Overcoming analytical and preanalytical challenges associated with extragenital home collected STI specimens.

Home sample collection for sexually transmitted infection (STI) screening options can improve access to sexual healthcare across communities. For Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG), genital infections have classically been the focus for remote collection options. However, infections may go undiagnosed if sampling is limited to urogenital sites because some individuals only participate in oral and/or anal intercourse. Here we evaluated samples for CT/NG detection after several pre-analytical collection challenges. A paired provider to self-collection validation was performed on rectal [n = 162; 22 + for CT and 9 + for NG by provider-collected (PC)] and throat (N = 158; 2 + for CT and 11 + for NG by provider-collected) swabs. The positive percent agreement for CT and NG ranged from 90.9% to 100%. The discrepancies were more often positive on self-collected (SC) (n = 9 SC+/PC-; n = 1 PC+/SC-; n = 1 PC+/SC Equiv.; n = 2 PC-/SC Equiv.). An empirical limit of detection (LoD) lower than the manufacturer's claim (0.031 vs 2.5 IFU/mL for CT and 0.063 vs 124.8 CFU/ml for NG, respectively) was used to challenge additional variables. Common hand contaminants, including soap, hand sanitizer, lotion, and sunscreen were added to known positive (3× empirical LoD) or negative samples and did not influence detection. Samples at 2× and 10× the empirical LoD were challenged with extreme temperature cycling and extended room temperature storage. Detection was not affected by these conditions. These results indicate that remote self-collection is an appropriate method of sample acquisition for detecting extragenital CT/NG infections. Additionally, they provide a foundation towards meeting the regulatory standards for commercial testing of home collected extragenital samples. IMPORTANCE: There is a clinical need for expanded extragenital bacterial sexually transmitted infection (STI) testing options, but the current regulatory landscape limits the wide-spread promotion and adoption of such services. Improved access, particularly for the LGBTQ+ community, can be achieved by validating testing for specimens that are self-collected at a remote location and arrive at the laboratory via a postal carrier or other intermediary route. Here we provide valuable data showing that self-collected samples for anal and oropharyngeal STI testing are equally or increasingly sensitive compared with those collected by a provider. We systematically consider the effects of storage time, exposure to temperature extremes, and the addition of common toiletries on results.

Chronic Exposure to a PFAS Mixture Resembling AFFF-Impacted Surface Water Decreases Body Size in Northern Leopard Frogs (Rana pipiens).

Per- and polyfluoroalkyl substances (PFAS) occur in the environment as mixtures, yet mixture toxicity remains poorly understood. Aqueous film-forming foams (AFFFs) are a common source of PFAS. Our objective was to examine chronic effects of a complex PFAS mixture on amphibian growth and development. We tested toxicity of a five-chemical PFAS mixture summing to 10 µg/L and that accounts for >90% of the PFAS in AFFF-affected surface waters: perfluorooctane sulfonate (PFOS, 40%), perfluorohexane sulfonic acid (PFHxS, 30%), perflurooctanoic acid (PFOA, 12.5%), perfluorohexanoic acid (PFHxA, 12.5%), and perfluoropentanoic acid (PFPeA, 5%). We also included treatments to determine whether PFOS drove mixture toxicity and whether PFOS and mixture components act additively. We exposed Northern leopard frog (Rana pipiens) larvae through metamorphosis (∼130 d) in outdoor mesocosms. After 21 days of exposure, the larval body condition fell ∼5% relative to controls in the 4 µg/L PFOS treatment and mixtures lacking PFOS. At metamorphosis, the full 5-component 10 µg/L PFAS mixture reduced mass by 16% relative to controls. We did not observe effects on development. Our results indicate that toxicity of PFOS and other PFAS mixtures typical of AFFF sites act additively and that PFOS is not more inherently toxic than other mixture components.

An Environmentally Relevant Mixture of Perfluorooctanesulfonic Acid and Perfluorohexanesulfonic Acid Does Not Conform to Additivity in Northern Leopard Frogs Exposed Through Metamorphosis.

Per- and polyfluoroalkyl substances (PFAS) are chemicals associated with adverse health effects. At aqueous film-forming foam sites, they occur as mixtures, with perfluorooctanesulfonic acid (PFOS) and perfluorohexanesulfonic acid (PFHxS) commonly co-occurring in the highest concentrations. Although PFOS and PFHxS toxicities have been studied, few studies have tested their potential interaction. Using Rana pipiens, the present study compared toxicities of a 1:1 PFOS:PFHxS mixture to PFOS and PFHxS individually with the prediction that responses would be additive. Gosner stage 25 (GS 25) tadpoles were exposed through metamorphosis (GS 46) to 0.5 and 1 ppb PFOS or PFHxS alone or to a mixture of 0.5 ppb PFOS and 0.5 ppb PFHxS. Tadpoles were weighed and measured (snout-vent length [SVL]) at day 31, metamorphic climax (GS 42), and GS 46. These values were used to calculate the scaled mass index (SMI), a measure of body condition. Body burdens were quantified on day 31 and at GS 46. The PFOS and PFHxS body burdens were elevated relative to controls at GS 46. No effects were observed on survival, SVL, or mass. Single PFAS effects included a 17% reduction in SMI at day 31 (0.5 ppb PFHxS) and a 1.1-day longer metamorphic period (1 ppb PFHxS) relative to controls. Mixture results deviated from additivity-SMIs were higher than expected on day 31 and lower than expected at GS 42. In addition, time to GS 42 in the PFAS mixture exceeded expected additivity by 12 days. Results from a chronic exposure to a 1:1 PFOS:PFHxS mixture resulted in changes in body condition and length of metamorphosis that deviated from additivity. More PFAS mixture toxicity studies conducted at relevant ratios and concentrations are needed. Environ Toxicol Chem 2022;41:3007-3016. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Comparative Toxicity of Aquatic Per- and Polyfluoroalkyl Substance Exposure in Three Species of Amphibians.

Per- and polyfluoroalkyl substances (PFAS) are contaminants of concern due to their widespread occurrence in the environment, persistence, and potential to elicit a range of negative health effects. Per- and polyfluoroalkyl substances are regularly detected in surface waters, but their effects on many aquatic organisms are still poorly understood. Species with thyroid-dependent development, like amphibians, can be especially susceptible to PFAS effects on thyroid hormone regulation. We examined sublethal effects of aquatic exposure to four commonly detected PFAS on larval northern leopard frogs (Rana [Lithobates] pipiens), American toads (Anaxyrus americanus), and eastern tiger salamanders (Ambystoma tigrinum). Animals were exposed for 30 days (frogs and salamanders) or until metamorphosis (toads) to 10, 100, or 1000 µg/L of perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS), or 6:2 fluorotelomer sulfonate (6:2 FTS). We determined that chronic exposure to common PFAS can negatively affect amphibian body condition and development at concentrations as low as 10 µg/L. These effects were highly species dependent, with species having prolonged larval development (frogs and salamanders) being more sensitive to PFAS than more rapidly developing species (toads). Our results demonstrate that some species could experience sublethal effects at sites with surface waters highly affected by PFAS. Our results also indicate that evaluating PFAS toxicity using a single species may not be sufficient for accurate amphibian risk assessment. Future studies are needed to determine whether these differences in susceptibility can be predicted from species' life histories and whether more commonly occurring environmental levels of PFAS could affect amphibians. Environ Toxicol Chem 2022;41:1407-1415. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Anticholinergic burden of patients with dementia attending a Psychiatry of Later Life service.

OBJECTIVES: Older adults with dementia are particularly vulnerable to adverse outcomes resulting from anticholinergic use. We aimed to: (i) Examine the anticholinergic burden of patients with dementia attending a Psychiatry of Later Life (PLL) service (ii) Examine concomitant prescription of acetylcholinesterase inhibitors (AChEIs) and anticholinergics and (iii) Compare the Anticholinergic Cognitive Burden (ACB) scale with a recently published composite list of anticholinergics. METHODS: Retrospective chart review of new referrals with a diagnosis of dementia (n = 66) seen by the PLL service, Tallaght University Hospital, Dublin, Ireland, over a consecutive period of 4 months. RESULTS: The mean ACB score was 2.2 (range = 0-9, SD = 2.1). 37.9% (n = 25) had a clinically significant ACB score (>3) and 42.1% (n = 8) of those taking AChEIs had a clinically significant ACB score. A significantly greater number of medications with anticholinergic activity were identified using the composite list versus the traditional ACB scale (2.3 v.1.5, p = 0.001). CONCLUSIONS: We demonstrated a significant anticholinergic burden amongst patients with dementia attending a specialist PLL service. There was no difference in anticholinergic burden between groups prescribed and not prescribed AChEIs, indicating that these medications are being prescribed without discontinuation of potentially inappropriate medications with anticholinergic activity. The true anticholinergic burden experienced by patients may be underestimated by the use of the ACB score alone, although the clinical significance of this finding is unclear. Calculation of true clinical anticholinergic burden load and its translation to a specific rating scale remains a challenge.

Divergence in heritable life history traits suggests potential for local adaptation and trade-offs associated with a coal ash disposal site.

Globally, human activities have resulted in rapid environmental changes that present unique challenges for wildlife. However, investigations of local adaptation in response to simultaneous exposure to multiple anthropogenic selection pressures are rare and often generate conflicting results. We used an in situ reciprocal transplant design within a quantitative genetic framework to examine how adaptive evolution and phenotypic plasticity contribute to the persistence of an amphibian population inhabiting an environment characterized by high levels of multiple toxic trace elements. We found evidence of phenotypic divergence that is largely consistent with local adaptation to an environment contaminated with multiple chemical stressors, tied to potential trade-offs in the absence of contaminants. Specifically, the population derived from the contaminated environment had a reduced risk of mortality and greater larval growth and in the contaminated environment, relative to offspring from the naïve population. Further, while survival in the uncontaminated environment was not compromised in offspring from the contaminant-exposed population, they did show delayed development and reduced growth rates over larval development, relative to the naïve population. We found no evidence of reduced additive genetic variation in the contaminant-exposed population, suggesting long-term selection in a novel environment has not reduced the evolutionary potential of that population. We also saw little evidence that past selection in the ASH environment had reduced trait plasticity in the resident population. Maternal effects were prominent in early development, but we did not detect any trends suggesting these effects were associated with the maternal transfer of toxic trace elements. Our results demonstrate the potential for adaptation to multiple contaminants in a wild amphibian population, which may have facilitated long-term persistence in a heavily impacted environment.

Does instillation of lidocaine gel following flexible cystoscopy decrease the severity of post procedure symptoms? A randomised controlled trial assessing the efficacy of lidocaine gel post flexible cystoscopy.

OBJECTIVE: To assess whether instillation of lidocaine gel both before and after flexible cystoscopy is more effective at reducing post procedural symptoms than instillation of lidocaine gel pre flexible cystoscopy alone. We hypothesise that inadequate urethral dwell time and dilution of lidocaine gel by the irrigation fluid during flexible cystoscopy limits its anaesthetic efficacy. Only one other study has attempted to reduce bothersome urinary symptoms through an intervention after flexible cystoscopy. METHODS: This was a randomised controlled trial in which patients were randomised 1:1 to receive lidocaine gel pre and post flexible cystoscopy (treatment) or lidocaine gel pre flexible cystoscopy only (control). Patient-reported outcome measures were used to assess symptoms and quality of life prior to cystoscopy, on day 2 and day 7 post cystoscopy. RESULT: Fifty patients were divided equally between the treatment and control groups. There were no significant differences in baseline characteristics between the groups (p = 1.000). An overall symptoms variable was measured, though no significant difference was found in the distribution of responses between the groups at baseline, 2 or 7 days after the flexible cystoscopy (p = 0.423, 0.651,0.735). In the treatment group, 1 patient (4.0%) presented to a doctor for review following flexible cystoscopy, and 4 patients (16.0%) presented in the control group (p = 0.349). CONCLUSION: Initial study results suggest that post-operative lidocaine does not significantly limit the exacerbation of urinary symptoms following flexible cystoscopy; however, our results are not powered to detect a small difference. We do not recommend a change in practice based on our results.

Dietary exposure and accumulation of per- and polyfluoroalkyl substances alters growth and reduces body condition of post-metamorphic salamanders.

Per- and polyfluoroalkyl substances (PFAS) are contaminants of concern due to their persistence, potential to bioaccumulate, and toxicity. While dietary exposure is the primary route of exposure for terrestrial species, data on dietary PFAS uptake and adverse effects are largely restricted to mammals. As such, substantial data gaps exist that hinder ecological risk assessment, including environmentally relevant exposure levels and taxa. Using a 30-d laboratory experiment, we examined the effects of dietary PFAS-exposure on post-metamorphic tiger salamanders (Ambystoma tigrinum). We fed salamanders crickets exposed to perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS), or 6:2 fluorotelomer sulfonate (6:2 FTS) at low (<1.0), medium (2-5), or high (16-62) ng PFAS/g/d (wet weight) dose rates. We found that only PFOS resulted in substantial biomagnification. Despite limited evidence for biomagnification, PFAS altered growth and generally reduced body condition. Salamanders with the highest burdens of PFOS grew less and had lower body conditions, while burdens of PFHxS and PFOA were only associated with reduced growth. There was no evidence that environmentally relevant doses of PFAS increase liver size in salamanders. Our results demonstrate that dietary exposure and accumulation of PFAS can impact fitness-related traits in amphibians and contribute to trophic transfer in terrestrial food webs.

Perfluoroalkyl Substances Increase Susceptibility of Northern Leopard Frog Tadpoles to Trematode Infection.

Per/polyfluoroalkyl substances (PFAS) are contaminants of emerging concern that can impair immune function, yet few studies have tested whether exposure increases infection risk. Using laboratory experiments, we found that exposure to 10 ppb of perfluorohexanesulfonic acid increased trematode (Echinoparyphium lineage 3) infections in larval northern leopard frogs (Lithobates pipiens). However, there was no effect of perfluorooctanesulfonic acid. Our results demonstrate that PFAS can potentially enhance infection risk in natural systems. Environ Toxicol Chem 2021;40:689-694. © 2020 SETAC.

Bladder cancer in Ireland: a 21-year review.

INTRODUCTION: Bladder carcinoma is one of the most commonly diagnosed cancers worldwide; however, various studies have reported on marked international variation in incidence and mortality rates. The aim of this study was to report national figures on incidence, survival and risk factors of bladder cancer. This retrospective review is the first to offer a comprehensive analysis of incidence rates, mortality and potential risk factors in the Irish population from 1994 to 2015. METHODS: This was a retrospective study of incidence and survival trends in bladder cancer in the Irish population from 1994 to 2014 based on data from the National Cancer Registry of Ireland (NCRI). Patient demographics, pathology and survival data, as well as potential contributing factors, were studied. RESULTS: A total of 9656 cases of bladder cancer were diagnosed from 1994 to 2015, with majority of cases occurring in males (71%). Survival decreased with increasing age, higher grade and higher stage. Bladder cancer is more common in those with a smoking history and with a higher deprivation score. Urothelial cell carcinoma was the most common pathology (95%), with the majority of patients being stage 1 or 2 (44.7%). CONCLUSION: The overall incidence of bladder cancer in Ireland has fallen, in line with reducing smoking rates. Our data reflects international trends in incidence, survival and contributing risk factors.

Chronic Per-/Polyfluoroalkyl Substance Exposure Under Environmentally Relevant Conditions Delays Development in Northern Leopard Frog (Rana pipiens) Larvae.

Per-/polyfluoroalkyl substances (PFAS) are pervasive in aquatic systems globally and capable of causing detrimental effects on human and wildlife health. However, most studies are conducted under artificial conditions that are not representative of environmental exposures. Environmental exposures are characterized by multiple routes of exposure, low aquatic PFAS levels, and greater environmental variability than laboratory tests. Determining whether these factors influence toxicity is critical for understanding the effects of PFAS on aquatic life, including amphibians. Our goal was to assess the impact of PFAS on an amphibian under semirealistic conditions. We reared northern leopard frog (Rana pipiens) larvae in outdoor mesocosms containing sediment spiked to low, medium, and high levels (nominally 10, 100, or 1000 ppb dry wt) of perfluorooctanesulfonic acid (PFOS) or perfluorooctanoic acid (PFOA) for 30 d. Larvae in all PFOS treatments and the medium-PFOA treatment were approximately 1.5 Gosner stages less developed than control animals after 30 d. Notably, these developmental delays were observed at PFOS concentrations in the water as low as 0.06 ppb, which is considerably lower than levels associated with developmental effects in laboratory studies. Our results suggest that deriving toxicity values from laboratory studies examining aquatic exposure only may underestimate the effects of environmental PFAS exposure. Environ Toxicol Chem 2021;40:711-716. © 2020 SETAC.

Sublethal Effects of Dermal Exposure to Poly- and Perfluoroalkyl Substances on Postmetamorphic Amphibians.

Studies of the toxicity of poly- and perfluoroalkyl substances (PFAS) on amphibians, especially after metamorphosis, are limited. We examined effects of dermal PFAS exposure (30 d) on survival and growth of juvenile American toads (Anaxyrus americanus), eastern tiger salamanders (Ambystoma tigrinum), and northern leopard frogs (Rana pipiens). Chemicals included perfluorooctanoic acid, perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS), and 6:2 fluorotelomer sulfonate (6:2 FTS) at 0, 80, 800, or 8000 ppb on a moss dry weight basis. Exposure to PFAS influenced final snout-vent length (SVL) and scaled mass index (SMI), a measure of relative body condition. Observed effects depended on species and chemical, but not concentration. Anurans exposed to PFOS, PFHxS (frogs only), and 6:2 FTS demonstrated reduced SVL versus controls, whereas salamanders exposed to 6:2 FTS showed increased SVL. Frogs exposed to PFHxS and 6:2 FTS and toads exposed to PFOS had increased SMI compared to controls; salamanders did not demonstrate effects. Concentrations of 6:2 FTS in substrate decreased substantially by 30 d, likely driven by microbial action. Perfluorooctane sulfonate had notable biota-sediment accumulation factors, but was still <1. Although a no-observable-effect concentration could not generally be determined, the lowest-observable-effect concentration was 50 to 120 ppb. Survival was not affected. The present study demonstrates that PFAS bioaccumulation from dermal exposures and sublethal effects are dependent on species, chemical, and focal trait. Environ Toxicol Chem 2021;40:717-726. © 2020 SETAC.

Single and mixture per- and polyfluoroalkyl substances accumulate in developing Northern leopard frog brains and produce complex neurotransmission alterations.

Per- and polyfluoroalkyl substances (PFAS) are present in water and >99% of human serum. They are found in brains of wildlife; however, little is known about effects on the developing brain. To determine the effects of PFAS on brain and cardiac innervation, we conducted an outdoor mesocosm experiment with Northern leopard frog larvae (Rana pipiens) exposed to control, 10 ppb perfluorooctane sulfonate (PFOS), or a PFAS mixture totaling 10 ppb that mimicked aqueous film forming foam-impacted surface water (4 ppb PFOS, 3 ppb perfluorohexane sulfonate, 1.25 ppb perfluorooctanoate, 1.25 ppb perfluorohexanoate, and 0.5 ppb perfluoro-n-pentanoate). Water was spiked with PFAS and 25 larvae (Gosner stage (GS) 25) added to each mesocosm (n = 4 mesocosms per treatment). After 30 days, we harvested eight brains per mesocosm and remaining larvae developed to GS 46 (i.e. metamorphosis) before brains and hearts were collected. Weight, length, GS, and time to metamorphosis were recorded. Brain concentrations of all five PFAS were quantified using LC/MS/MS. Dopamine and metabolites, serotonin and its metabolite, norepinephrine, γ-aminobutyric acid, and glutamate were quantified using High Performance Liquid Chromatography with electrochemical detection while acetylcholine and acetylcholinesterase activity were quantified with the Invitrogen Amplex Red Acetylcholine Assay. PFOS accumulated in the brain time- and dose-dependently. After 30 days, the mixture decreased serotonin while both PFAS treatments decreased glutamate. Interestingly, acetylcholine increased in PFAS treatments at GS 46. This research shows that developmental environmentally relevant exposure to PFAS changes neurotransmitters, especially acetylcholine.

Acute and chronic effects of perfluoroalkyl substance mixtures on larval American bullfrogs (Rana catesbeiana).

Discovery of elevated concentrations of perfluoroalkyl substances (PFAS) in ground and surface waters globally has heightened concern over their potential adverse health effects. The effects of PFAS are known largely from acute toxicity studies of single PFAS compounds in model organisms, while little is understood concerning effects of mixtures on wildlife. To address this gap, we examined the acute and chronic effects of two of the most common PFAS (perfluorooctanesulfonic acid [PFOS] and perfluorooctanoic acid [PFOA]) and their mixtures on survival, growth, and development of American bullfrog (Rana catesbeiana) tadpoles. In 96 h acute toxicity tests, PFOS was 10X more toxic than PFOA and effects of the two chemicals in combination appeared additive. The effects of PFOS, PFOA, and their interaction varied by the sublethal endpoint under consideration in a 72 d exposure. Effects of PFAS on tadpole mass and developmental stage were largely driven by PFOS and there was no evidence of interactions suggesting deviations from additivity. However, for snout-vent length, reductions in length in mixture treatments were greater than expected based on the effects of the two chemicals independently (i.e. non-additivity). Further, effects on snout-vent length in single chemical exposures were only observed with PFOA. Our results highlight the importance of assessing combined effects of PFAS co-occurring in the environment and suggest caution in extrapolating the effects of acute toxicity studies to more environmentally relevant exposures. Future studies examining effects of environmentally relevant mixtures on wildlife will be essential for effective environmental risk assessment and management.

Developmental exposure to perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) selectively decreases brain dopamine levels in Northern leopard frogs.

Per- and polyfluoroalkyl substances (PFAS) are synthetic compounds that are a major public health concern due to widespread use, long environmental and biological half-lives, detection in most human plasma samples, and links to multiple adverse health outcomes. The literature suggests that some PFAS may be neurotoxic. However, there are major gaps in the literature with respect to how environmentally-relevant doses during development may influence the nervous system. To address this gap, we utilized a sentinel species, Northern leopard frogs (Lithobates pipiens) to determine the effects of developmental exposure to environmentally relevant perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) on major neurotransmitter systems. Frog larvae at Gosner stage 25 were exposed to 10, 100, or 1000 ppb PFOS or PFOA for 30 days before neurochemical analysis. High performance liquid chromatography (HPLC) with electrochemical detection or fluorescent detection assays was used to measure neurotransmitter levels, which were normalized to protein levels in each sample. Dopamine (DA) decreased significantly in the brains of frogs treated with PFOA (1000 ppb) and PFOS (100 and 1000 ppb). Significant increases in DA turnover also resulted from PFOA and PFOS treatment. Neither PFOS, nor PFOA produced detectable alterations in serotonin (nor its metabolite), norepinephrine, gamma-amino butyric acid (GABA), glutamate, or acetylcholine. PFAS body burdens showed that PFOS accumulated relative to dose, while PFOA did not. These data suggest that DArgic neurotransmission is selectively affected in developmentally exposed amphibians and that PFAS should be evaluated for a potential role in diseases that target the DA system.

Variation in metal tolerance associated with population exposure history in Southern toads (Anaxyrus terrestris).

Human activities have radically shaped the global landscape, affecting the structure and function of ecosystems. Habitat loss is one of the most visible changes to the landscape and a primary driver of species declines; however, anthropogenic environmental contamination also threatens population persistence, but is not as readily observed. Aquatic organisms are especially susceptible to chemical perturbations, which can negatively impact survival and fitness related traits. Some populations have evolved tolerance to chemical stressors, which could mitigate the consequences associated with contamination. Amphibians are experiencing global declines due to multiple stressors and are particularly at risk to aquatic chemical stressors due to their permeable skin and reliance on wetlands for reproduction and larval development. However, amphibians also have substantial plasticity in response to environmental variation. We designed our study to examine whether tolerance to heavy metals is greater in Southern toad (Anaxyrus terrestris) larvae from wetlands with a history of contamination. Considering many of the most common trace elements elicit acute toxicity by disrupting osmotic- and ionic-regulation, we hypothesized that alterations to these aspects of physiology resulting from multigenerational exposure to trace element mixtures would be the most likely routes by which tolerance would evolve. We used copper (Cu) as a proxy for heavy metal exposure because it is a widely distributed aquatic stressor known to cause osmotic stress that can also cause mortality at levels commonly encountered in the environment. We found considerable within and among population variation in Cu tolerance, as measured by time to death. Larvae from populations living in sites contaminated with mixtures of heavy metals associated with coal fly ash were no more tolerant to Cu than those from reference sites. However, larvae from a population inhabiting a constructed wetland complex with high Cu levels were significantly more tolerant; having half the risk of mortality as reference animals. This wetland complex was created < 20 years ago, thus if elevated Cu tolerance in this population is due to selection in the aquatic habitat, such adaptation may occur rapidly (i.e. ∼10 generation). Our results provide evidence that amphibians may be able to evolve tolerance in response to trace element contamination, though such tolerance may be specific to the combination of contaminants present.

Primary Renal Carcinoid - A Case Report.

Carcinoid tumours in the abdomen are uncommon, but typically occur in the gastrointestinal tract. Primary renal carcinoid is an extremely rare tumour, poorly described in the literature. We describe an unusual case where an atypical renal mass on imaging led to a preoperative diagnosis of renal carcinoid on imaging guiding biopsy.