Toenail arsenic species and metallome profiles associated with breast, cervical, prostate, and skin cancer prevalence in the Atlantic Partnership for Tomorrow's Health cohort.

Introduction: Chronic exposure to arsenic through drinking water has been linked to several cancers. The metabolism of arsenic is thought to play a key role in arsenic-related carcinogenesis as metabolites of varying toxicity are produced and either stored in or excreted from the body. Atlantic Canada has the highest age-standardized incidence rates of all cancers in the country. This may be due to its high levels of environmental arsenic and the prevalence of unregulated private wells for water consumption. Here, we aimed to characterize the profiles of arsenic species and metallome in the toenails of four cancer groups, compare them to healthy participants (N = 338), and assess potential associations between the profiles with cancer prevalence. Methods: This study employed a case-control design. Toenail samples and questionnaire data from cases (breast, cervical, prostate, and skin cancers) and controls were sourced from the Atlantic Partnership for Tomorrow's Health (PATH) cohort study. The levels of arsenic species were measured using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) paired with High Performance Liquid Chromatography (HPLC) and total concentrations of metallome (23 metals) were determined by ICP-MS separately. Multivariate analyses were conducted to compare cases with controls within each cancer group. Results: Arsenic speciation profiles varied by cancer type and were significantly different between cases and controls in the breast (p = 0.0330), cervical (p = 0.0228), and skin (p = 0.0228) cancer groups. In addition, the profiles of metallome (nine metals) were significantly differentiated in the prostate (p = 0.0244) and skin (p = 0.0321) cancer groups, with higher zinc concentrations among cases compared to controls. Conclusion: History of cancer diagnosis was associated with specific profiles of arsenic species and metallome. Our results indicate that arsenic methylation and zinc levels, as measured in toenails, may be an important biomarker for cancer prevalence. Further research is needed to use toenails as a prognostic measure of arsenic-and other metal-induced cancer.

Arsenic Speciation and Metallomics Profiling of Human Toenails as a Biomarker to Assess Prostate Cancer Cases: Atlantic PATH Cohort Study.

Chronic exposure to inorganic arsenic and trace metals has been linked to prostate cancer, and altered arsenic methylation capacity may have an important role in arsenic carcinogenesis. Biomarkers may be able to elucidate this role. Our objectives were to characterize profiles of arsenic species and metallome in toenails and urine samples, compare profiles between prostate cancer cases and controls, and determine the discriminant ability of toenail and urine biomarkers. Toenail samples (n = 576), urine samples (n = 152), and questionnaire data were sourced from the Atlantic Partnership for Tomorrow's Health (PATH) cohort study. Healthy controls were matched to prostate cancer cases (3:1 ratio) on sex, age, smoking status, and the province of residence. Metallome profiles and proportions of arsenic species were measured in toenail and urine samples. Analysis of covariance (ANCOVA) was used to compare the mean percent monomethylarsonic acid (%MMA), dimethylarsonic acid (%DMA), inorganic arsenic (%iAs), primary methylation index (PMI, MMA/iAs), and secondary methylation index (SMI, DMA/MMA). Multivariate analysis of covariance (MANCOVA) was used to compare selected metal concentrations. Mean %MMA was significantly lower and SMI was significantly higher in toenails from prostate cancer cases compared to controls in unadjusted and adjusted models. Proportions of arsenic species were correlated with total arsenic in toenails. Arsenic speciation in urine was not different between cases and controls, nor were metallome profiles in toenails and urine. Our results indicate that toenails are a viable biomarker for altered arsenic speciation in prostate cancer cases and may have greater utility than urine in this context.

Applying Machine Learning to Arsenic Species and Metallomics Profiles of Toenails to Evaluate Associations of Environmental Arsenic with Incident Cancer Cases.

Chronic exposure to environmental arsenic has been linked to a number of human diseases affecting multiple organ systems, including cancer. The greatest concern for chronic exposure to arsenic is contaminated groundwater used for drinking as it is the main contributor to the amount of arsenic present in the body. An estimated 40% of households in Nova Scotia (Canada) use water from private wells, and there is a concern that exposure to arsenic may be linked to/associated with cancer. In this preliminary study, we are aiming to gain insights into the association of environmental metal's pathogenicity and carcinogenicity with prostate cancer. We use toenails as a novel biomarker for capturing long-term exposure to arsenic, and have performed toxicological analysis to generate data about differential profiles of arsenic species and the metallome (entirety of metals) for both healthy and individuals with a history cancer. We have applied feature selection and machine learning algorithms to arsenic species and metallomics profiles of toenails to investigate the complex association between environmental arsenic (as a carcinogen) and prostate cancer. We present machine learning based models to ultimately predict the association of environmental arsenic exposure in cancer cases.

Toenail speciation biomarkers in arsenic-related disease: a feasibility study for investigating the association between arsenic exposure and chronic disease.

Long-term exposure to environmental arsenic has been associated with many chronic diseases, including several cancers, and diabetes. Urinary studies have implicated arsenic speciation as an important risk factor, however, such associations have not been replicated using toenail samples: a relatively new biosample for estimating long-term internal dose-exposure to arsenic. Despite having several advantages over conventional biosamples such as ease of collection and storage, standard methods for arsenic speciation analysis in toenails have not yet been established. The primary objectives of this study were to 1) establish an analytical method for arsenic speciation analysis in toenails, 2) describe preliminary arsenic speciation profiles of toenail samples from individuals with skin, lung, bladder, and kidney cancer, type II diabetes, and no known disease, and 3) determine if these speciation patterns differ between disease groups to inform the feasibility of subsequent research. A small cross-sectional feasibility study was carried out using 60 toenail samples and baseline questionnaire data from the Atlantic Partnership for Tomorrow's Health (Atlantic PATH) study. Arsenic speciation profiles were determined using high performance liquid chromatography (HPLC) paired with inductively coupled plasma-mass spectrometry (ICP-MS). While no differences in total arsenic were found, arsenic speciation profiles were significantly different between certain cancer groups and the reference group with no known disease. Specifically, the percentage of monomethylarsonic acid (%MMA) was found to be significantly higher in the toenails of individuals with lung cancer and kidney cancer, compared to healthy individuals with similar total arsenic exposure. To the best of our knowledge, this is the first study to describe arsenic speciation patterns in individuals with several arsenic-related diseases using toenails: a convenient, non-invasive, biobankable sample capable of longer-term exposure estimation than conventional biosamples. These preliminary data provide evidence that toenail arsenic speciation patterns differ between groups with arsenic-related disease, and those with no known disease. Toenail arsenic speciation analysis is feasible and could potentially have important implications for research on arsenic-related diseases. Further investigation is warranted and would benefit from including detailed arsenic exposure data to explore the observed heterogeneity in arsenic speciation profiles.

Assessing arsenic in human toenail clippings using portable X-ray fluorescence.

Arsenic is a toxic metalloid which has been associated with a wide range of health effects in humans including skin abnormalities and an elevated risk of skin, bladder, kidney, and lung cancer, diabetes, and cardiovascular disease. The measurement of arsenic concentration in nail clippings is often used in population studies as an indicator of arsenic exposure. Portable X-ray fluorescence (XRF) is an emerging technique for measuring arsenic in nail clippings. In the current study, single toenail clippings from 60 Atlantic Canadian participants were assessed for arsenic using a new portable XRF approach. A mono-energetic portable XRF system using doubly curved crystal optics was used to measure each clipping for a total of 900 s. Energy spectra from each clipping were analyzed for arsenic characteristic X-rays to provide a normalized arsenic signal. The same clippings were then analyzed for arsenic concentration using a "gold standard" method of inductively coupled plasma mass spectrometry (ICP-MS). Nail clipping arsenic concentrations measured by ICP-MS ranged from 0.030 µg/g to 2.57 µg/g, with a median result of 0.14 µg/g. Portable XRF results for arsenic were compared against ICP-MS arsenic concentrations, with a linear equation of best fit determined between the two variables. A correlation coefficient of r = 0.77 was found from the 59 nail clippings returning an ICP-MS arsenic concentration above the limit of quantitation. When the comparison was limited to the 20 clippings having an XRF normalized signal at least twice as large as the associated uncertainty of measurement, the correlation coefficient was r = 0.89. With the selection of an arsenic concentration of 0.1 µg/g as a cut-off value between "exposed" and "non-exposed" individuals, the XRF method provided a test sensitivity of 76% and a specificity of 81%. The corresponding positive predictive value was 88% and the negative predictive value was 65%. The portable XRF technique used in this study shows promise as a means of assessing arsenic concentration in toenail clippings.

Portable X-ray fluorescence of zinc applied to human toenail clippings.

Zinc is an essential trace element in humans. Zinc deficiency can result in a range of serious medical conditions which include effects on growth and development, the immune system, the central nervous system, and the gastrointestinal system. Diagnosis of zinc deficiency is often precluded by the lack of a noninvasive and reliable biomarker. Zinc concentration in nail is considered an emerging biomarker of zinc status in humans. Whether zinc in nail accurately reflects zinc status is beyond the scope of the current study, but is an important research question. The development of a portable method to quickly assess zinc concentration from a single nail clipping could be a useful advance. In this study, single toenail clippings from 60 individuals living in Atlantic Canada were measured for zinc using a portable X-ray fluorescence (XRF) technique. These samples were obtained from the Atlantic PATH cohort, part of the largest chronic disease study ever performed in Canada. Each toenail clipping was measured using three 300 s trials with a mono-energetic portable XRF system. Results were then assessed using two different approaches to the XRF analysis: (1) factory-calibrated zinc concentrations were output from each trial, and (2) energy spectra were analyzed for the characteristic X-rays resulting from zinc. Following the measurement of zinc using the non-destructive portable XRF method, the same clippings were measured for zinc concentration using the "gold standard" technique of inductively coupled plasma-mass spectrometry (ICP-MS). A linear equation of best fit was determined for the relationship between average XRF output zinc concentration and ICP-MS zinc concentration, with a correlation coefficient r = 0.60. Similarly, a linear equation of best fit was found for the relationship between a normalized XRF energy spectrum zinc signal and ICP-MS zinc concentration, with a correlation coefficient r = 0.68. Individual ICP-MS zinc concentrations ranged from 32 µg/g to 140 µg/g, with a population average of 85 µg/g. The results of this study indicate that portable XRF is a sensitive method for the measurement of zinc in a single nail clipping, and provides a reasonable estimation of zinc concentration. Further method development is required before portable XRF be considered a routine alternative to ICP-MS for the assessment of zinc in nail clippings.