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. 2017 Oct;822:27-33.
doi: 10.1016/j.mrgentox.2017.07.003. Epub 2017 Jul 16.

Personal Samplers of Bioavailable Pesticides Integrated With a Hair Follicle Assay of DNA Damage to Assess Environmental Exposures and Their Associated Risks in Children

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Personal Samplers of Bioavailable Pesticides Integrated With a Hair Follicle Assay of DNA Damage to Assess Environmental Exposures and Their Associated Risks in Children

Pierre-Alexandre Vidi et al. Mutat Res. .
Free PMC article

Abstract

Agriculture in the United States employs youth ages ten and older in work environments with high pesticide levels. Younger children in rural areas may also be affected by indirect pesticide exposures. The long-term effects of pesticides on health and development are difficult to assess and poorly understood. Yet, epidemiologic studies suggest associations with cancer as well as cognitive deficits. We report a practical and cost-effective approach to assess environmental pesticide exposures and their biological consequences in children. Our approach combines silicone wristband personal samplers and DNA damage quantification from hair follicles, and was tested as part of a community-based participatory research (CBPR) project involving ten Latino children from farmworker households in North Carolina. Our study documents high acceptance among Latino children and their caregivers of these noninvasive sampling methods. The personal samplers detected organophosphates, organochlorines, and pyrethroids in the majority of the participants (70%, 90%, 80%, respectively). Pesticides were detected in all participant samplers, with an average of 6.2±2.4 detections/participant sampler. DNA damage in epithelial cells from the sheath and bulb of plucked hairs follicles was quantified by immunostaining 53BP1-labled DNA repair foci. This method is sensitive, as shown by dose response analyses to γ radiations where the lowest dose tested (0.1Gy) led to significant increased 53BP1 foci density. Immunolabeling of DNA repair foci has significant advantages over the comet assay in that specific regions of the follicles can be analyzed. In this cohort of child participants, significant association was found between the number of pesticide detections and DNA damage in the papilla region of the hairs. We anticipate that this monitoring approach of bioavailable pesticides and genotoxicity will enhance our knowledge of the biological effects of pesticides to guide education programs and safety policies.

Keywords: Children and adolescents; Environmental justice; Genotoxicity; Health outcomes predictions; Pesticides; Silicone wristbands.

Conflict of interest statement

Conflict of interest

Kim Anderson, an author of this research, discloses a financial interest in MyExposome, Inc., which is marketing products related to the research being reported. The terms of this arrangement have been reviewed and approved by Oregon State University in accordance with its policy on research conflicts of interest.

Figures

Fig. 1
Fig. 1
Detection of DNA breaks in plucked hair follicles. (A) Bright field images and nuclei staining (DAPI) in hairs from the eyebrows and the scalp. Arrowheads indicate the outer sheath region (S) and hair tip (T). (B) Confocal microscopy images of 53BP1 and γH2AX staining in the sheath (top) or tip (bottom) of hairs plucked from the scalp, and either irradiated (IR, 3 Gy) or left untreated (control). Single cell nuclei are shown in the insets. (C) Quantification of 53BP1 foci per nucleus cross-section as a function of IR doses. Different letters indicate significant differences (P < 0.01; Kruskal-Wallis and Dunn’s multiple comparison test, N > 100 nuclei from 5 different hairs). (D) Proliferation of hair follicular cells. Ki67 staining of a hair tip and sheath is shown. A mitotic figure is displayed in the inset. Scale bars, 200 μm (A, D [top]) or 20 μm (B and D [bottom]).
Fig. 2
Fig. 2
Association between pesticide exposures and DNA damage in children hair follicles. (A) Staining of a child participant hair with antibodies against 53BP1 (DNA damage) and NuMA (staining control). The arrowhead points to a nucleus with DNA repair foci. Nuclei were counterstained with DAPI. (B) Illustration of the sheath (S) and tip (T) regions of a scalp hair follicle plucked from a participant. (C) DNA damage (average number of 53BP1 foci/nucleus cross-section ± SEM) in the sheath and tip regions. (D) DNA damage in hair sheaths or at hair tips, plotted against the number of pesticides detected with PSD in each participant. (E) Confocal images of 53BP1 staining (left) and DNA damage quantification (right) in participants with or without detection of pesticides described as carcinogenic by Cal/EPA. Individual values are plotted and means are indicated. Scale bars, 20 μm.

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