Human studies suggest that high-fat diets (HFDs) increase the risk of breast cancer. The 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis rat model is commonly used to evaluate the effects of lifestyle factors such as HFD on mammary tumor risk. Past studies focused primarily on the effects of continuous maternal exposure on the risk of offspring at the end of puberty (PND50). We assessed the effects of prenatal HFD exposure on cancer susceptibility in prepubertal mammary glands and identified key gene networks associated with such disruption. During pregnancy, dams were fed AIN-93G-based diets with isocaloric high olive oil, butterfat or safflower oil. The control group received AIN-93G. Female offspring were treated with DMBA on PND21. However, a significant increase in tumor volume and a trend of shortened tumor latency were observed in rats with HFD exposure against the controls (P=.048 and P=.067, respectively). Large-volume tumors harbored carcinoma in situ. Transcriptome profiling identified 43 differentially expressed genes in the mammary glands of the HFBUTTER group as compared with control. Rapid hormone signaling was the most dysregulated pathway. The diet also induced aberrant expression of Dnmt3a, Mbd1 and Mbd3, consistent with potential epigenetic disruption. Collectively, these findings provide the first evidence supporting susceptibility of prepubertal mammary glands to DMBA-induced tumorigenesis that can be modulated by dietary fat that involves aberrant gene expression and likely epigenetic dysregulation.

PFC serum concentrations were measured in 6-8 year-old girls in Greater Cincinnati (GC) (N = 353) and the San Francisco Bay Area (SFBA) (N = 351). PFOA median concentration was lower in the SFBA than GC (5.8 vs. 7.3 ng/mL). In GC, 48/51 girls living in one area had PFOA concentrations above the NHANES 95th percentile for children 12-19 years (8.4 ng/mL), median 22.0 ng/mL. The duration of being breast fed was associated with higher serum PFOA at both sites and with higher PFOS, PFHxS and Me-PFOSA-AcOH concentrations in GC. Correlations of the PFC analytes with each other suggest that a source upriver from GC may have contributed to exposures through drinking water, and water treatment with granular activated carbon filtration resulted in less exposure for SWO girls compared to those in NKY. PFOA has been characterized as a drinking water contaminant, and water treatment systems effective in removing PFCs will reduce body burdens.

BACKGROUND: Polyfluoroalkyl compounds (PFCs) have been detected in human breast milk which is a major exposure source for lactating infants. The objective of this study was to determine if being breast fed was related to PFC serum concentrations in young girls. METHODS: Through the NIH Breast Cancer and the Environment Research Program (BCERP), we conducted a study of PFC biomarkers in serum of 6-8 year-old girls in Greater Cincinnati (GC) (N=353) and the San Francisco Bay Area (SFBA) (N=351), collected in 2005-2009. Mothers self-reported the duration of breast feeding their daughters. RESULTS: We detected five PFCs in >95% of the girls. Median serum concentrations of both perfluorohexanesulfonate (PFHxS) and perfluorooctanoate (PFOA) were higher in the GC girls than in the SFBA girls (PFHxS 5.2 vs. 2.3 ng/ml, p<0.001; PFOA 7.3 vs. 5.8 ng/ml, p<0.001) but were similar for other PFCs. In linear regression analyses, with race, education, body mass index, parity and age at sample date as covariates, being breast fed was associated positively with PFC serum concentration in both GC and the SFBA (p<0.01). Duration of breastfeeding also was significantly associated with higher PFOA concentration in girls from both sites, with a significant test for linear trend of ? estimates over the increasing duration categories (1-3, 4-6, 7-9, 10-12 and 13+ months) (SGBA p=0.03; GC p<0.0001). In GC, all breast feeding duration groups had p values ?0.03, and the longest duration category of 13+ months having a p of <0.0001. In analyses of the other PFCs with the same terms in the models, we found evidence for an association of breast feeding with higher serum concentrations of PFHxS, 2-(N-methyl-perfluorooctane sulfonamide) acetate, and perfluorooctane sulfonate in GC girls only. CONCLUSIONS: Exposure to PFCs through breast milk is associated with higher serum concentrations in childhood.

Background: Polyfluroalkyl compounds (PFCs) are bioaccumulative; health effects may include impaired liver function, cholesterol metabolism, pubertal development and cancer. An industrial plant in Parkersburg, WV discharged PFCs into the Ohio River beginning in 1980; elevated PFCs were detected in Ohio River water samples in 2009. Communities immediately downstream of the Parkersburg plant and a cohort of girls in Cincinnati, OH have elevated serum PFOA. Aims: Determine serum PFC concentrations in the Ohio River Valley population and identify potential exposure modifiers. Methods: Participants were recruited around Huntington, WV, Portsmouth, OH and Cincinnati, OH from May 2009 to January 2012, and provided water consumption, health effects, and residential history data. Water sources were identified through records searches and meetings with utilities. Serum samples were analyzed for 11 PFCs. Associations between serum PFC and recruitment site, sex, age, sample collection time, drinking water source, water consumption and water filter use were analyzed using ANCOVA. Results: Participants (N=370) were 2-85 years old, 72% female and 96% Caucasian, and used the Ohio River/Ohio River Aquifer as a drinking water source from 0 to 30+ years. 96% of participants drank tap water; 87% drank some bottled water. PFHxS (median=1.4ng/mL), PFNA (median=1.2ng/mL), PFOA (median=8.1ng/mL) and PFOS (median=7.1ng/mL) were detected in >99% of participants; PFDeA and Me-PFOSA-AcOH (medians=0.2ng/mL) were detected in >50%. All site median values for PFOA serum concentration (Huntington=7.8ng/mL; Portsmouth=8.8ng/mL; Cincinnati=6.1ng/ml) were above the 2009-2010 NHANES 90th percentile. Preliminary analysis found a significant association between serum PFOA level and years of using the Ohio River as a water source. Conclusions: Ohio River Valley residents have elevated PFOA exposure, but not other PFCs, in line with other studies. These findings suggest a major component of PFOA exposure is drinking water from the Ohio River.

Background and Aims: Polyfluoroalkyl compounds (PFCs), such as perfluorooctanate (PFOA) and perfluorooctane sulfonate (PFOS), have been reported to change mammary gland structure in laboratory animals and pubertal timing in humans. We explored the relationships between serum PFOA and PFOS concentrations and pubertal maturation in young girls. Methods: Within the NIH Breast Cancer and the Environment Research Program, we conducted a study of PFC environmental biomarkers, including PFOA and PFOS, in young girls (age 6-7 years at entry) from two sites (N=688 girls). Pubertal staging (breast and pubic hair) has been conducted by clinicians or trained research staff, every year or more frequently, for as long as six years. Using longitudinal analysis methods (time to event) we studied the relationship between PFOA and PFOS serum concentrations measured at the beginning of the study (2005-2006) with age at pubertal Stage 2. Results: Median serum concentrations of PFOA and PFOS differed at the two sites (California: PFOA= 5.8 ng/ml, PFOS=12.5 ng/ml vs. Cincinnati PFOA =7.9 ng/ml, PFOS 14.0 ng/ml). Among girls at the Cincinnati site, we found a statistically significant relationship between PFOA serum concentration and age at Tanner breast Stage 2, and an inverse relationship between PFOS and age when Tanner pubic hair Stage 2 was first noted, with adjustment for race/ethnicity, body mass index percentile for age, caregiver education and mother’s age at menarche. Neither of these relationships reached statistical significance among the girls at the California site. Conclusions: PFOA and PFOS may act as endocrine disruptors, affecting the timing of pubertal maturation in humans. Support for this project provided by the National Institute of Environmental Health Sciences and the National Cancer Institute, to the University of Cincinnati/Cincinnati Children’s Hospital Medical Center, Breast Cancer & the Environment Research Center (U01 ES12770), and Center for Environmental Genetics (P30-ES06096).

Background and Aims: Perfluorooctanoic acid (PFOA) is used in the production of stain and grease-resistant coatings; the human health effects of PFOA exposure are not well understood. An industrial plant in Parkersburg, WV discharged PFOA and other polyfluroalkyl compounds (PFCs) into the Ohio River from 1980 to 2004. Residents of communities near Parkersburg and girls living in the Greater Cincinnati area have serum PFOA concentrations higher than general population levels. This study examined PFOA exposure in communities along the Ohio River between Parkersburg and Cincinnati. Methods: Participants recruited from Huntington, WV, Portsmouth, OH and Fort Thomas, KY in 2009 and 2010 provided 15-mL blood samples to measure PFC concentrations and clinical parameters and answered questions about water consumption, health effects, and residential history. Drinking water sources were identified for each community through records searches and meetings with utilities. Associations between serum PFOA concentration and recruitment site, cumulative years drinking Ohio River water, self-reported daily tap water consumption, age and gender were analyzed using ANCOVA. Results: Serum has been analyzed for 193 participants from Huntington, WV (N=74), Portsmouth, OH (N=90), and Fort Thomas, KY (N=29); 61 additional samples are currently being analyzed. PFOA concentrations range from 0.4ng/mL to 62.7ng/mL (median 9.7ng/mL). Median serum PFOA concentrations for each site were greater than the 2007-2008 NHANES median (4.30 ng/mL). Preliminary analysis indicated a significant correlation between serum PFOA concentration and the cumulative number of years the participant used Ohio River water; residents with two or more years consuming Ohio River water had higher serum PFOA concentrations than residents who used other water sources. Conclusions: These findings suggest that communities along the Ohio River between Parkersburg and Cincinnati are exposed to PFOA and the major source is drinking water drawn from the Ohio River.

BACKGROUND:The Breast Cancer and the Environment Research Centers (BCERCs) include collaborators from basic sciences, epidemiology, and the community, conducting studies to investigate whether environmental exposures are associated with the timing of puberty. A pilot study of a subset of the study participants assessed the feasibility of measuring selected biomarkers of exposure in blood and urine in girls 6-8 years of age. In the Greater Cincinnati study population, we found an elevated serum concentration of perfluorooctanoate (PFOA) among > 90% of young girls living in a small community. OBJECTIVES:The research team deliberated whether and how to report the PFOA findings to our study families. We will address the issues considered in our decision, as well as the formats we used to present the findings. METHODS:The results were verified as we searched for potential sources of the elevated PFOA levels. As a research team, we grappled with issues regarding the reporting of unexpected results, derived from unknown sources and with unknown clinical significance. Ultimately, we did decide to present these findings to the study families through a well-developed communication plan. DISCUSSION:Research team members came from a variety of experiences and backgrounds, which led to different interpretations about the clinical, ethical, and public health issues surrounding these findings. The ethical debates centered around the precautionary principle, the right to know, and do no harm. CONCLUSIONS:Given advances in environmental biomarker technologies and greater use of the transdisciplinary research model, a communication plan must be developed for those involved as study participants.

The Evaluation of the US Department of Housing and Urban Development Lead-Based Paint Hazard Control Grant Program studied the effectiveness of the housing intervention performed in reducing the blood lead of children at four post-intervention times (6-months, 1-year, 2-years, and 3-years). A repeat measures analysis showed that blood lead levels declined up to three-years post-intervention. The results at each successive collection time were significantly lower than at the previous post-intervention time except for the difference between the levels at two and three years. At two-years post-intervention, geometric mean blood lead levels were approximately 37% lower than at pre-intervention. Children with pre-intervention blood lead levels as low as 10 μg/dL experienced substantial declines in blood lead levels. Previous studies have found substantial improvements only if a child's pre-intervention blood lead level was above 20 μg/dL. Individual interior lead hazard control treatments as grouped by Interior Strategy were not a significant predictor of post-intervention blood lead levels. However, children living in dwellings where exterior lead hazard control interventions were done had lower blood lead levels at one-year post-intervention than those living in dwellings without the exterior interventions (all other factors being equal), but those differences were only significant when the mean exterior paint lead loading at pre-intervention was about the 90th percentile (7.0mg/cm(2)). This observation suggests that exterior lead hazard control can be an important component of a lead hazard control plan. Children who were six to eleven months of age at pre-intervention had a significant increase in blood lead at one-year post-intervention, probably due to other exposures.

ISEE-0763 Background: Polyfluoroalkyl compounds (PFCs) and their salts, such as perfluorooctanoic acid (PFOA), have been reported to change mammary gland structure and function in laboratory animals. We explored the relationship between serum PFOA concentration and timing of pubertal maturation in young girls. Methods: Within the NIH Breast Cancer and the Environment Research Centers (BCERC), we conducted a study of multiple environmental biomarkers, including PFOA and other PFCs in serum of young girls (age 6–7 years at entry) from two sites (N = 689 girls). Pubertal staging (breast (B) and pubic hair (PH)) has been conducted by clinicians or trained research staff, every year or more frequently, for as long as four years. After calculating adjusted geometric means for all PFCs, we examined the relationship between PFOA serum concentration at the beginning of the study with body mass index (BMI) and pubertal Stage 2 at baseline and one year follow-up. Results: Detectable serum levels of five PFCs, including PFOA, were found in >95% of the girls. The PFOA median was 6.4 ng/ml (range < LOD 0.1 to 55.9 ng/ml), with 24.9% having values above the 95th percentile for children 12–19 years (NHANES 2003–2004 population (8.6 ng/ml)). At the follow-up visit, 28.3% of girls had reached Tanner stage B2+, 19.2% were PH2+ and 30.3% had a BMI percentile for age >85. In analyses where serum PFOA was modeled as a continuous variable, we found a direct relationship with pubertal breast status and an inverse relationship with BMI percentile at the follow-up visit, with adjustment for age, race, site and caregiver education. Conclusions: It appears that PFOA acts as an endocrine disruptor although perhaps not by the usual mechanism. Although the relationship with BMI was inverse, there was a direct relationship with breast maturation. We continue to explore these complex relationships in models including other covariates.

Measured trajectories of the Center Of Pressure (COP) on the feet have been used to study postural balance problems. Two new signal analysis methods, the Wavelet based Multiple Fractal Formalism (WMFF) and frequency spectrum envelop characterisation method, are developed to characterise the COP data in a set of two parameters with unique physical significances. The methods are applied to re-dissect the data obtained during the Cincinnati Lead Study (CLS) to gain additional insights into the effect of early lead-exposure on children's postural balance. It is found that the frequency spectrum envelope characterisation method is especially attractive and easy to implement.

This study investigated the influence of succimer chelation therapy in eliminating and/or minimizing lead-associated impairments of motor functions such as postural balance and locomotion or gait activities. In this study, postural balance and functional locomotion or gait were quantitated in 161 children in Cincinnati enrolled in a randomized, placebo-controlled, double blind clinical trial. In comparison to the placebo group, the succimer therapy group showed significantly decreased postural sway during dynamic task performance implying improved postural balance. The results from locomotion tests demonstrated significant improvements in functional tasks of obstacle crossing and normal walking in the succimer treated group. While some beneficial neuromotor effects of succimer therapy were observed in the present study there remains several unanswered questions such as how long these effects will persist and how succimer therapy modifies lead-associated cerebellar deficits manifesting as perturbations in vestibular and/or proprioception systems for postural balance and functional locomotion.

This prospective study investigated the impact of early exposure to lead on the maturation of children's postural balance. The effect of lead exposure on age-associated maturation of postural balance was investigated on 91 children from the Cincinnati Lead Study (CLS) with a 5-year geometric mean lead concentration in blood of 11.66 microg/dL (range 3.89-28.33 microg/dL) by re-assessing their postural balance approximately every 20 months starting at mean age of 6.6 years through mean age of 12.1 years. The results presented in this paper provide evidence that low to moderate lead exposure in early childhood has a measurable and statistically significant impact on the maturation of postural balance. In comparison to less exposed children, of those in the higher lead group showed an impaired postural balance response. The results from this study suggest that children with early childhood lead exposure may need additional time to approach (or "catch up" with) their maturational postural balance status. As these subjects are now adults in their early to mid-twenties, poor postural balance may impact their daily living tasks and pose a higher risk of potential injuries at home and work.

Arsenic residues in the communities surrounding former smelters remain a public health concern, especially for infants and children. To evaluate environmental exposure among these children, a population-based cross-sectional study was conducted in the vicinity of a former copper smelter in Anaconda, Montana. A total of 414 children less than 72 months old were recruited. First morning voided urine samples and environmental samples were collected for arsenic measurements. The geometric mean of speciated urinary arsenic was 8.6 microg/liter (GSD = 1.7, N = 289). Average arsenic levels of different types of soil ranged from 121 to 236 microg/g and were significantly related to proximity and wind direction to the smelter site. The same significant relationship was observed for interior dust arsenic. Speciated urinary arsenic was found to be significantly related to soil arsenic in bare areas in residential yards (P < 0.0005). In general, elevated excretion of arsenic was demonstrable and warranted parents' attention to reduce exposure of their children to environmental arsenic.

Urinary arsenic concentration has been used generally for the determination of exposure, but much concern has been raised over the most appropriate expression for urinary arsenic levels. In this study, we examined the influence of various adjustments of expressing urinary arsenic data. All children who were less than 72 mo of age and who were potty trained were invited to participate in the present study. Urine, soil, and dust samples Were collected, and arsenic measurements were made. The geometric mean of speciated urinary arsenic among children who provided first-voided urine samples on 2 consecutive mornings was 8.6 mu g/l (geometric standard deviation = 1.7, n = 289). Speciated urinary arsenic was related significantly to soil arsenic in bare areas (p <.0005). Use of a single urine sample versus the average of two first-voided urine samples collected on 2 consecutive mornings did not significantly alter the relationship between environmental arsenic and urinary arsenic levels. Furthermore, none of the adjustments to urinary concentration improved the strength of correlation between urinary arsenic and soil arsenic levels. Concentration adjustments may not be necessary for urinary arsenic levels obtained from young children who provide first-void samples in the morning.

The final clean-up of residential lead abatement projects in federally-supported housing, as well as in other housing in a number of states, must meet surface dust lead clearance levels expressed as μg of lead per square foot. These clearance levels were established because hand-to-mouth ingestion of lead-contaminated dust is recognised as a major pathway through which many children are exposed. A dilemma exists because many floors in housing undergoing abatement are carpeted and the established clearance levels are generally not recommended for use on carpets. These clearance levels are also used as 'action levels' to determine whether exposure reduction activities are needed. The US Environmental Protection Agency is currently in the process of issuing standards for hazardous levels of lead in interior dust and bare soil under Title X of the Housing and Community Development Act of 1992, ‘The Residential Lead-Based Paint Hazard Reduction Act of 1992’. An effort to develop a potential surface dust lead clearance level for carpets was made using an existing vacuum dust collection method that has previously been shown to be a reliable indicator of childhood lead exposure. This method was designed for use on carpeted and non-carpeted surfaces. Using data from the Cincinnati Soil Lead Abatement Demonstration Project, the suggested floor-dust lead level where an estimated 95% of the population of children would be expected to have blood lead values below the national goal of 10 μg dL−1, was more than an order of magnitude lower than the current floor-dust lead clearance level of 1080 μg m−2 (100 μg ft−2). Further comparisons of blood lead and carpet lead levels in other parts of the country should be performed before a risk-based lead loading clearance level is established.

The U.S. Department of Housing and Urban Development (HUD) and several states have established floor and window dust lead clearance levels that must be met following lead-based paint abatement. These levels are also used as action levels to determine when exposure reduction measures are needed. Data are lacking on the relationship between these levels and the Centers for Disease Control (CDC) goal of having no more than 5 percent of children with blood lead levels above 10 μg/dl. Similarly, little information is available on the relationship between results using the HUD-prescribed surface wipe dust sampling method and a vacuum dust collection method utilizing a personal air sampling pump as a vacuum source, which has been used in a number of lead exposure studies. Blood lead, paint lead, and floor dust lead levels by both methods from 53 households in a mining community with lead paint sources were examined to help answer these questions. Results suggest that the HUD floor clearance level may not be low enough to achieve the CDC childhood blood lead goal. The two dust collection methods were found to be statistically correlated. Vacuum dust lead was correlated to paint lead, but wipe dust lead was not.