Garrett A. Keating

U.S. dietary exposures to heterocyclic amines.

Heterocyclic amines (HAs) formed in fried, broiled or grilled meats are potent mutagens that increase rates of colon, mammary, prostate and other cancers in bioassay rodents. Studies of how human dietary HA exposures may affect cancer risks have so far relied on fairly crudely defined HA-exposure categories. Recently, an integrated, quantitative approach to HA-exposure assessment (HAEA) was developed to estimate compound-specific intakes for particular individuals based on corresponding HA-concentration estimates that reflect their meat-type, intake-rate, cooking-method and meat-doneness preferences. This method was applied in the present study to U.S. national Continuing Survey of Food Intakes by Individuals (CSFII) data on meats consumed and cooking methods used by >25,000 people, after adjusting for underreported energy intake and conditional on meat-doneness preferences estimated from additional survey data. The U.S. population average lifetime time-weighted average of total HAs consumed was estimated to be ∼9 ng/kg/day, with 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP) estimated to comprise about two thirds of this intake. Pan-fried meats were the largest source of HA in the diet and chicken the largest source of HAs among different meat types. Estimated total HA intakes by male vs. female children were generally similar, with those by (0- to 15-year-old) children ∼25% greater than those by (16+-year-old) adults. Race-, age- and sex-specific mean HA intakes were estimated to be greatest for African American males, who were estimated to consume ∼2- and ∼3-fold more PhIP than white males at ages <16 and 30+ years, respectively, after considering a relatively greater preference for more well-done items among African Americans based on national survey data. This difference in PhIP intakes may at least partly explain why prostate cancer (PC) kills ∼2-fold more African American than white men, in view of experimental data indicating that PhIP mutates prostate DNA and causes prostate tumors in rats.

Factors affecting human heterocyclic amine intake and the metabolism of PhIP

We are working to understand possible human health effects from exposure to heterocyclic amines that are formed in meat during cooking. Laboratory-cooked beef, pork, and chicken are capable of producing tens of nanograms of MeIQx, IFP, and PhIP per gram of meat and smaller amounts of other heteroyclic amines. Well-done restaurant-cooked beef, pork, and chicken may contain PhIP and IFP at concentrations as high as tens of nanograms per gram and MeIQx at levels up to 3 ng/g. Although well-done chicken breast prepared in the laboratory may contain large amounts of PhIP, a survey of flame-grilled meat samples cooked in private homes showed PhIP levels in beef steak and chicken breast are not significantly different (P=0.36). The extremely high PhIP levels reported in some studies of grilled chicken are not seen in home-cooked samples.Many studies suggest individuals may have varying susceptibility to carcinogens and that diet may influence metabolism, thus affecting cancer susceptibility. To understand the human metabolism of PhIP, we examined urinary metabolites of PhIP in volunteers following a single well-done meat exposure. Using solid-phase extraction and LC/MS/MS, we quantified four major PhIP metabolites in human urine. In addition to investigating individual variation, we examined the interaction of PhIP with a potentially chemopreventive food. In a preliminary study of the effect of broccoli on PhIP metabolism, we fed chicken to six volunteers before and after eating steamed broccoli daily for 3 days. Preliminary results suggest that broccoli, which contains isothiocyanates shown to induce Phases I and II metabolism in vitro, may affect both the rate of metabolite excretion and the metabolic products of a dietary carcinogen. This newly developed methodology will allow us to assess prevention strategies that reduce the possible risks associated with PhIP exposure.

Factors determining dietary intakes of heterocyclic amines in cooked foods.

The identification of heterocyclic amines (HCAs) in cooked foods has focused attention on the potential health effects from their consumption in the diet. Recent studies have estimated daily dietary intakes of HCAs that vary 10-fold and implicated different cooked meats as the prime source of HCAs in the diet. These varied estimates can be attributed to the different dietary assessment methods used in these studies, as well as the different levels of HCAs ascribed to the most commonly consumed cooked meats. Epidemiological studies utilizing information on dietary practice and food intake have found higher risks for several cancers among individuals consuming the highest levels of HCAs. These studies have highlighted the importance of using information on cooking methods in addition to food intake to accurately estimate dietary exposure to HCAs.

Measured and estimated air concentrations of chloroform in showers: Effects of water temperature and aerosols

Abstract Air concentrations of chloroform (CHCl3) in an experimental shower chamber were compared with estimates predicted by determining CHCl3 loss between the shower nozzle and drain and treating the shower chamber as a single, well-mixed compartment. CHCl3 water-to-air transfer efficiencies for experimental nozzles producing droplets of 1800 and 300 μm diameters were 0.90 and 0.80, respectively, and 0.65 for a standard domestic shower nozzle. Significant differences were observed in the transfer efficiencies when the experimental nozzles were operated at 35 and 45°C but not when the standard nozzle was operated in either jet or spray flow at 40°C. Aerosol concentrations were measured continuously with a hot-wire probe and ranged between 3000 and 17,000 aerosols/cm3 for the three nozzles. Measured air concentrations showed variable agreement with predicted values; under certain conditions, air concentrations were between 10 and 50% less than the predicted values. These discrepancies were observed at the highest aerosol concentrations and when peak aerosol concentrations occurred between 2 and 4 min of the 10 min shower.

Quantification of the Neurotoxic β-Carboline Harmane in Barbecued/Grilled Meat Samples and Correlation with Level of Doneness

Harmane, one of the heterocyclic amines (HCAs), is a potent neurotoxin linked to human diseases. Dietary exposure, especially in cooked meats, is the major source of exogenous exposure for humans. However, knowledge of harmane concentrations in cooked meat samples is limited. Our goals were to (1) quantify the concentration of harmane in different types of cooked meat samples, (2) compare its concentration to that of other more well-understood HCAs, and (3) examine the relationship between harmane concentration and level of doneness. Thirty barbecued/grilled meat samples (8 beef steak, 12 hamburger, 10 chicken) were analyzed for harmane and four other HCAs (2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine [PhIP], amino-3,8-dimethylimidazo[4,5-f]quinoxaline [MeIQx], 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline [DiMeIQx], and 2-amino-1,6-dimethylfuro[3,2-e]imidazo[4,5-b]pyridine [IFP]). Mean (± SD) harmane concentration was 5.63 (± 6.63) ng/g; harmane concentration was highest in chicken (8.48 ± 9.86 ng/g) and lowest in beef steak (3.80 ± 3.6 ng/g). Harmane concentration was higher than that of the other HCAs and significantly correlated with PhIP concentration. Harmane concentration was associated with meat doneness in samples of cooked beef steak and hamburger, although the correlation between meat doneness and concentration was greater for PhIP than for harmane. Evidence indicates that harmane was detectable in nanograms per gram quantities in cooked meat (especially chicken) and, moreover, was more abundant than other HCAs. There was some correlation between meat doneness and harmane concentration, although this correlation was less robust than that observed for PhIP. Data such as these may be used to improve estimation of human dietary exposure to this neurotoxin.

Dietary Epidemiology of Essential Tremor: Meat Consumption and Meat Cooking Practices

Background/Aim: Harmane [1-methyl-9H-pyrido(3,4-b)indole] is a tremor-producing neurotoxin. Blood harmane concentrations are elevated in essential tremor (ET) patients for unclear reasons. Potential mechanisms include increased dietary harmane intake (especially through well-cooked meat) or genetic-metabolic factors. We tested the hypothesis that meat consumption and level of meat doneness are higher in ET cases than in controls. Methods: Detailed data were collected using the Lawrence Livermore National Laboratory Meat Questionnaire. Results: Total current meat consumption was greater in men with than without ET (135.3 ± 71.1 vs. 110.6 ± 80.4 g/day, p = 0.03) but not in women with versus without ET (80.6 ± 50.0 vs. 79.3 ± 51.0 g/day, p = 0.76). In an adjusted logistic regression analysis in males, higher total current meat consumption was associated with ET (OR = 1.006, p = 0.04, i.e., with 10 additional g/day of meat, odds of ET increased by 6%). Male cases had higher odds of being in the highest than lowest quartile of total current meat consumption (adjusted OR = 21.36, p = 0.001). Meat doneness level was similar in cases and controls. Conclusion: This study provides evidence of a dietary difference between male ET cases and male controls. The etiological ramifications of these results warrant additional investigation.

Inhibiting the Transport of Hazardous Spores Using Polymer-Based Solutions

A series of polymer solutions were developed for the purpose of immobilizing aerosolized 1–10 μ m sized hazardous biological particles. The polymer solutions were designed as tools for emergency response and remediation personnel. The inhibition of secondary aerosolization and migration of biothreat particles has important implications for public health protection and contamination cleanup. Limiting further dispersion of particles such as Bacillus anthracis spores may reduce inhalation hazards and enhance remediation efficiencies. This study evaluated film-forming polymers that have multiple functional groups capable of attracting and binding particles; these included acrylates, cellulosics, vinyl polymers, and polyurethanes. The selected polymers were combined with appropriate solvents to design solutions that met specific performance objectives. The polymer solutions were then evaluated for key characteristics, such as high adhesion, high elasticity, low density, short drying time, low viscosity, and low surface tension. These solutions were also evaluated for their adhesion to biothreat agent in a series of wind tunnel experiments using highly refined aerosolized Bacillus atrophaeus spores (a simulant for anthrax, 1–3 μ m). Results demonstrated that a polymer solution, an amphoteric acrylate identified as NS-2, was the best candidate for attaching to spores and inhibiting reaerosolization. This polymer solution was anionic, thus providing the electrostatic (coulombic) attraction to cationic spores, had low surface tension, and performed well in wind tunnel tests.