Exposure to benzotriazoles and benzothiazoles in Czech male population and its associations with biomarkers of liver function, serum lipids and oxidative stress.

INTRODUCTION: Benzotriazoles and benzothiazoles (BTs) are high-production volume chemicals as well as widely distributed emerging pollutants with potential health risk. However, information about human exposure to BTs and associated health outcomes is limited. OBJECTIVE: We aimed to characterise exposure to BTs among Czech men, including possible occupational exposure among firefighters, its predictors, and its associations with liver function, serum lipids and oxidative stress. METHODS: 165 participants (including 110 firefighters) provided urine and blood samples that were used to quantify the urinary levels of 8 BTs (high-performance liquid chromatography-tandem mass spectrometry), and 4 liver enzymes, cholesterol, low-density lipoprotein, and 8-hydroxy-2'-deoxyguanosine. Linear regression was used to assess associations with population characteristics and biomarkers of liver function, serum lipids and oxidative stress. Regression models were adjusted for potential confounding variables and false discovery rate procedure was applied to account for multiplicity. RESULTS: The BTs ranged from undetected up to 46.8 ng/mL. 2-hydroxy-benzothiazole was the most predominant compound (detection frequency 83%; median 1.95 ng/mL). 1-methyl-benzotriazole (1M-BTR) was measured in human samples for the first time, with a detection frequency 77% and median 1.75 ng/mL. Professional firefighters had lower urinary 1M-BTR compared to non-firefighters. Urinary 1M-BTR was associated with levels of γ-glutamyl transferase (ß = - 17.54%; 95% CI: - 26.127, - 7.962). CONCLUSION: This is the first study to investigate BT exposure in Central Europe, including potentially exposed firefighters. The findings showed a high prevalence of BTs in the study population, the relevance of 1M-BTR as a new biomarker of exposure, and an urgent need for further research into associated adverse health outcomes.

Countrywide analysis of heat- and cold-related mortality trends in the Czech Republic: growing inequalities under recent climate warming.

BACKGROUND: Only little is known about trends in temperature-mortality associations among the most vulnerable subgroups, especially in the areas of central and eastern Europe, which are considered major climatic hotspots in terms of heatwave exposure. Thus, we aimed to assess trends in temperature-related mortality in the Czech Republic by sex, age and cause of death, and to quantify the temporal evolution of possible inequalities. METHODS: We collected daily time series of all-cause (1987-2019) and cause-specific (1994-2019) mortality by sex and age category, and population-weighted daily mean 2-metre temperatures for each region of the Czech Republic. We applied a quasi-Poisson regression model to estimate the trends in region-specific temperature-mortality associations, with distributed lag non-linear models and multivariate random-effects meta-analysis to derive average associations across the country. We then calculated mortality attributable to non-optimal temperatures and implemented the indicator of sex- and age-dependent inequalities. RESULTS: We observed a similar risk of mortality due to cold temperatures for men and women. Conversely, for warm temperatures, a higher risk was observed for women. Results by age showed a clear pattern of increasing risk due to non-optimum temperatures with increasing age category. The relative risk (RR) related to cold was considerably attenuated in most of the studied subgroups during the study period, whereas an increase in the RR associated with heat was seen in the overall population, in women, in the age category 90+ years and with respect to respiratory causes. Moreover, underlying sex- and age-dependent inequalities experienced substantial growth. CONCLUSIONS: Our findings suggest ongoing adaptation to cold temperatures. Mal/adaptation to hot temperatures occurred unequally among population subgroups and resulted in growing inequalities between the sexes and among age categories.

Current-use pesticide exposure pathways in Czech adults and children from the CELSPAC-SPECIMEn cohort.

INTRODUCTION: In this study, we aimed to characterise exposure to pyrethroids, organophosphates, and tebuconazole through multiple pathways in 110 parent-child pairs participating in the CELSPAC-SPECIMEn study. METHODS: First, we estimated the daily intake (EDI) of pesticides based on measured urinary metabolites. Second, we compared EDI with estimated pesticide intake from food. We used multiple linear regression to identify the main predictors of urinary pesticide concentrations. We also assessed the relationship between urinary pesticide concentrations and organic and non-organic food consumption while controlling for a range of factors. Finally, we employed a model to estimate inhalation and dermal exposure due to spray drift and volatilization after assuming pesticide application in crop fields. RESULTS: EDI was often higher in children in comparison to adults, especially in the winter season. A comparison of food intake estimates and EDI suggested diet as a critical pathway of tebuconazole exposure, less so in the case of organophosphates. Regression models showed that consumption per g of peaches/apricots was associated with an increase of 0.37% CI [0.23% to 0.51%] in urinary tebuconazole metabolite concentrations. Consumption of white bread was associated with an increase of 0.21% CI [0.08% to 0.35%], and consumption of organic strawberries was inversely associated (-61.52% CI [-79.34% to -28.32%]), with urinary pyrethroid metabolite concentrations. Inhalation and dermal exposure seemed to represent a relatively small contribution to pesticide exposure as compared to dietary intake. CONCLUSION: In our study population, findings indicate diet plays a significant role in exposure to the analysed pesticides. We found an influence of potential exposure due to spray drift and volatilization among the subpopulation residing near presumably sprayed crop fields to be minimal in comparison. However, the lack of data indicating actual spraying occurred during the critical 24-hour period prior to urine sample collection could be a significant contributing factor.

Rapid extraction and analysis of oxidative stress and DNA damage biomarker 8-hydroxy-2'-deoxyguanosine (8-OHdG) in urine: Application to a study with pregnant women.

Oxidative stress is an important toxicity and genotoxicity mechanism of many chronic adverse health outcomes. This study developed a sensitive extraction method for urine matrix (based on lyophilization, without the need for pre-cleaning by solid phase extraction), coupled to LC-MS/MS analysis of the biomarker 8-hydroxy-2'-deoxyguanosine (8-OHdG). The methodology was validated in urine samples from a cohort of Spanish pregnant women collected during the first, second and third trimester of pregnancy, and urine samples collected within 24 h after delivery (n = 85). A detection and quantification limit of 0.01 and 0.05 µg/L, respectively, were established. The median 8-OHdG concentration was 2.18 µg/L (range 0.33-7.79); and the corresponding creatinine-adjusted concentrations ranged from 1.04 to 13.12 with median of 4.48 µg 8-OHdG/g creatinine. The concentrations of non-adjusted 8-OHdG significantly decreased (p < 0.05) in the 3rd trimester and post-delivery urine samples when compared to the 1st trimester levels. 8-OHdG concentrations were further studied in placenta samples matching the same urine samples (n = 26), with a median value of 1.3 ng 8-OHdG/g of tissue. Placental 8-OHdG concentrations were correlated with urinary levels of non-adjusted 8-OHdG in the 3rd trimester. Considering the small cohort size, results must be interpreted with caution, however statistical analyses revealed elevated urinary non-adjusted 8-OHdG levels in the 1st trimester of mothers that delivered boys compared to those who delivered girls (p < 0.01). Increased urinary non-adjusted 8-OHdG concentrations at the time of delivery were significantly associated with clinical records (any type of clinical record during pregnancy; p < 0.05). The novel extraction and analytical method for the assessment of 8-OHdG is applicable for sensitive analysis of multiple analytes or biomarkers in urine matrix. This method could also be applied for other matrices such as blood or tissues. Our findings show that 8-OHdG in urine of pregnant women could predict oxidative stress in placenta and can be related to characteristics such as maternal obesity, mode of delivery and newborn sex.

Effects of pesticide exposure on oxidative stress and DNA methylation urinary biomarkers in Czech adults and children from the CELSPAC-SPECIMEn cohort.

Current-use pesticide (CUP) exposure occurs mainly through diet and environmental application in both agricultural and urban settings. While pesticide exposure has been associated with many adverse health outcomes, the intermediary molecular mechanisms are still not completely elucidated. Among others, their roles in epigenetics (DNA methylation) and DNA damage due to oxidative stress are presumed. Scientific evidence on urinary biomarkers of such body response in general population is limited, especially in children. A total of 440 urine samples (n = 110 parent-child pairs) were collected during the winter and summer seasons in order to describe levels of overall DNA methylation (5-mC, 5-mdC, 5-hmdC, 7-mG, 3-mA) and oxidative stress (8-OHdG) biomarkers and investigate their possible associations with metabolites of pyrethroids (3-PBA, t/c-DCCA), chlorpyrifos (TCPY), and tebuconazole (TEB-OH). Linear mixed-effects models accounting for intraindividual and intrahousehold correlations were utilized. We applied false discovery rate procedure to account for multiplicity and adjusted for potential confounding variables. Higher urinary levels of most biological response biomarkers were measured in winter samples. In adjusted repeated measures models, interquartile range (IQR) increases in pyrethroid metabolites were associated with higher oxidative stress. t/c-DCCA and TCPY were associated with higher urinary levels of cytosine methylation biomarkers (5-mC and/or 5-mdC). The most robust association was observed for tebuconazole metabolite with 3-mA (-15.1% change per IQR increase, 95% CI = -23.6, -5.69) suggesting a role of this pesticide in reduced demethylation processes through possible DNA glycosylase inhibition. Our results indicate an urgent need to extend the range of analyzed environmental chemicals such as azole pesticides (e.g. prothioconazole) in human biomonitoring studies. This is the first study to report urinary DNA methylation biomarkers in children and associations between CUP metabolites and a comprehensive set of biomarkers including methylated and oxidized DNA alterations. Observed associations warrant further large-scale research of these biomarkers and environmental pollutants including CUPs.

City-scale assessment of long-term air quality impacts on the respiratory and cardiovascular health.

Background: The impact of the urban environment on human health is a contemporary subject of environmental research. Air pollution is often considered a leading environmental driver. However, a plethora of other factors within the urban exposome may be involved. At the same time, the resolution of spatial data is also an important facet to consider. Generally, systematic tools for accurate health risk assessment in the urban environment are missing or are not implemented. Methods: The long-term impact of air quality (PM10, PM2.5, NO2, benzene, and SO2) on respiratory and cardiovascular health was assessed with a log-linear model. We used the most accurate health data in high city scale spatial resolution over the period 2010 to 2018. Selected external exposome parameters were also included in the analysis. Results: Statistically significant associations between air pollution and the health of the urban population were found. The strongest association was between benzene and the incidence of bronchitis in the adult population [RR 1.552 95% CI (1.415-1.704) per 0.5 µg/m3 change in benzene concentration]. A similar relation was observed between NO2 and the same health condition [RR 1.483 95% CI (1.227-1.792) per 8.9 µg/m3 of change in NO2]. Other weaker associations were also found between asthma in children and PMs, NO2, or benzene. Cardiovascular-related hospitalizations in the general population were linked with NO2 [RR 1.218 95% CI (1.119-1.325) per 9.7 µg/m3 change in NO2]. The remaining pollutants were slightly less but still significantly associated with cardiovascular-related hospitalizations. Conclusion: Our findings are mostly highly statistically significant (p ≤ 0.001) and are in line with current literature on the adverse effects of air pollution on the human population. The results highlight the need for continual improvements in air quality. We propose the implementation of this approach as a systematic tool for the investigation of possible health risks over a long period of time. However, further research involving other variables is an essential step toward understanding the complex urban exposome and its implications for human health. An increase in data spatial resolution is especially important in this respect as well as for improving city health risk management.

Pesticide exposure among Czech adults and children from the CELSPAC-SPECIMEn cohort: Urinary biomarker levels and associated health risks.

Current-use pesticides (CUP) are extensively applied in both agricultural and urban settings. Exposure occurs mainly via the dietary pathway; however, other pathways such as inhalation or skin contact are also important. In this study, urinary levels of 12 CUP metabolites were investigated among 110 parent-child pairs during two seasons of 2020. Metabolites of pyrethroids (3-PBA, t/c-DCCA), chlorpyrifos (TCPY), and tebuconazole (TEB-OH) were detected in more than 60% of the samples. Chlorpyrifos metabolite was found at the highest concentration and tebuconazole was detected in almost all samples. CUP urinary metabolite levels were significantly higher in children in comparison to adults, except for tebuconazole, which was similar in both groups. In children, winter samples had significantly higher concentrations of pyrethroid and chlorpyrifos metabolites in comparison to the summer samples, but in adults, only chlorpyrifos metabolite concentrations were higher in the winter. No association between CUP urinary metabolite levels and proximity/surface of agricultural areas around residences was observed. Based on our findings, we suspect that CUP exposure is mainly driven by diet and that the effect of environmental exposure is less significant. Daily Intakes were estimated with three possible scenarios considering the amount of the metabolite excreted in urine and were compared to Acceptable Daily Intake values. Using a realistic scenario, exposure to chlorpyrifos exhibited the highest health risk, but still within a safe level. The Acceptable Daily Intake was exceeded only in one child in the case of cypermethrin. The cumulative risk assessment of pesticide mixtures having an effect on the nervous system, based on the total margin of exposure calculations, did not indicate any risk. The overall risk associated with pesticide exposure in the observed population was low. However, the risk observed using the worst-case scenario suggests the need for continuous evaluation of human exposure to such compounds, especially in children.

Lifetime Carcinogenic Risk Proportions from Inhalation Exposures in Industrial and Non-Industrial Regions.

The aim of this work was to estimate the share of selected significant risk factors for respiratory cancer in the overall incidence of this disease and their comparison in two environmentally different burdened regions. A combination of a longitudinal cross-sectional population study with a US EPA health risk assessment methodology was used. The result of this procedure is the expression of lifelong carcinogenic risks and their contribution in the overall incidence of the disease. Compared to exposures to benzo[a]pyrene in the air and fibrogenic dust in the working air, several orders of magnitude higher share of the total incidence of respiratory cancer was found in radon exposures, for women 60% in the industrial area, respectively 100% in the non-industrial area, for men 24%, respectively 15%. The share of risks in workers exposed to fibrogenic dust was found to be 0.35% in the industrial area. For benzo[a]pyrene, the share of risks was below 1% and the share of other risk factors was in the monitored areas was up to 85%. The most significant share in the development of respiratory cancer in both monitored areas is represented by radon for women and other risk factors for men.

Biomonitoring of PFOA, PFOS and PFNA in human milk from Czech Republic, time trends and estimation of infant's daily intake.

BACKGROUND: Perfluoralkylated substances (PFASs) are persistent and bioaccumulative environmental contaminants. They are included on the list of emergent compounds monitored in the frame of HBM4EU project. OBJECTIVES: To analyze PFASs levels in human milk samples collected in the period 2006 through 2017, to follow their time trends, to assess the PFASs exposure in breastfed infants, to calculate the daily intake of PFASs and to compare it with the tolerable daily/weekly) intakes and to quantify risk from exposure using the hazard quotient and hazard index approach. MATERIAL AND METHODS: A broad spectrum of PFASs were analyzed by means of UHPLC-MS/MS in primipara human milk samples collected in four consecutive time periods 2006, 2010/11, 2014, and 2017; N = 46, 183, 164 and 232, respectively. Mothers living in urban and suburban residences were recruited after their delivery at maternity hospitals, and milk samples were taken within 2 and 8 weeks after delivery. The questionnaire was focused on possible sources of exposure, dietary habits and lifestyle. RESULTS: Only perfluorooctane sulfonate (PFOS) and perfluorooctanoid acid PFOA (in 2017, also perfluorononanoic acid (PFNA)) were quantified in more than 90% of analyzed human milk samples. In all sampling periods, the levels of PFOA were higher than those of PFOS (p < 0.05). A significant downward temporal trend (p < 0.001) was observed for both PFOA and PFOS levels. The median concentrations in sampling years 2006, 2010/11, 2014, and 2017 were 0.075, 0.059, 0.035, and 0.023 ng/mL for PFOA and 0.045, 0.031, 0.029, and 0.020 ng/mL for PFOS, respectively. In 2017, PFNA was also quantified in 99% of samples with the median concentration of 0.007 ng/mL. The levels of PFASs correlated with maternal sea fish consumption. No maternal age-related relationship was observed. Using the tolerably daily intake (TDI) values for PFOS and PFOA set by the European Food Safety Authority (EFSA) in 2008, the calculated daily intakes from breastfeeding were clearly below these limits. Using the new, more conservative EFSA Provisional Tolerably Weekly Intake (PTWI) values set in 2018, we demonstrated a considerable exceedance of PTWI, with a hazard index above 1. CONCLUSION: Significant time-related decreasing trends in the PFOS and PFOA levels in human milk were observed. Nevertheless, the body burden of infants from breastfeeding might pose an enhanced health risk to infants when the current PTWI values are applied. These findings strongly support the present EU efforts to phase out PFOA, its salts and PFOA related compounds. Since PFOS exposure there has still been widely detected despite PFOS usage reduction measures, the major exposure routes should be further monitored and, if possible, eliminated.

Exposure to Mixed Phthalates in Czech Preschool and School Children.

Knowledge of population exposure to phthalates based on the urinary metabolite levels is of the highest importance for health risk assessment. Such data are scarce in the Czech population. In the study conducted in 2016, six urinary phthalate metabolites were analysed in a total of 370 first morning urine samples of healthy children aged 5 and 9 years, namely mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl) phthalate (5oxo-MEHP), mono-benzyl phthalate (MBzP), mono-iso-butyl phthalate (MiBP), and mono-n-butyl phthalate (MnBP). The two latter mono-butyl phthalate isoforms dominated among all samples with geometric means of 63.0 µg/L (MnBP) and 44.1 µg/L (MiBP), followed by 5OH-MEHP (20.6 µg/L), 5oxo-MEHP (12.9 µg/L), MBzP (3.65 µg/L), and MEHP (2.31 µg/L). Daily intake (DI) of the parent phthalates was estimated using the creatinine-based model. The highest DI values were found for di-n-butyl phthalate (DnBP) (median 2.5 µg/kg bw/day; 95th percentile 7.8 µg/kg bw/day) and di-2-ethylhexyl phthalate (DEHP) (median 2.3 µg/kg bw/day; 95th percentile 8.9 µg/kg bw/day) in 5-year-old children. The tolerable daily intake (TDI) set by the European Food Safety Authority (EFSA) was exceeded in case of DnBP (in 1% of 9-year-olds and in 3% of 5-year-olds). Exposure risk was assessed based on hazard quotients calculation and cumulative approach for similar health effect. The combined exposure to four phthalates expressed by hazard index (HI) for reprotoxicity revealed exceeding of HI threshold in 14% of 5-year-olds and in 9% of 9-year-olds. These findings strongly support the need to reduce the burden of children by phthalates.