Influence of fatty acid diets on gene expression in rat mammary epithelial cells.

BACKGROUND: This study examines the impact of dietary fatty acids on regulation of gene expression in mammary epithelial cells before and during puberty. METHODS: Diets primarily consisted of n-9 monounsaturated fatty acids (olive oil), n-6 polyunsaturated fatty acids (safflower), saturated acids (butter), and the reference AIN-93G diet (soy oil). The dietary regimen mimics the repetitive nature of fatty acid exposure in Western diets. Diet-induced changes in gene expression were examined in laser capture microdissected mammary ductal epithelial cells at day of weaning and end of puberty. PCNA immunohistochemistry analysis compared proliferation rates between diets. RESULTS: Genes differentially expressed between each test diets and the reference diet were significantly enriched by cell cycle genes. Some of these genes were involved in activation of the cell cycle pathway or the G2/M check point pathway. Although there were some differences in the level of differential expression, all diets showed qualitatively the same pattern of differential expression compared to the reference diet. Cluster analysis identified an expanded set of cell cycle as well as immunity and sterol metabolism related clusters of differentially expressed genes. CONCLUSION: Fatty acid-enriched diets significantly upregulated proliferation above normal physiological levels during puberty. Higher cellular proliferation during puberty caused by enriched fatty acid diets poses a potential increase risk of mammary cancer in later life. The human homologs of 27 of 62 cell cycle rat genes are included in a human breast cancer cluster of 45 cell cycle genes, further emphasizing the importance of our findings in the rat model.

Early exposure to lead and juvenile delinquency.

Cross-sectional studies have reported an association between lead (Pb) levels in bone and delinquent behavior in later childhood and adolescence. This is the first prospective longitudinal study of Pb and child development to address this question with comprehensive assessments of toxicant exposure and other developmental cofactors. A prospective longitudinal birth cohort of 195 urban, inner-city adolescents recruited between 1979 and 1985 was examined. Relationships between prenatal and postnatal exposure to Pb (serial blood Pb determinations) and antisocial and delinquent behaviors (self- and parental reports) were examined. Prenatal exposure to Pb was significantly associated with a covariate-adjusted increase in the frequency of parent-reported delinquent and antisocial behaviors, while prenatal and postnatal exposure to Pb was significantly associated with a covariate-adjusted increase in frequency of self-reported delinquent and antisocial behaviors, including marijuana use. Use of marijuana itself by Cincinnati Lead Study (CLS) teens was strongly associated with all measures of delinquent and antisocial behavior. This prospective longitudinal study confirmed earlier clinical observations and recent retrospective studies that have linked Pb exposure with antisocial behavior in children and adolescents. Both prenatal and postnatal exposure to Pb were associated with reported antisocial acts and may play a measurable role in the epigenesis of behavioral problems independent of the other social and biomedical cofactors assessed in this study.

A new anthropometric phantom for calibrating in vivo measurements of stable lead in the human leg using x-ray fluorescence.

A new anthropometric phantom has been developed for calibrating in vivo measurements of stable lead deposited in bone using x-ray fluorescence. The phantom reproduces the shape of the mid shaft of the adult human leg and is fabricated using polyurethanes and calcium carbonate to produce materials that exhibit the same density, energy transmission, and calcium content as cortical bone, bone marrow, and muscle. The phantom includes a removable tibia fabricated using simulants for cortical bone and bone marrow to which a precise amount of stable lead has been added to cortical bone. The formulations used in fabricating the new anthropometric phantom are much more uniform in density and composition than the conventional phantom made from Plexiglas cylinders filled with plaster-of-Paris. The energy spectrum from an x-ray fluorescence measurement of the phantom using a 109Cd source is indistinguishable from an in vivo x-ray fluorescence measurement of the human leg, demonstrating that the materials used in the phantom exhibit the same radiological properties as human tissue. Likewise, results from x-ray fluorescence measurements of the phantom exhibit the same positional dependency as the human leg and vary by approximately 36% when, for example, the phantom containing 54 ppm of stable lead in the tibia was rotated by only 15 degrees. The detection limit for a 30 min 109Cd K shell x-ray fluorescence in vivo measurement is approximately 20 ppm determined from a background measurement using the new phantom containing no added lead in the muscle, bone, or bone marrow. The new anthropometric phantom significantly improves in vivo x-ray fluorescence calibration measurements by (1) faithfully reproducing the anatomy of the human leg, (2) having components that exhibit radiological properties similar to that of human tissue, and (3) providing a realistic calibration standard that can be used for in vivo x-ray fluorescence intercomparison measurements.

Particle settling after lead-based paint abatement work and clearance waiting period.

This study investigated the evolution of airborne particle concentration and size distribution following abatement work in a controlled environment utilizing direct real-time particle monitoring and used it to project potential lead loadings as those particles settle. An 860 ft3 environmental test chamber with sophisticated ventilation and air purifying systems was built. Wooden doors with lead-based paint were dry sanded or scraped to generate the highest feasible airborne lead concentrations. Size-fractional airborne particle concentrations decreased exponentially with time in all tests, even with no air exchange, consistent with the stirred model of constantly mixed air, which predicts longer settling than for tranquil settling. Very low levels of air mixing generated by temperature gradients and initial room air turbulence affected particle settling. About 90% of airborne lead mass settled within 1 hour after active abatement, before final cleaning began. During the second waiting period of 1 hour, which followed cleaning of the floor, additional dust settled so that the additional potential lead loading from remaining airborne lead was less than 20 microg/ft2. For this worst case scenario, the underestimate of the lead loading done by the clearance sampling did not exceed about 30%. For more realistic conditions, the underestimates are projected to be much lower than the new 40 microg/ft2 Housing and Urban Development (HUD) clearance standards for floor dust lead. These results were obtained for the first waiting period (between the end of active abatement and the beginning of cleaning) of 1 hour, as recommended by HUD guidelines. Thus, this study demonstrates no need to increase either the first or second waiting period.

An empirical comparison of lead exposure pathway models.

Structural equation modeling is a statistical method for partitioning the variance in a set of interrelated multivariate outcomes into that which is due to direct, indirect, and covariate (exogenous) effects. Despite this model's flexibility to handle different experimental designs, postulation of a causal chain among the endogenous variables and the points of influence of the covariates is required. This has motivated the researchers at the University of Cincinnati Department of Environmental Health to be guided by a theoretical model for movement of lead from distal sources (exterior soil or dust and paint lead) to proximal sources (interior dust lead) and then finally to biologic outcomes (handwipe and blood lead). The question of whether a single structural equation model built from proximity arguments can be applied to diverse populations observed in different communities with varying lead amounts, sources, and bioavailabilities is addressed in this article. This reanalysis involved data from 1855 children less than 72 months of age enrolled in 11 studies performed over approximately 15 years. Data from children residing near former ore-processing sites were included in this reanalysis. A single model adequately fit the data from these 11 studies; however, the model needs to be flexible to include pathways that are not frequently observed. As expected, the more proximal sources of interior dust lead and handwipe lead were the most important predictors of blood lead; soil lead often had a number of indirect influences. A limited number of covariates were also isolated as usually affecting the endogenous lead variables. The blood lead levels surveyed at the ore-processing sites were comparable to and actually somewhat lower than those reported in the the Third National Health and Nutrition Examination Survey. Lessened bioavailability of the lead at certain of these sites is a probable reason for this finding.

The contribution of lead-contaminated house dust and residential soil to children's blood lead levels. A pooled analysis of 12 epidemiologic studies.

In 1992, the U.S. Congress passed the Residential Lead-Based Paint Hazard Reduction Act, which requires the promulgation of health-based dust lead and soil lead standards for residential dwellings to prevent undue lead exposure in children. Unfortunately, the levels of lead in house dust and soil that are associated with elevated blood lead levels among U.S. children remain poorly defined. This pooled analysis was done to estimate the contributions of lead-contaminated house dust and soil to children's blood lead levels. The results of this pooled analysis, the most comprehensive existing epidemiologic analysis of childhood lead exposure, confirm that lead-contaminated house dust is the major source of lead exposure for children. These analyses further demonstrate that a strong relationship between interior dust lead loading and children's blood lead levels persists at dust lead levels considerably below the U.S. Department of Housing and Urban Development's current postabatement standards and the Environmental Protection Agency's guidance levels. Finally, these analyses demonstrate that a child's age, race, mouthing behaviors, and study-site specific factors influence the predicted blood lead level at a given level of exposure. These data can be used to estimate the potential health impact of alternative health-based lead standards for residential sources of lead exposure.

Field testing of new aerosol sampling method with a porous curved surface as inlet.

A new aerosol sampling method, utilizing a porous curved surface as the sampling inlet, has recently been developed. Previous laboratory evaluations of this method have demonstrated its important features, such as low wind sensitivity and good filter collection uniformity. In this study a prototype incorporating the new method was evaluated in the field as a stationary and personal sampling device. The small sampler, utilizing a 25-mm filter is called the button aerosol sampler and was evaluated for collecting total airborne dust and fungal spores. The study was performed in nine poorly maintained inner-city houses during environmental cleanups at different cleanliness levels. The button sampler was used in parallel with the standard 37-mm closed-face filter cassette. Four collocated samplers of each type were tested at all sites as stationary samplers, and three samplers of each type were tested at two sites as personal samplers. Aerosol samples were collected on filters and analyzed using the gravimetric method for total dust and epifluorescence microscopy for fungal spores. The average particle concentration values measured with the button sampler and with the standard sampling cassette were found to correlate well within ranges of 10(1)-10(3) micrograms/m3 for total dust and 10(3)-10(5) spores/m3 for airborne fungi. The measurement results obtained with the new button sampler showed lower intersample variations of the measured concentration levels and higher uniformity of the particle deposits on the filters than those obtained with the standard cassette.

Urinary arsenic excretion as a biomarker of arsenic exposure in children.

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 microg/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.

Assessment of reproductive disorders and birth defects in communities near hazardous chemical sites. I. Birth defects and developmental disorders.

Members of the workgroup on birth defects and developmental disorders discussed methods to assess structural anomalies, genetic changes and mutations, fetal and infant mortality, functional deficits, and impaired fetal and neonatal growth. Tier 1 assessments for all five adverse reproductive outcomes consist of questionnaires and reviews of medical records rather than laboratory testing of biologic specimens. The work-group members noted a role for neurodevelopmental testing and for limited genetic studies, such as karyotyping in Tier 2 assessments. Emerging methodologies to identify chromosomal aberrations, DNA adducts, and repair inhibition were reserved for Tier 3.

Environmental arsenic exposure of children around a former copper smelter site.

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.

The relationship between surface dust lead loadings on carpets and the blood lead of young children.

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.

Effect of early lead exposure on children's postural balance.

This study investigated the effect of chronic exposure to lead on children's ability to maintain upright postural balance as a biological marker of lead-induced modifications of the neuromotor system. For this study, 162 six-year-old children, with a five-year geometric mean lead concentration in blood of 11.9 micrograms/dL (range 4.0-28.0 micrograms/dL), were tested for postural balance with a microprocessor-based force platform system. An increase in blood lead was significantly associated with an increase in the variable postural sway--implying poorer postural balance. This association was not influenced by socio-economic, racial or environmental factors. This simple, objective and quick technique may be useful for assessing gross motor functions in children who are at or below the United States Centers for Disease Control's class III category and/or for monitoring the effectiveness of medical interventions aimed at reversing lead-associated impairment of upright postural balance.

Clean-up of lead in household carpet and floor dust.

Methods to remove lead-containing dust were tested on carpets from homes of children with high blood lead and on new carpets artificially contaminated in the laboratory. The household carpets could not be cleaned effectively by repetitive vacuuming with HEPA-filtered cleaners. The lead concentration in the removed dust remained about the same from the initial cleaning (1 min/m2) to the final cleaning (total cleaning time of 10 min/m2). The lead loading on the surface of the carpets often increased during cleaning because vacuuming brought lead from deeper in the carpet to the surface. Over 95% of the total dust was removed from bare wooden floors by dry vacuuming (5 min/m2). For linoleum, more than 75% was removed by vacuuming for 5 min/m2. However, little was removed in vacuuming after the initial two minutes and about 20% was removed in a final wet-washing step. HEPA-vacuuming of the laboratory-contaminated carpets revealed that two of the commercially available vacuum cleaners tested were essentially equivalent and each removed significantly more dust than a third vacuum during a total cleaning time of 10 min/m2. Cleaning for 6 min/m2 was necessary to remove more than 70% of the embedded dust by the two more efficient vacuums. Cleaning efficiencies were about the same for short pile and sculptured carpets. It was concluded that it may be more practical to replace rather than clean carpets. HEPA-vacuum cleaning of carpets was shown to increase lead dust on the surface under some conditions.

Functional implications of postural disequilibrium due to lead exposure.

Measurement of postural equilibrium has been employed as an indirect indicator of functional status of the nervous systems of 109 children (mean age: 5.8 +/- 0.78 years) from the Cincinnati Lead Program Project. The geometric mean blood lead for the first five years of life (PbB05) was 11.9 +/- 1.5 micrograms/dL. Postural sway associated with upright balance was noninvasively quantitated with a microprocessor-based force platform and four tasks performed for 30 sec each by the subjects. A covariate-adjusted multiple regression analysis showed statistically significant associations between PbB05 and the postural sway for the task requiring primarily vestibular and/or proprioceptive systems input, implying potential functional impairment. These findings raised several new issues which are addressed here: (1) A dynamic task has been developed to further test the effect of perturbing those afferents' functional capabilities. (2) A method has been developed to quantitate the stability boundary of each subject to better characterize the limits of functionally-safe postural sway. (3) There is a need to perform the postural sway in a shorter duration than 30 sec so that children younger than five years of age can be tested for early identification of Pb-induced functional impairment of postural equilibrium.

The developmental consequences of low to moderate prenatal and postnatal lead exposure: intellectual attainment in the Cincinnati Lead Study Cohort following school entry.

In a further follow-up study of the Cincinnati Lead Study Cohort, 253 children were administered the Wechsler Intelligence Scale for Children-Revised (WISC-R) at approximately 6.5 years of age. Postnatal blood lead concentrations were inversely associated with Full-Scale (FSIQ) and Performance IQ (PIQ). Following statistical adjustment for developmental co-factors such as maternal IQ and an assessment of the quality of caretaking in the home environment, a statistically significant relationship remained between postnatal blood lead concentrations and PIQ. Further statistical analyses suggested that averaged lifetime blood lead concentrations in excess of 20 micrograms/dL were associated with deficits in PIQ on the order of approximately 7 points when compared to children with mean concentrations less or equal to 10 micrograms/dL. These results are discussed in terms of their consistency with other similar studies as well as their internal consistency with earlier reports on this cohort. The findings of this investigation support recent initiatives in the United States to reduce the exposure of children to environmental lead.

Role of exposure databases in epidemiology.

At present, exposure databases record data primarily for regulatory purposes; they have not focused on serving the needs of epidemiologists or public health. However, the modification of exposure databases could facilitate their use in epidemiology. Characteristics necessary to enhance the use of all databases include easy access by users; documentation of methods, sampling bias, error, and inconsistences; widespread coverage in time and space; and methods and measures for estimating exposure of individuals as well as populations. Also needed are exposure scenarios and models to estimate exposures for geographic areas and time intervals not currently sampled. Multidisciplinary teams are needed to examine current databases, to review strategies for improving data collection, and to suggest and help implement appropriate changes. A long-term goal is to develop and validate data from exposure scenarios and models using data on the relationship of exposure to doses measured in humans.

Lead exposure and growth in the early preschool child: a follow-up report from the Cincinnati Lead Study.

This report is a follow-up of an earlier study of the effects of low to moderate prenatal and postnatal lead exposure on children's growth in stature. Two hundred thirty-five subjects were assessed every 3 months for lead exposure (blood lead level) and stature (recumbent length) up to 33 months of age. Fetal lead exposure was indexed by maternal blood lead level during pregnancy. The adverse effects of lead on growth during the first year of life were reported previously. This analysis covers essentially the second and third years of life. The results indicate that mean blood lead level during this period was negatively associated with attained height at 33 months of age (P = .002). This association was, however, evidenced only among those children who had mean blood lead levels greater than the cohort median (greater than or equal to 10.77 micrograms/dL) during the 3- to 15-month interval. The results also suggest that the effect of lead exposure (both in utero as well as during the first year of life) are transient provided that subsequent exposure to lead is not excessive. It appears that maintaining an average blood lead level of 25 micrograms/dL or more during the second and third year of life was detrimental to the child's attained stature at 33 months of age. Approximately 15% of this cohort experienced these levels of lead exposure. Continued follow-up of this cohort will reveal whether these lead-related deficits persist and whether they continue to be dependent on the level of exposure in an earlier period.

Serum vitamin D metabolites and bone mineralization in young children with chronic low to moderate lead exposure.

One hundred five children (49 male, 99 black) with known lead exposure indices from birth and adequate nutrient intake of calcium, phosphorus, and vitamin D were studied at 1 of 3 ages (21, 27, or 33 months) to determine the effects of chronic low to moderate lead exposure on circulating concentrations of vitamin D metabolites and bone mineral content as determined by photon absorptiometry. Univariate multiple regression analyses showed no direct relationship of blood lead levels to vitamin D metabolites or bone mineral content. Structural equation analyses which took into account potential covariates of age, season, race, and sex showed estimated declines in serum concentrations of total calcium (from 9.72 to 9.61 mg/dL), phosphorus (from 5.4 to 4.67 mg/dL), and 25-hydroxyvitamin D (from 27.24 to 25.8 ng/mL) and estimated increases in concentrations of parathyroid hormones (from 73.03 to 83.14 microL Eq/mL), 1,25-dihydroxyvitamin D (from 62.39 to 62.69 pg/mL), and bone mineral content (from 222.66 to 234.91 mg/cm) over the observed range of average lifetime blood lead concentrations (4.76 to 23.61 micrograms/dL, geometric mean 9.74 micrograms/dL). However, the only statistically significant effect of average lifetime blood lead concentration was that for phosphorus, and the multivariate test of the combined effects of lead on these six outcomes was not statistically significant (P = .2). It is concluded that significant alterations in vitamin D metabolism, calcium and phosphorus homeostasis, and bone mineral content are not present in children whose nutritional status is adequate and who experience low to moderate lead exposure.

Lead exposure and the cognitive development of urban preschool children: the Cincinnati Lead Study cohort at age 4 years.

The purpose of this analysis was to determine if significant associations could be observed between prenatal/postnatal blood lead (PbB) levels and the cognitive development of 258 urban, inner-city children at 4 years of age. These children have been followed since birth with frequent assessments of general health, PbB, and neuropsychological status. The Kaufman Assessment Battery for Children (K-ABC) was administered at approximately 4 years of age. Higher neonatal PbB levels were associated with poorer performance on all K-ABC subscales. However, this inverse association was limited to children from the poorest families. Maternal PbB levels were unrelated to 4-year cognitive status. Few statistically significant associations between postnatal PbB levels and K-ABC scales could be found. However, the results did suggest a weak inverse relationship between postnatal PbB levels and performance on a K-ABC subscale which assesses visual-spatial and visual-motor integration skills. In these results we note both contradiction and accord with previously published prospective studies.

Lead exposure and neurobehavioral development in later infancy.

A prospective methodology was used to assess the neurobehavioral effects of fetal and postnatal lead exposure during the first 2 years of life. Lead was measured in whole blood prenatally in mothers and at quarterly intervals in the infant. Prenatal blood lead levels were low (mean = 8.0 micrograms/dL). However, approximately 25% of the study infants had at least one serial blood lead level of 25 micrograms/dL or higher during the second year of life. Multiple regression and structural equation analyses revealed statistically significant relationships between prenatal and neonatal blood lead level and 3- and 6-month Bayley Mental and/or Psychomotor Development Index. However, by 2 years of age, no statistically significant effects of prenatal or postnatal lead exposure on neurobehavioral development could be detected. Data consistent with the hypothesis that a postnatal neurobehavioral growth catch-up occurred in infants exposed fetally to higher levels of lead are presented.