 1.3% vs. 50.8 ± 3.3% in controls, P < 0.000005, n = 8). Furthermore, VA-stimulated K transport seen in cardiomyocytes or mitochondria from WT or Slo2.2 mice was absent in Slo2.1 or Slo2.x dKO., Conclusion: Slick (Slo2.1) is required for both VA-stimulated K flux and for the APC-induced cardioprotection.

Aim: To establish an improved, high-throughput screening techniques for identifying novel KCNQ2 channel activators., Methods: KCNQ2 channels were stably expressed in CHO cells (KCNQ2 cells). Thallium flux assay was used for primary screening, and 384-well automated patch-clamp IonWorks Barracuda was used for hit validation. Two validated activators were characterized using a conventional patch-clamp recording technique., Results: From a collection of 80 000 compounds, the primary screening revealed a total of 565 compounds that potentiated the fluorescence signals in thallium flux assay by more than 150%. When the 565 hits were examined in IonWorks Barracuda, 38 compounds significantly enhanced the outward currents recorded in KCNQ2 cells, and were confirmed as KCNQ2 activators. In the conventional patch-clamp recordings, two validated activators ZG1732 and ZG2083 enhanced KCNQ2 currents with EC50 values of 1.04±0.18 μmol/L and 1.37±0.06 μmol/L, respectively., Conclusion: The combination of thallium flux assay and IonWorks Barracuda assay is an efficient high-throughput screening (HTS) route for discovering KCNQ2 activators.

Thallium (Tl) is a highly volatile and toxic heavy metal regarded to cause pollution even at very low concentrations of several parts per million. Despite the extremely high risk of Tl in the environment, limited information on removal/recovery exists. The study focussed on the use of green algae to determine the sorption potential and recovery of Tl. From the study, removal efficiency was achieved at 100% for lower concentrations of ≥150 mg/L of Tl. At higher concentrations in a range of 250-500 mg/L, the performance of algae was still higher with sorption capacity (qmax) between 830 and 1000 mg/g. Generally, Chlorella vulgaris was the best adsorbent with a high qmax and lower affinity of 1000 mg/g and 1.11 L/g, respectively. When compared to other studies on Tl adsorption, the tested algae showed a better qmax than most adsorbents. The kinetic studies showed better correlation co-efficient of ≤0.99 for Pseudo-second order model than the first order model. Recovery was achieved highest for C. vulgaris using nitric acid at 93.3%. The strongest functional groups responsible for Tl binding on the algal cell wall were carboxyl and phenols. Green algae from freshwater bodies showed significant potential for Tl removal/recovery from industrial wastewater., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Little is known about the biosorption and bioaccumulation capacity of thallium (Tl) by microorganisms that occur in Tl-polluted soil. The present study focused on characterizing the biosorption and bioaccumulation of Tl by Tl-tolerant fungi isolated from Tl-polluted soils. Preliminary data showed a positive correlation between the biomass and the biosorbed Tl content. The Tl-tolerant strains were capable of bioaccumulating Tl, up to 7189 mg kg(-1) dry weight. The subcellular distribution of Tl showed obvious compartmentalization: cytoplasm ≫ cell wall > organelle. The majority of Tl (up to 79%) was found in the cytoplasm, suggesting that intracellular compartmentalization appeared to be responsible for detoxification. These findings further suggest the applicability of the fungal isolates for cleanup of Tl in Tl-polluted water and soil.

Thallium (Tl) is a highly toxic heavy metal situated between mercury and lead in the periodic table. While its neighbors have been thoroughly studied for DNA-based sensing, little is known about thallium detection. In this work, in vitro selection of RNA-cleaving DNAzymes is carried out using Tl(3+) as the target metal cofactor. Both normal DNA and phosphorothioate (PS)-modified DNA are tested for this purpose. While no Tl(3+)-dependent DNAzymes are obtained, a DNA oligonucleotide containing a single PS-modified RNA nucleotide is found to cleave by ∼7% by Tl(3+) at the RNA position. The remaining 93% are desulfurized. By hybridization of this PS-modified oligonucleotide with the Tm7 DNAzyme, the cleavage yield increases to ∼40% in the presence of Tl(3+) and Er(3+). Tm7 is an Er(3+)-dependent RNA-cleaving DNAzyme. It cleaves only the normal substrate but is completely inactive using the PS-modified substrate. Tl(3+) desulfurizes the PS substrate to the normal substrate to be cleaved by Tm7 and Er(3+). This system is engineered into a catalytic beacon for Tl(3+) with a detection limit of 1.5 nM, which is below its maximal contamination limit defined by the U.S. Environmental Protection Agency (10 nM).

Thallium (Tl) pollution in agricultural areas can pose hidden danger to humans, as food consumption is the key exposure pathway of Tl. Owing to the extreme toxicity of Tl, removal of Tl from soil becomes necessary to minimize the Tl-related health effects. Phytoremediation is a cost-effective method to remove heavy metals from soil, but not all plants are appropriate for this purpose. Here, the ability of Solanum nigrum L., commonly known as black nightshade, to remediate Tl-contaminated soil was evaluated. The accumulation of Tl in different organs of S. nigrum was measured under both field and greenhouse conditions. Additionally, the growth and maximal quantum efficiency of photosystem II (Fv/Fm) under different Tl concentrations (1, 5, 10, 15, and 20 mg kg(-1)) were examined after 4-month pot culture. Under both field and greenhouse conditions, Tl accumulated in S. nigrum was positively correlated with Tl concentration in the soil. Thallium mostly accumulated in the root, and bioconcentration factor was greater than 1, indicating the good capability of S. nigrum to extract Tl. Nonetheless, the growth and Fv/Fm of S. nigrum were reduced at high Tl concentration (>10 mg kg(-1)). Given the good tolerance, fast growth, high accumulation, and global distribution, we propose that S. nigrum is a competent candidate to remediate moderately Tl-contaminated soil (<10 mg kg(-1)) without causing far-reaching ecological consequences.

Ion channels are critical for life, and they are targets of numerous drugs. The sequencing of the human genome has revealed the existence of hundreds of different ion channel subunits capable of forming thousands of ion channels. In the face of this diversity, we only have a few selective small-molecule tools to aid in our understanding of the role specific ion channels in physiology which may in turn help illuminate their therapeutic potential. Although the advent of automated electrophysiology has increased the rate at which we can screen for and characterize ion channel modulators, the technique's high per-measurement cost and moderate throughput compared to other high-throughput screening approaches limit its utility for large-scale high-throughput screening. Therefore, lower cost, more rapid techniques are needed. While ion channel types capable of fluxing calcium are well-served by low cost, very high-throughput fluorescence-based assays, other channel types such as sodium channels remain underserved by present functional assay techniques. In order to address this shortcoming, we have developed a thallium flux-based assay for sodium channels using the NaV1.7 channel as a model target. We show that the assay is able to rapidly and cost-effectively identify NaV1.7 inhibitors thus providing a new method useful for the discovery and profiling of sodium channel modulators.

Glucose 6-phosphate dehydrogenase (d-glucose 6-phosphate: NADP(+) oxidoreductase, EC 1.1.1.49; G6PD) is a key enzyme that is localized in all mammal tissues, especially in cytoplasmic sections and that catalyzes the first step of pentose phosphate metabolic pathway. In this study, G6PD enzyme was purified 1444-fold with a yield of 77% from rainbow trout liver using 2',5'-ADP-sepharose-4B affinity chromatography. Moreover, a purity check of the enzyme was performed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Some characteristic features like optimal pH, stable pH, optimal temperature and optimal ionic strength were determined for the purified enzyme. In addition to this, in vitro effects of ions like silver nitrate (Ag(+)), thallium sulphate (TI(+)), cobalt (II) nitrate (Co(2+)) and arsenic (V) oxide (As(5+)) on enzyme activity were researched. Half-maximal inhibitory concentration (IC50) values of Ag(+), Co(2+) and As(5+) metal ions, which showed an inhibitory effect, were found to be 0.0044, 0.084 and 4.058 mM, respectively; and their inhibition constants (K i) were found to be 0.0052 ± 0.00042, 0.087 ± 0.015700 and 4.833 ± 1.753207 mM, respectively. Tl(+) not exhibited inhibitory effect on the enzyme activity., (© The Author(s) 2013.)

Thallium (Tl) is very toxic to mammals but little is known about its accumulation by plants, and it would be useful if prediction of Tl accumulation could be done using potassium (K) accumulation models. The objectives of this study were to compare the uptake kinetics of Tl(+) and K(+), and to determine how readily K(+) can inhibit Tl(+) uptake. Durum wheat (Triticum turgidum L.) and spring canola (Brassica napus L.) were grown hydroponically and exposed to 0-75 μM Tl or 0-250 μM K for up to 150 min (kinetics experiment), or to 0.1 or 10 μM Tl with Tl to K ratios of 1:1 to 1:10,000 for up to 300 min (competition experiment). The rate of uptake of Tl(+) by canola was about three to five times faster than by wheat, while the rate of Tl(+) uptake in wheat was the same as the rate of K(+) uptake by either species. Uptake of Tl(+) was more readily suppressed by K(+) in wheat than in canola. When exposed to 0.1 uM Tl for 300 min with 100 or 1,000 uM K(+), Tl(+) uptake by wheat was reduced by 20 % and 50 %, respectively, while Tl(+) uptake by canola was not reduced. Our results suggest that predicting Tl accumulation using a K accumulation model with a correction factor may be possible for canola, but would be much more difficult for wheat, since uptake of Tl(+) is very sensitive to levels of K(.)

The accumulation of thallium (Tl) in brassicaceous crops is widely known, but both the uptake extents of Tl by the individual cultivars of green cabbage and the distribution of Tl in the tissues of green cabbage are not well understood. Five commonly available cultivars of green cabbage grown in the Tl-spiked pot-culture trials were studied for the uptake extent and subcellular distribution of Tl. The results showed that all the trial cultivars mainly concentrated Tl in the leaves (101∼192 mg/kg, DW) rather than in the roots or stems, with no significant differences among cultivars (p = 0.455). Tl accumulation in the leaves revealed obvious subcellular fractionation: cell cytosol and vacuole >> cell wall > cell organelles. The majority (∼ 88%) of leaf-Tl was found to be in the fraction of cytosol and vacuole, which also served as the major storage site for other major elements such as Ca and Mg. This specific subcellular fractionation of Tl appeared to enable green cabbage to avoid Tl damage to its vital organelles and to help green cabbage tolerate and detoxify Tl. This study demonstrated that all the five green cabbage cultivars show a good application potential in the phytoremediation of Tl-contaminated soils.

A breeding population of the common toad Bufo bufo living in the vicinity of a Zn-Pb smelting works in Bukowno, Poland was studied for the presence of thallium. Tl concentration was measured in the bottom sediments of the spawning pond, in the laid eggs, in juveniles after metamorphosis, and in the selected tissues of the adult individuals. A very high concentration of Tl was detected in the spawn (13.97 ± 8.90 mg/kg d.w.). In 50% of the spawn samples, levels exceeded 20 mgTl/kg d.w. The issue of maternal transfer of thallium from females to oocytes is discussed. Due to a significant accumulation of thallium, spawn analysis can be used as a sensitive indicator of the presence of this element in the environment and may replace more invasive methods that involve the killing of adult animals. In those regions that are abundant in Zn-Pb ores, the spawn of amphibians may be a very important source of thallium contamination for predators. From among all tissues of the Bukowno adult toads, the livers have shown the highest accumulation of thallium (mean 3.98 mg/kg d.w. and maximum value--18.63). For as many as 96.5% of livers, concentrations exceeded 1.0 mgTl/kg d.w. which is treated as indicative of poisoning.

The purpose of this study is to assess the long-term stability of Prussian blue (PB) drug product (DP) and active pharmaceutical ingredient (API) under laboratory storage conditions by monitoring the loss in water content and the corresponding change of the in vitro thallium binding capacity that represents product performance. The bound water content and the in vitro thallium binding capacity of PB DPs and APIs were measured in 2003 and 2013, respectively. Water content, a critical quality attribute that directly correlates to the thallium (Tl) binding capacity was measured by thermal gravimetric analysis (TGA). The thallium binding study was conducted by testing PB in buffered solutions over the human gastrointestinal pH range with thallium concentrations ranging from 600 to 1,500 ppm. Samples were incubated at physiological temperature of 37°C in a shaking water bath to mimic gastric flux and intestinal transport. The binding equilibrium was reached at 24h. Following incubation, each sample was filtered and the free thallium was analyzed using a validated inductively coupled plasma spectroscopic method (ICP). The Langmuir isotherm was plotted to calculate maximum binding capacity (MBC). Compared with 2003, the water content of DP-1 decreased by about 14.1% (from 15.6 to 13.4 mol), and the MBC of DP-1 decreased by about 12.5% (from 714 to 625 mg/g) at pH 7.5. When low concentration of thallium (600 ppm) was used at pH 7.5, the Tl binding remained comparable for both API-1 (286 vs 276 mg/g) and DP-1 (286 vs 268 mg/g). Similarly, the Tl binding remained unchanged for both API-1 (237 vs 255 mg/g) and DP-1 (234 vs 236 mg/g) at pH 5.0. However, at pH 1.0 the binding was reduced 32.3% and 25.9% for API-1 and DP-1, respectively. Since the majority of binding takes place in the upper GI tract where pH around 5 can be expected, and therefore, the Tl binding capacity of PB should be comparable for new and aged samples. The findings that Tl binding changes with the water loss of PB and pH conditions are consistent with our previously published data. The study also represents the first quantitative assessment of the long-term stability of PB. Over last 10 years, PB DPs and APIs have lost about 20% water under ambient laboratory storage conditions which are consistent with a controlled warehouse environment. While the maximum binding capacity of PB to thallium was decreased after about 10 years of long-term storage, it is still very effective, suggesting that the shelf life of PB should be much longer than the manufacturer ascribed expiration date of 2008 under proper storage conditions., (Published by Elsevier B.V.)

Metabotropic glutamate receptor 7 (mGlu7) is a member of the group III mGlu receptors (mGlus), encompassed by mGlu4, mGlu6, mGlu7, and mGlu8. mGlu7 is highly expressed in the presynaptic active zones of both excitatory and inhibitory synapses, and activation of the receptor regulates the release of both glutamate and GABA. mGlu7 is thought to be a relevant therapeutic target for a number of neurological and psychiatric disorders, and polymorphisms in the GRM7 gene have been linked to autism, depression, ADHD, and schizophrenia. Here we report two new pan-group III mGlu positive allosteric modulators, VU0155094 and VU0422288, which show differential activity at the various group III mGlus. Additionally, both compounds show probe dependence when assessed in the presence of distinct orthosteric agonists. By pairing studies of these nonselective compounds with a synapse in the hippocampus that expresses only mGlu7, we have validated activity of these compounds in a native tissue setting. These studies provide proof-of-concept evidence that mGlu7 activity can be modulated by positive allosteric modulation, paving the way for future therapeutics development.

We showed earlier that diminution of 2,4-dinitrophenol (DNP)-stimulated respiration and increase of both mitochondrial swelling and electrochemical potential (ΔΨmito) dissipation in medium containing TlNO3 and KNO3 were caused by opening of Tl(+)-induced mitochondrial permeability transition pore (MPTP) in the inner membrane of Ca(2+)-loaded rat liver mitochondria. The MPTP opening was studied in the presence of bivalent metal ions (Sr(2+), Ba(2+), Mn(2+), Co(2+) and Ni(2+)), trivalent metal ions (Y(3+) and La(3+)), and ruthenium red. We found that these metal ions (except Ba(2+) and Co(2+)) as well as ruthenium red inhibited to the MPTP opening that manifested in preventing both diminution of the DNP-stimulated respiration and increase of the swelling and of the ΔΨmito dissipation in medium containing TlNO3, KNO3, and Ca(2+). Inhibition of the MPTP opening by Sr(2+) and Mn(2+) is suggested because of their interaction with high affinity Ca(2+) sites, facing the matrix side and participating in the MPTP opening. The inhibitory effects of metal ions (Y(3+), La(3+), and Ni(2+)), and ruthenium red are accordingly discussed in regard to competitive and noncompetitive inhibition of the mitochondrial Ca(2+)-uniporter. High concentrations (50μM) of Y(3+) and La(3+) favored of MPTP opening in the inner membrane of rat liver mitochondria in Ca(2+) free medium containing TlNO3. The latter MPTP opening was markedly eliminated by MPTP inhibitors (cyclosporine A and ADP)., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The objective of the present work was to study the phytoremediation capacity of Callitriche cophocarpa concerning water contaminated with thallium (Tl), cadmium (Cd), zinc (Zn), and lead (Pb) derived from the natural environment. We found that after a 10-day incubation period, shoots of C. cophocarpa effectively biofiltrated the water so that it met (for Cd, Zn, and Pb) appropriate quality standards. The order of accumulation of the investigated elements by shoots (mg kg(-1) dry weight) were as follows: Zn (1120) < Tl (251) < Cd (71) < Pb (35). The order of bioconcentration factors were as follows: Cd (1177) < Tl (1043) < Zn (718) < Pb (597). According to Microtox bioassay, C. cophocarpa significantly eradicated polluted water toxicity. During the experiment, the physiological status of plants was monitored by taking measurements of photosystem II activity (maximum efficiency of PSII, photochemical fluorescence quenching, nonphotochemical fluorescence quenching, and quantum efficiency of PSII), photosynthetic pigment contents, and shoot morphology. Plants exposed to metallic pollution did not exhibit significant changes in their physiological status compared with the control. This work is potentially applicable to the future use of C. cophocarpa in the phytoremediation of polluted, natural watercourses.

Allchar mine is an abandoned arsenic-antimony-thallium deposit located on the northwestern part of Kozuf Mt., Republic of Macedonia. Allchar is a unique deposit within the world, due to the variety of its mineral composition especially and in the high content of thallium. The aim of this work was to assess the level of contamination at this post-mining area as well as to determine the intensity of accumulation of various elements (Ag, Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Ga, K, Li, Mg, Mn, Mo, Na, Ni, P, Pb, Rb, S, Sb, Sr, Tl, V, and Zn) with focus on As, Sb and Tl, in two endemic Viola species from this locality (Viola allcharensis G. Beck, Viola arsenica G. Beck) and one Balkan endemic species (Viola macedonica Boiss. & Heldr.). Samples of different plant parts and soil were digested and then analysed by ICP-AES. It was found that the accumulation of As, Sb, and Tl in these endemic species is significantly high. In this study a systematic investigation of the As-Sb-Tl contamination of soils and their bioavailability was carried out using the extraction procedure in order to explore the mobility and potential bioavailability of the As, Sb, and Tl.

Thallium is a highly toxic metal whose biogeochemical behaviour in the marine environment is poorly understood. We measured Tl in sediments, macroalgae (Fucus vesiculosus and Fucus ceranoides) and deposit-feeding invertebrates (Hediste diversicolor, Arenicola marina and Scrobicularia plana) from two estuaries of south west England (Plym and Fal) draining mineralised catchments. In the Plym, and for a given sample type, concentrations of Tl were rather invariant between sample locations and averaged about 500 μg kg(-1) for sediment, 30 μg kg(-1) for macroalgae and 10 μg kg(-1) for the invertebrates. In the Fal, respective concentrations were of a similar order of magnitude but exhibited greater variation between sample locations. Normalisation of Tl concentrations to K, the biogeochemical analogue of Tl(+), revealed bioenrichment of about 20 for all organisms in the Plym and bioenrichment ranging from about 3 (H. diversicolor) to 170 (F. ceranoides) in the Fal. Despite the low bioaccumulation of Tl relative to other metals measured concurrently, it is recommended that Tl be more closely monitored and better studied in the estuarine environment., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Specific members of the inward rectifier potassium (Kir) channel family are postulated drug targets for a variety of disorders, including hypertension, atrial fibrillation, and pain. For the most part, however, progress toward understanding their therapeutic potential or even basic physiological functions has been slowed by the lack of good pharmacological tools. Indeed, the molecular pharmacology of the inward rectifier family has lagged far behind that of the S4 superfamily of voltage-gated potassium (Kv) channels, for which a number of nanomolar-affinity and highly selective peptide toxin modulators have been discovered. The bee venom toxin tertiapin and its derivatives are potent inhibitors of Kir1.1 and Kir3 channels, but peptides are of limited use therapeutically as well as experimentally due to their antigenic properties and poor bioavailability, metabolic stability and tissue penetrance. The development of potent and selective small-molecule probes with improved pharmacological properties will be a key to fully understanding the physiology and therapeutic potential of Kir channels. The Molecular Libraries Probes Production Center Network (MLPCN) supported by the National Institutes of Health (NIH) Common Fund has created opportunities for academic scientists to initiate probe discovery campaigns for molecular targets and signaling pathways in need of better pharmacology. The MLPCN provides researchers access to industry-scale screening centers and medicinal chemistry and informatics support to develop small-molecule probes to elucidate the function of genes and gene networks. The critical step in gaining entry to the MLPCN is the development of a robust target- or pathway-specific assay that is amenable for high-throughput screening (HTS). Here, we describe how to develop a fluorescence-based thallium (Tl(+)) flux assay of Kir channel function for high-throughput compound screening. The assay is based on the permeability of the K(+) channel pore to the K(+) congener Tl(+). A commercially available fluorescent Tl(+) reporter dye is used to detect transmembrane flux of Tl(+) through the pore. There are at least three commercially available dyes that are suitable for Tl(+) flux assays: BTC, FluoZin-2, and FluxOR. This protocol describes assay development using FluoZin-2. Although originally developed and marketed as a zinc indicator, FluoZin-2 exhibits a robust and dose-dependent increase in fluorescence emission upon Tl(+) binding. We began working with FluoZin-2 before FluxOR was available and have continued to do so. However, the steps in assay development are essentially identical for all three dyes, and users should determine which dye is most appropriate for their specific needs. We also discuss the assay's performance benchmarks that must be reached to be considered for entry to the MLPCN. Since Tl(+) readily permeates most K(+) channels, the assay should be adaptable to most K(+) channel targets.