Elizabeth I. Hamelin

Supplementary Figure S1 shows Pearson correlation coefficients of opioid and nonopioid biomarkers related to opium use among those who use opium by smoking. Colors correspond to the color key on the right, showing the strength and direction of correlation. Squares marked with an “x” show statistically non-significant correlations.

Background:Opium consumption, a recognized carcinogen, is associated with lung cancer. However, the relative contributions of its potentially carcinogenic constituents to lung cancer risk remain unclear. Methods:We conducted a nested case–control study among opium users in the Golestan Cohort Study, including 53 incident lung cancer cases and 53 controls matched on age, sex, tobacco use, place of residence, and enrollment time. Urinary biomarkers of opioids, volatile organic compounds (VOC), polycyclic aromatic hydrocarbons (PAH), cotinine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), and creatinine were measured in baseline samples. Adjusted logistic regression analyses, as well as mediation analyses, were used to evaluate the associations between biomarker concentrations and lung cancer incidence. Results:We found statistically significant associations between 3- and 6-glucuronide of morphine, as well as metabolites of acrylonitrile/ethylene oxide (2-hydroxyethyl mercapturic acid) and propylene oxide (2-hydroxypropyl mercapturic acid), and lung cancer among opium users, especially those who smoked opium. Associations between VOC metabolites and lung cancer were independent of opioid biomarkers, and mediation analysis showed no evidence that VOCs mediate the effects of opioid biomarkers on lung cancer. No associations were observed for PAH biomarkers, cotinine and NNAL. Conclusions:Our findings provide biochemical evidence supporting the association between opium use and lung cancer, highlighting independent mechanisms related to both opioid and non-opioid components of opium, with varying effects by route of use. Impact:Opioids and chemical constituents produced during opium consumption must be considered in cancer risk assessment.See related In the Spotlight, p. 1801

Supplementary Figure S2 shows Pearson correlation coefficients of opioid and nonopioid biomarkers related to opium use among those who use opium by ingestion. Colors correspond to the color key on the right, showing the strength and direction of correlation. Squares marked with an “x” show statistically non-significant correlations.

Supplementary Figure S3 shows Pearson correlation coefficients of opioid and nonopioid biomarkers related to opium use in the total sample. Colors correspond to the color key on the right, showing the strength and direction of correlation. Squares marked with an “x” show statistically non-significant correlations.

Supplementary Table S2 shows Associations between VOC and PAH biomarkers and lung cancer, stratified by routes of opium use

Supplementary Table S3 shows Coefficients from mediation analysis on the indirect effect of VOC metabolites mediating the associations between morphine glucuronides and lung cancer (third tertile compared to first tertile) among people who used opium by smoking

Supplementary Table S1 shows Biomarkers related to opium exposure and their coefficients of variation

Background: Opium consumption, a recognized carcinogen, is associated with lung cancer. However, the relative contributions of its potentially carcinogenic constituents to lung cancer risk remain unclear. Methods: We conducted a nested case–control study among opium users in the Golestan Cohort Study, including 53 incident lung cancer cases and 53 controls matched on age, sex, tobacco use, place of residence, and enrollment time. Urinary biomarkers of opioids, volatile organic compounds (VOC), polycyclic aromatic hydrocarbons (PAH), cotinine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), and creatinine were measured in baseline samples. Adjusted logistic regression analyses, as well as mediation analyses, were used to evaluate the associations between biomarker concentrations and lung cancer incidence. Results: We found statistically significant associations between 3- and 6-glucuronide of morphine, as well as metabolites of acrylonitrile/ethylene oxide (2-hydroxyethyl mercapturic acid) and propylene oxide (2-hydroxypropyl mercapturic acid), and lung cancer among opium users, especially those who smoked opium. Associations between VOC metabolites and lung cancer were independent of opioid biomarkers, and mediation analysis showed no evidence that VOCs mediate the effects of opioid biomarkers on lung cancer. No associations were observed for PAH biomarkers, cotinine and NNAL. Conclusions: Our findings provide biochemical evidence supporting the association between opium use and lung cancer, highlighting independent mechanisms related to both opioid and non-opioid components of opium, with varying effects by route of use. Impact: Opioids and chemical constituents produced during opium consumption must be considered in cancer risk assessment. See related In the Spotlight, p. 1801

Harmful cyanobacterial blooms are becoming more common and persistent around the world. When in bloom, various cyanobacterial strains can produce anatoxins in high concentrations, which, unlike other cyanobacterial toxins, may be present in clear water. Potential human and animal exposures to anatoxins occur mainly through unintentional ingestion of contaminated algal mats and water. To address this public health threat, we developed and validated an LC-MS/MS method to detect anatoxins in human urine to confirm exposures. Pooled urine was fortified with anatoxin-a and dihydroanatoxin at concentrations from 10.0 to 500 ng/mL to create calibrators and quality control samples. Samples were diluted with isotopically labeled anatoxin and solvent prior to LC-MS/MS analysis. This method can accurately quantitate anatoxin-a with inter- and intraday accuracies ranging from 98.5 to 103% and relative standard deviations < 15%, which is within analytical guidelines for mass spectrometry methods. Additionally, this method qualitatively detects a common degradation product of anatoxin, dihydroanatoxin, above 10 ng/mL. We also evaluated a commercial anatoxin-a ELISA kit for potential diagnostic use; however, numerous false positives were detected from unexposed individual human urine samples. In conclusion, we have developed a method to detect anatoxins precisely and accurately in urine samples, addressing a public health area of concern, which can be applied to future exposure events.