Effects of Acute and Subchronic Waterborne Thallium Exposure on Ionoregulatory Enzyme Activity and Oxidative Stress in Rainbow Trout (Oncorhynchus mykiss).

The mechanisms of acute (96-hour) and subchronic (28-day) toxicity of the waterborne trace metal thallium (Tl) to rainbow trout (Oncorhynchus mykiss) were investigated. Specifically, effects on branchial and renal ionoregulatory enzymes (sodium/potassium adenosine triphosphatase [ATPase; NKA] and proton ATPase) and hepatic oxidative stress endpoints (protein carbonylation, glutathione content, and activities of catalase and glutathione peroxidase) were examined. Fish (19-55 g) were acutely exposed to 0 (control), 0.9 (regulatory limit), 2004 (half the acute median lethal concentration), or 4200 (acute median lethal concentration) µg Tl L-1 or subchronically exposed to 0, 0.9, or 141 (an elevated environmental concentration) µg Tl L-1 . The only effect following acute exposure was a stimulation of renal H+ -ATPase activity at the highest Tl exposure concentration. Similarly, the only significant effect of subchronic Tl exposure was an inhibition of branchial NKA activity at 141 µg Tl L-1 , an effect that may reflect the interaction of Tl with potassium ion handling. Despite significant literature evidence for effects of Tl on oxidative stress, there were no effects of Tl on any such endpoint in rainbow trout, regardless of exposure duration or exposure concentration. Elevated basal levels of antioxidant defenses may explain this finding. These data suggest that ionoregulatory perturbance is a more likely mechanism of Tl toxicity than oxidative stress in rainbow trout but is an endpoint of relevance only at elevated environmental Tl concentrations. Environ Toxicol Chem 2024;43:87-96. © 2023 SETAC.

Mechanistic examination of thallium and potassium interactions in Daphnia magna.

The trace element thallium (Tl) exerts its toxic effects, at least in part, through its mimicry of potassium (K+) and subsequent impairment of K+ homeostasis. However, the specific nature of this effect remains poorly understood, especially in aquatic biota that are threatened by elevated concentrations of Tl associated with mining and refining effluents. In this study experiments were conducted to mechanistically examine the relationship between Tl and K+ in terms of uptake and toxicity in the regulatory model species Daphnia magna. In one set of experiments the effects of K+, the K+ analog rubidium (Rb+), and generalized K+ channel blocker cesium (Cs+) on Tl-induced acute toxicity were examined. The presence of increasing concentrations of K+ and Rb+ in exposure water reduced waterborne Tl toxicity, indicating that the actions of Tl were mediated at least in part through interactions with K+. However, in the presence of elevated water Cs+, the toxicity of Tl paradoxically increased. Pharmaceuticals with putative blocking actions on K+ channels failed to alter whole-body K+ of control organisms, but in the case of clozapine and chlorpropamide, whole-body K+ status was significantly elevated relative to exposures with Tl alone, which tended to reduce this metric. These data identify inwardly rectifying and voltage gated K+ channels as potential loci of Tl toxicity. Experiments using rubidium (Rb+) as a tracer of K+, showed that waterborne Tl affects the uptake of K+, but the magnitude of inhibition by Tl was not sufficient to explain the effect on whole-body K+. While these data indicate interactions between Tl and K occur at K+ transporters in D magna, they also indicate that environmental levels of K+ are likely to ameliorate toxicity in most natural waters.

Accumulation of Thallium in Rainbow Trout (Oncorhynchus mykiss) Following Acute and Subchronic Waterborne Exposure.

The accumulation and tissue distribution of toxicants in aquatic biota can be determinative of their toxic impact to both exposed organisms and their potential human consumers. In the present study, accumulation of the trace metal thallium (Tl) in gill, muscle, plasma, and otoliths of rainbow trout (Oncorhynchus mykiss) following acute (96-h) and subchronic (28-day) waterborne exposures was investigated. Owing to known interactions between Tl and potassium ions (K+ ), plasma and muscle K+ concentrations were also determined. Branchial Tl accumulated in a dose-dependent manner in both acute and subchronic exposures, while plasma Tl was rapidly mobilized to tissues and accumulated only at exposure concentrations of 141 µg L-1 or higher. For muscle tissue, Tl concentrations at 28 days were markedly lower than those at 96 h at comparable exposure concentrations (0.9 µg L-1 ), indicating the presence of mechanisms that act to reduce Tl accumulation over time. However, after acute exposure, muscle Tl reached concentrations that, if consumed, would exceed acceptable daily intake values for this element, indicating some risk to human health from the consumption of fish from waters heavily contaminated with Tl. Otoliths showed Tl concentrations that reflected exposure concentration and length, confirming their capacity to provide insight into fish exposure history. No changes in tissue K+ concentrations were observed, suggesting that accumulation of Tl in rainbow trout plasma and muscle does not occur at the expense of K+ homeostasis. In addition to highlighting the capacity of rainbow trout to accumulate Tl to levels that exceed recommended dietary doses to human consumers, the present study provides the first data of tissue-specific Tl accumulation in an important regulatory species. Environ Toxicol Chem 2023;42:1553-1563. © 2023 SETAC.

Parallel computing for mobilities in periodic geometries.

We examine methods for calculating the effective mobilities of molecules driven through periodic geometries in the context of particle-based simulation. The standard formulation of the mobility, based on the long-time limit of the mean drift velocity, is compared to a formulation based on the mean first-passage time of molecules crossing a single period of the system geometry. The equivalence of the two definitions is derived under weaker assumptions than similar conclusions obtained previously, requiring only that the state of the system at subsequent period crossings satisfy the Markov property. Approximate theoretical analyses of the computational costs of estimating these two mobility formulations via particle simulations suggest that the definition based on first-passage times may be substantially better suited to exploiting parallel computation hardware. This claim is investigated numerically on an example system modeling the passage of nanoparticles through the slit-well device. In this case, the traditional mobility formulation is found to perform best when the Péclet number is small, whereas the mean first-passage time formulation is found to converge much more quickly when the Péclet number is moderate or large. The results suggest that, given relatively modest access to modern GPU hardware, this alternative mobility formulation may be an order of magnitude faster than the standard technique for computing effective mobilities of biomolecules through periodic geometries.

Studying first passage problems using neural networks: A case study in the slit-well microfluidic device.

This study presents deep neural network solutions to a time-integrated Smoluchowski equation modeling the mean first passage time of nanoparticles traversing the slit-well microfluidic device. This physical scenario is representative of a broader class of parametrized first passage problems in which key output metrics are dictated by a complicated interplay of problem parameters and system geometry. Specifically, whereas these types of problems are commonly studied using particle simulations of stochastic differential equation models, here the corresponding partial differential equation model is solved using a method based on deep neural networks. The results illustrate that the neural network method is synergistic with the time-integrated Smoluchowski model: together, these are used to construct continuous mappings from key physical inputs (applied voltage and particle diameter) to key output metrics (mean first passage time and effective mobility). In particular, this capability is a unique advantage of the time-integrated Smoluchowski model over the corresponding stochastic differential equation models. Furthermore, the neural network method is demonstrated to easily and reliably handle geometry-modifying parameters, which is generally difficult to accomplish using other methods.

Effect of thallium on phototactic behaviour in Daphnia magna.

Thallium (Tl) is a trace metal enriched in wastewaters associated with mining and smelting of base metals. The toxicity of Tl to aquatic biota is poorly understood, particularly with respect to its sublethal effects. In this study, phototactic behavioural responses of naïve (i.e. no previous exposure to Tl) Daphnia magna, a key regulatory freshwater crustacean species, were examined in waters containing Tl. Fed and fasted neonate daphnids (< 24 h old) and fed adults (10-15 days old) showed no significant response at any tested water Tl concentration. However, in fasted adults, an increase in the positive phototactic response (measured as a greater number of daphnids closer to the light source after a 5-min exposure) was seen at Tl concentrations of 917 and 2099 µg L-1, values representative of extreme environmental Tl concentrations. The presence of Tl also decreased the swimming speed of adult Daphnia towards a light source. In the presence of cimetidine, a histamine receptor blocker, the increase in positive phototaxis induced by Tl disappeared, suggesting that Tl acts to perturb the phototaxis response through sensory inhibition. Conversely, although there was a trend towards enhanced activity, Tl had no significant effect on acetylcholinesterase, a marker of locomotor capacity.

Chronic toxicity of waterborne thallium to Daphnia magna.

There is limited information regarding the toxicity of the trace element thallium (Tl) to aquatic biota, most of which assesses acute toxicity and bioaccumulation. The relative lack of chronic Tl toxicity data compromises the establishment of water quality criteria for this trace metal. In the presented work, chronic toxicity endpoints (final body weight (a proxy measure of growth), survival, and reproduction) and Tl body burden were measured in the freshwater crustacean Daphnia magna during a 21-day exposure to dissolved Tl. Thallium caused complete mortality in daphnids between exposure concentrations of 424 and 702 µg L-1. In contrast with previously published work examining acute Tl toxicity, exposure to Tl for 21 days was not associated with changes in whole-body potassium concentration. This was despite a 710-fold increase in Tl body burden in animals exposed to 424 µg L-1 relative to the control. Median effect concentrations (EC50's) for growth and reproduction (total neonates produced), were 1.6 (95% confidence interval: 1.0-3.1) and 11.1 (95% confidence interval: 5.5-21.8) µg Tl L-1, respectively. A no observable effect concentration (NOEC) of 0.9 µg Tl L-1 for growth, and a NOEC range of 0.9-83 µg Tl L-1 for a variety of reproductive metrics, was measured. A lowest observable effect concentration (LOEC) of 8.8 µg Tl L-1 was determined for the effects of Tl on growth and most of the reproductive endpoints examined. These data indicate that under controlled laboratory conditions D. magna is significantly less sensitive to Tl than the species on which the current Canadian Council of Ministers of the Environment regulatory guideline value of 0.8 µg L-1 is based.

Collective Dynamics of Model Pili-Based Twitcher-Mode Bacilliforms.

Pseudomonas aeruginosa, like many bacilliforms, are not limited only to swimming motility but rather possess many motility strategies. In particular, twitching-mode motility employs hair-like pili to transverse moist surfaces with a jittery irregular crawl. Twitching motility plays a critical role in redistributing cells on surfaces prior to and during colony formation. We combine molecular dynamics and rule-based simulations to study twitching-mode motility of model bacilliforms and show that there is a critical surface coverage fraction at which collective effects arise. Our simulations demonstrate dynamic clustering of twitcher-type bacteria with polydomains of local alignment that exhibit spontaneous correlated motions, similar to rafts in many bacterial communities.

Understanding the effects of hydraulic fracturing flowback and produced water (FPW) to the aquatic invertebrate, Lumbriculus variegatus under various exposure regimes.

Hydraulic fracturing of horizontal wells is a cost effective means for extracting oil and gas from low permeability formations. Hydraulic fracturing often produces considerable volumes of flowback and produced water (FPW). FPW associated with hydraulic fracturing has been shown to be a complex, often brackish mixture containing a variety of anthropogenic and geogenic compounds. In the present study, the risk of FPW releases to aquatic systems was studied using the model benthic invertebrate, Lumbriculus variegatus and field-collected FPW from a fractured well in Alberta. Acute, chronic, and pulse toxicity were evaluated to better understand the implications of accidental FPW releases to aquatic environments. Although L.variegatus is thought to have a high tolerance to many stressors, acute toxicity was significant at low concentrations (i.e. high dilutions) of FPW (48 h LC50: 4-5%). Chronic toxicity (28 d)of FPW in this species was even more pronounced with LC50s (survival/reproduction) and EC50s (total mass) at dilutions as low as 0.22% FPW. Investigations evaluating pulse toxicity (6 h and 48 h exposure) showed a significant amount of latent mortality occurring when compared to the acute results. Additionally, causality in acute and chronic bioassays differed as acute toxicity appeared to be primarily driven by salinity, which was not the case for chronic toxicity, as other stressors appear to be important as well. The findings of this study show the importance of evaluating multiple exposure regimes, the complexity of FPW, and also shows the potential aquatic risk posed by FPW releases.

Non-invasive methods to measure inter-renal function in aquatic salamanders-correlating fecal corticosterone to the environmental and physiologic conditions of captive Necturus.

This study sought to develop non-invasive techniques to monitor glucocorticoids in captive Necturus as a means to correlate inter-renal gland function in relation to environmental and physiological changes. Six individually housed breeding pairs of captive Necturus beyeri were subjected to seasonal changes in water temperature (30°F temperature differential) to stimulate natural breeding, specifically spermatophore deposition and oviposition. An enzyme immunoassay was validated for the measurement of N. beyeri faecal corticosterone metabolites (fCMs) by exhibiting parallelism and accuracy to the standard curve. Longitudinal (December 2016-October 2017) assessment of fCM concentrations and pattern of excretion from samples collected from the six breeding pairs revealed a seasonal inter-renal effect with higher concentrations (P < 0.05) excreted during months (December-March) of the year associated with breeding activity and when water temperatures were lowest. Males from each pair produced spermatophores starting on 08 December 8 2016 and ending on 05 April 2017. Females from four of the six pairs went on to successfully oviposit eggs in mid-late April 2017. One clutch was fertile, and three were non-fertile. No differences (P > 0.05) were detected in fCM concentrations between pairs in which oviposition did or did not occur. In addition, a novel waterborne corticosterone metabolite (wCM) assay was validated to overcome challenges associated with faecal collection in a group-housed amphibian. An adrenocorticotropic hormone (ACTH) challenge performed in an adult male Necturus maculosus resulted in a 50-fold increase in wCM at 4 h post-injection and marked the first demonstration of a waterborne inter-renal response to ACTH in Necturus. This study not only provides insight into inter-renal function in an aquatic salamander that exhibits marked reproductive seasonality but also confirms utility of fCM and wCM measurements as non-invasive means of assessment.

The Effect of Major Ions and Dissolved Organic Matter on Complexation and Toxicity of Dissolved Thallium to Daphnia magna.

Thallium (Tl) is a trace element associated with base metal mining and processing, but little is known regarding how its toxicity is influenced by water chemistry. In the present study, the 48-h median lethal concentration (LC50) of Tl to Daphnia magna was determined in a standard laboratory water, and toxicity was reassessed under conditions of varying cation (Ca2+ , K+ , Na+ ), anion (Cl- , HCO-3 ), and dissolved organic matter (DOM) concentrations. The calculated 48-h LC50 of 1.86 mg Tl/L was consistent with previous work on Tl toxicity to D. magna. At the 48-h LC50 concentration, changes in water chemistry had no statistically significant effect on mortality, although there was a trend toward lower Tl toxicity with elevated water K+ . Test waters containing 10 mM CaCl2 did not support control survival. The measurement of Tl complexation with DOM using asymmetric flow field flow fractionation confirmed the outcomes of biogeochemical speciation modeling: Tl speciation was relatively unaffected by water chemistry, and the majority of Tl remained in the ionic form across all treatments. These data indicate that Tl toxicity is largely independent of speciation, a property that will greatly simplify risk assessments for this metal in freshwaters. Environ Toxicol Chem 2019;38:2472-2479. © 2019 SETAC.

Bioaccumulation of Tl in otoliths of Trout-perch (Percopsis omiscomaycus) from the Athabasca River, upstream and downstream of bitumen mining and upgrading.

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.

The value of gamma camera and computed tomography data set coregistration to assess Lewis Y antigen targeting in small cell lung cancer by (111)Indium-labeled humanized monoclonal antibody 3S193.

AIM: To assess the value of data set coregistration of gamma camera and computed tomography (CT) in the assessment of targeting of humanized monoclonal antibody 3S193 labeled with indium-111 ((111)In-hu3S193) to small cell lung cancer (SCLC). METHODS AND MATERIALS: Ten patients (6 male and 4 female; mean age+/-S.D., 60+/-4 years), from an overall population of 20 patients with SCLCs expressing Lewis Y antigen at immunohistochemical analysis, completed a four weekly injections of (111)In-hu3S193 and underwent gamma camera imaging. All had had, as part of their baseline evaluation, Fluorine18 fluoro-2-deoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT). Two readers in consensus retrospectively coregistered the gamma camera images with the CT component of the FDG PET/CT by automatic or manual alignment. The resulting image sets were visually examined and SCLC lesions targeting at coregistered gamma camera and CT was correlated side-by-side with the (18)F-FDG uptake. RESULTS: A total number of 31 lesions from SCLC with a thoracic (n=13) or extrathoracic location (n=18) were all positive on FDG PET/CT. Coregistration of the gamma camera to the CT demonstrated targeting of antibody to all lesions >2 cm (n=20) and in a few lesions < or =2 cm (n=2), with no visualization of most lesions < or = 2 cm (n=9). No (111)In-hu3S193 uptake in normal tissues was observed. CONCLUSION: Coregistration of antibody gamma camera imaging to FDG PET/CT is feasible and allows valuable assessment of (111)In-hu3S193 antibody targeting to SCLC lesions >2cm, while lesions < or =2 cm reveal a limited targeting.

Targeting Lewis Y (Le(y)) in small cell lung cancer with a humanized monoclonal antibody, hu3S193: a pilot trial testing two dose levels.

INTRODUCTION: Lewis Y (Le(y)) is a blood group antigen with robust expression on the surface of epithelial tumors, including small cell lung cancer (SCLC), making it a potential target for antibody-based immunotherapy. 3S193, an immunoglobulin G3 monoclonal antibody, has demonstrated superior specificity, affinity, and cytotoxicity over other anti-Le(y) antibodies. A phase I trial of humanized 3S193 (hu3S193) with dosing up to 40 mg/m2 demonstrated tumor targeting without serious toxicities or the development of human anti-human antibodies. METHODS: We tested the targeting and pharmacokinetics of hu3S193 in patients with SCLC. Eligibility required progressive SCLC treated with up to three previous chemotherapy regimens, measurable disease not previously irradiated, and tumor samples positive for 3S193 by immunohistochemistry. Patients received four weekly injections of hu3S193, five patients at 10 mg/m2 and five patients at 20 mg/m2. The first and fourth injections were radiolabeled with indium-111 for gamma camera imaging. RESULTS: Of 40 patients screened, 25 of 34 (74%) assessable SCLC tumor samples were 3S193 positive by immunohistochemistry. Ten patients were treated with hu3S193; nine completed all four injections. All fluorodeoxyglucose (FDG)-avid lesions >2 cm were visualized on antibody single-photon emission computed tomography. Some lesions overlying vascular structures could not be visualized. No difference was noted in imaging or pharmacokinetics between the first and fourth injections. Toxicities included grade 2 urticaria (n = 1), grade 1 vomiting (n = 2), and grade 2 hypertension (n = 1) transiently after infusion at the higher dose. CONCLUSIONS: Given the strong tumor targeting, particularly at the higher dose, the favorable toxicity profile, and the potential for immunomodulatory effects, hu3S193 warrants further investigation in SCLC.

Improved tumor imaging and therapy via i.v. IgG-mediated time-sequential modulation of neonatal Fc receptor.

The long plasma half-life of IgG, while allowing for enhanced tumor uptake of tumor-targeted IgG conjugates, also results in increased background activity and normal-tissue toxicity. Therefore, successful therapeutic uses of conjugated antibodies have been limited to the highly sensitive and readily accessible hematopoietic tumors. We report a therapeutic strategy to beneficially alter the pharmacokinetics of IgG antibodies via pharmacological inhibition of the neonatal Fc receptor (FcRn) using high-dose IgG therapy. IgG-treated mice displayed enhanced blood and whole-body clearance of radioactivity, resulting in better tumor-to-blood image contrast and protection of normal tissue from radiation. Tumor uptake and the resultant therapeutic response was unaltered. Furthermore, we demonstrated the use of this approach for imaging of tumors in humans and discuss its potential applications in cancer imaging and therapy. The ability to reduce the serum persistence of conjugated IgG antibodies after their infusion can enhance their therapeutic index, resulting in improved therapeutic and diagnostic efficacy.

Preoperative characterisation of clear-cell renal carcinoma using iodine-124-labelled antibody chimeric G250 (124I-cG250) and PET in patients with renal masses: a phase I trial.

BACKGROUND: Preoperative identification of tumour type could have important implications for the choice of treatment for renal cancers. Antibody cG250 reacts against carbonic anhydrase-IX, which is over-expressed in clear-cell renal carcinomas. We aimed to assess whether iodine-124-labelled antibody chimeric G250 ((124)I-cG250) PET predicts clear-cell renal carcinoma, the most common and aggressive renal tumour. METHODS: 26 patients with renal masses who were scheduled to undergo surgical resection by laparotomy received a single intravenous infusion of 185 MBq/10 mg of (124)I-cG250 over 20 min in this open-label pilot study. Surgery was scheduled 1 week after (124)I-cG250 infusion. PET and CT scanning of the abdomen, including the kidneys, within 3 h before surgery was planned for all patients. The obtained images were graded as positive (defined as a tumour-to-healthy-kidney ratio >3 to 1) or negative for antibody uptake, and the surgeon was informed of the scan results before surgery. After surgery, resected tumours were histopathologically classified as clear-cell renal carcinoma or otherwise. The trial is registered on the clinical trials site of the National Cancer Institute website http://clinicaltrials.gov/ct/show/NCT00199888. FINDINGS: One patient received inactive antibody and was excluded from analysis. 15 of 16 clear-cell carcinomas were identified accurately by antibody PET, and all nine non-clear-cell renal masses were negative for the tracer. The sensitivity of (124)I-cG250 PET for clear-cell kidney carcinoma in this trial was 94% (95% CI 70-100%); the negative predictive value was 90% (55-100%), and specificity and positive predictive accuracy were both 100% (66-100% and 78-100%, respectively). INTERPRETATION: PET with (124)I-cG250 can identify accurately clear-cell renal carcinoma; a negative scan is highly predictive of a less aggressive phenotype. Stratification of patients with renal masses by (124)I-cG250 PET can identify aggressive tumours and help decide treatment.