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© 2006 American Society for Nutrition J. Nutr. 136:3054-3061, December 2006


Nutritional Epidemiology

Carotenoid and Tocopherol Estimates from the NCI Diet History Questionnaire Are Valid Compared with Multiple Recalls and Serum Biomarkers1

L. Beth Dixon2,*, Amy F. Subar3, Louise Wideroff3, Frances E. Thompson3, Lisa L. Kahle4 and Nancy Potischman3

2 Department of Nutrition, Food Studies, and Public Health, New York University, New York City, NY; 3 Division of Cancer Prevention and Population Sciences, National Cancer Institute, NIH, Bethesda, MD; and 4 Information Management Services, Silver Spring, MD

* To whom correspondence should be addressed. E-mail: beth.dixon{at}nyu.edu.


    ABSTRACT
 TOP
 ABSTRACT
 Introduction
 Methods
 Results
 Discussion
 LITERATURE CITED
 
To improve the measurement of usual dietary intake, the National Cancer Institute developed a cognitively based Diet History Questionnaire (DHQ), which has been validated against four 24-h dietary recalls (4 24-HR) for energy, macronutrients, and several vitamins and minerals. This analysis used data from The Eating at America's Table Study (EATS) to determine the validity of estimates for carotenoids and tocopherols from the DHQ. Over the course of a year, 163 participants provided 1 or 2 blood samples and completed the DHQ and 4 24-HR. For both the DHQ and the 4 24-HR, crude correlations between serum and diet were modest to strong for the provitamin A carotenoids ({alpha}-carotene, ß-carotene, ß-cryptoxanthin), low to modest for lycopene, and very low for lutein. The individual dietary tocopherols were weakly correlated with the serum tocopherols, but vitamin E from food and dietary supplements was strongly and positively correlated with serum {alpha}-tocopherol and strongly and inversely correlated with serum {gamma}-tocopherol for both instruments. Adjustment for energy, BMI, smoking status, serum total cholesterol, and serum triacylglycerol did not appreciably change the correlations. Using the method of triads, validity coefficients for the DHQ were comparable to the 4 24-HR and were especially strong for {alpha}-carotene, ß-cryptoxanthin, lutein + zeaxanthin, and total vitamin E in men and {gamma}-tocopherol and total vitamin E in women. In this study, there was no advantage of 2 blood samples over 1, suggesting reasonably stable ranking of individuals for these biomarkers, which is important for large epidemiologic studies that typically obtain only 1 blood sample for biomarker status.



    Introduction
 TOP
 ABSTRACT
 Introduction
 Methods
 Results
 Discussion
 LITERATURE CITED
 
In most large epidemiologic studies of diet and chronic disease, a food frequency questionnaire (FFQ)5 is used as an economical and practical method for collecting dietary intake data (1). The FFQ asks respondents to report their usual frequency of consumption of each item from a food list for a specified period of time. This information may be relatively accurate in terms of ranking individuals within the study, but the absolute intake may not be valid due to various types of errors, including biased reporting, entering a limited number of foods, missing culturally specific foods, and inaccuracies in nutrient databases. Validation or calibration studies of FFQ dietary intake estimates usually use another measure of dietary intake, such as multiple recalls or records as the reference instrument. Serologic markers of diet provide an alternative standard to which dietary questionnaires can be compared and provide the advantage of not requiring recall by the subjects. Nutritional biomarkers reflect habitual intake or a steady state of intake and metabolism (2).

In an effort to improve the measure of usual dietary intake, investigators at the National Cancer Institute (NCI) developed the Diet History Questionnaire (DHQ) using cognitive-based methods described previously (36). The Eating at America's Table Study (EATS) was designed to compare nutrient intakes estimated from the DHQ with 4 24-HR collected over the course of 1 y and with serum biomarkers collected 1–2 times during that year (7). Previous analyses of EATS data showed that the NCI DHQ produced correlations for energy, macronutrients, and several vitamins and minerals, consistent with other widely used FFQs (7). In this analysis, we compared estimates of carotenoids and tocopherols from the DHQ and from 4 24-HR with serum carotenoid and tocopherol concentrations in EATS participants who provided at least 1 fasting blood sample. We also used Kaak's method of triads (8) to determine validity coefficients for each serum–diet correlation of carotenoid and tocopherol.


    Methods
 TOP
 ABSTRACT
 Introduction
 Methods
 Results
 Discussion
 LITERATURE CITED
 
    Sample and study design. In August 1997, nationally representative sampling and random-digit dialing techniques were used to obtain 12,615 telephone numbers. Nonworking, nonresidential, unanswered numbers, and participants deemed ineligible (i.e., not aged 20–70 y or on liquid diets) were excluded. Of the 3590 eligible persons identified, 1640 completed a baseline questionnaire. Between September 1997 and August 1998, 4 24-HR were administered, timed to occur through 1 calendar year with 1 recall per season. Based on the date of enrollment, blood was drawn from a subsample of participants twice, between mo 4–5 and mo 10–11. The subsample was recruited in periodic waves, leading to a set of blood samples collected across the course of the year. After the year in which the 4 24-HR and blood samples were collected, all participants completed the DHQ. This study was approved by the NCI Special Studies Internal Review Board.

    Dietary measures. The 4 24-HR were collected using the multiple-pass methodology developed for the 1994–1996 Continuing Survey of Intakes by Individuals (CSFII) (9). Trained interviewers collected and coded the data using the University of Texas Food Intake Analysis System (FIAS), version 3.0. The majority of participants used standard measuring guides (e.g., measuring cups and spoons, ruler, 2-dimensional pictures) to report intakes.

The DHQ asked about frequency of intake over the past year for 124 food items and included portion sizes (10). Forty-four of the foods had embedded questions about seasonal intake, food type (e.g., low fat, diet, caffeine free), and fat uses or additions. The DHQ also included questions about the intake of low-fat foods and dietary supplements.

The 4 24-HR and the DHQ provided estimates for 6 individual dietary carotenoids ({alpha}-carotene, ß-carotene, ß-cryptoxanthin, lutein + zeaxanthin, and lycopene) and 2 individual dietary tocopherols ({alpha}-tocopherol and {gamma}-tocopherol) from food. The 4 24-HR and the DHQ also provided estimates for total ß-carotene equivalents, which is a summary measure that includes the provitamin A carotenoids: {alpha}-carotene, ß-carotene, and ß-cryptoxanthin from food and ß-carotene from dietary supplements. We derived an estimate for total vitamin E intake ({alpha}-tocopherol from food and dietary supplements) for these 2 dietary instruments.

Carotenoid and tocopherol estimates for foods reported on the 4 24-HR and the DHQ were determined from similar food items in the University of Minnesota Nutrition Data System for Research (NDSR) (11) using methodology developed by Dixon et al. (12). ß-carotene and vitamin E from dietary supplements were determined from the sum of single-nutrient supplements and from multivitamin use. For single-nutrient supplements for which dose was not reported and for multivitamins, nutrient values were assigned that corresponded to defaults recommended in NHANES 1999–2002: 5000 µg ß-carotene for single supplement, 180 mg {alpha}-tocopherol for vitamin E single supplement, 13.5 mg {alpha}-tocopherol of vitamin E and 0 µg ß-carotene for one-a-day types without minerals, and 13.5 mg {alpha}-tocopherol of vitamin E and 610 µg ß-carotene for one-a-day types with minerals. The mean dose of multivitamins per day was calculated by dividing the dose by the frequency of intake.

    Serum measures. A subsample of 240 participants was targeted for the blood collection in 4 U.S. metropolitan areas (Albany, New York; Tucson, Arizona; Indianapolis, Indiana; and Austin, Texas) participating from the larger study. Participants who provided blood specimens received a desktop calculator and $20 remuneration as incentives. Although 240 participants agreed to provide a blood sample, a total of 163 participants completed at least 1 blood draw (n = 155 at 4–5 mo, n = 144 at 10–11 mo, and n = 135 at both time points). Reasons for nonparticipation included refusal (n = 62), lost to follow-up for the main study (n = 14), or moved away (n = 1).

Participants were instructed to fast from midnight until the morning of the blood draw. Fasting status was confirmed by questionnaire at the time of the draw. Blood samples were allowed to clot at room temperature for 45 min and then centrifuged (2900 x g; 20 min), and serum was stored at –20°C or at –70°C until shipment to the main repository for long-term storage. All assays were conducted by the Medical Research Laboratories of Highland Heights, Kentucky. Serum carotenoids and tocopherols were measured by HPLC using the method of Kaplan et al. (13). Samples were run against standard references from the National Institute of Standards and Technology for quality control; values >2 SD were repeated. Serum cholesterol and triacylglycerol were measured by enzyme analysis using standard methods of the Lipid Research Clinics Program (14). Duplicate samples of in-house sera were run for quality control and unusual values of serum lipids were flagged and rechecked.

    Other measures. Data from the baseline questionnaire were used to determine age, race or ethnicity, highest level of education completed, smoking status (never, past, current), and self-reported BMI (kg/m2) of the participants.

    Statistical analysis. All analyses were stratified by sex. Frequencies of demographic and behavioral characteristics were calculated to describe the study sample. Serum and dietary carotenoids and tocopherols were log-transformed prior to calculating geometric means and 95% CI. Paired t tests were run to determine differences between serum values for Draw 1 and Draw 2. Spearman rank correlation coefficients were calculated between the serum nutrients at the first and second draws to determine the reproducibility of the serum data. Spearman rank correlation coefficients were also calculated between the respective serum and dietary nutrients and adjusted for energy, BMI, smoking status (never, former, current), serum total cholesterol, and serum triacylglycerol. Because underreporting of food intake may affect the association between serum and dietary nutrients, participants who likely underreported their energy intake were determined according to age- and sex-specific formulas derived for adults (15). Underreporters had a ratio of energy intake estimated from the mean of 4 24-HR to basal metabolic rate (BMRest) that was <1.06, a cut-point that is appropriate for 4 dietary recalls (16). Analyses of correlations were compared by strata of various factors including energy intake (adequate vs. underreporting), BMI (<25 kg/m2 or normal weight vs. BMI ≥25 kg/m2 or overweight) and smoking status (never vs. former or current smoker).

Validity coefficients between true intakes of carotenoids and tocopherols and estimated intakes from the DHQ, the 4 24-HR, and serum biomarkers were estimated using the method of triads (8). This method assumes that each of the 3 nutrient measures is linearly related to a latent factor corresponding to the true intake of the nutrient. Each validity coefficient is calculated from the correlations between each pair of methods and represents the square root of the proportion of variation in each nutrient measure that is related to true intake and also to the variation in the other 2 nutrient measures. Although the method of triads assumes that measurement errors from each instrument are mutually independent, random errors associated with a FFQ and 4 24-HR are likely related. Therefore, the correlations between nutrients estimated from both dietary instruments will be overestimated, and the validity coefficients are considered to be the upper limits of the true validity coefficients, whereas the correlations between nutrients from each dietary instrument and serum are considered to be the lower limits of the validity coefficients. For this study, we followed this reasoning to calculate the range for each validity coefficient.

To maximize sample size, the analytic sample in this study included 130 EATS participants (86 women, 44 men) who completed the DHQ and the 4 24-HR and had blood drawn at the first time period. For comparison, all results were repeated for participants who had their blood drawn at the second time period (82 women, 44 men) or at both time periods (77 women, 42 men). Analyses were conducted using SAS, version 8.2 (SAS Institute). For all analyses, {alpha} < 0.05 was used to determine significance.


    Results
 TOP
 ABSTRACT
 Introduction
 Methods
 Results
 Discussion
 LITERATURE CITED
 
The majority of EATS participants were Caucasian women between the ages of 40 and 59 y who completed >12 y of education, did not smoke, and were normal or overweight (Table 1). About one-third of EATS participants reported taking a daily multivitamin. About 24% of women and 14% of men took a daily vitamin E supplement. Few participants reported ever taking a single ß-carotene supplement.


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TABLE 1 Demographic and behavioral characteristics of women and men who had blood drawn at Draw 1 and who completed the DHQ and 4 24-HR in the Eating at America's Table Study, 1997–1998

 
All serum data were moderately to highly reproducible, with Spearman rank correlation coefficients of 0.68–0.84 between the first and second blood draws (Table 2). Geometric means of serum variables were similar between Draw 1 and Draw 2. Only serum ß-carotene and serum lutein + zeaxanthin in women with both draws differed between Draw 1 and Draw 2 (P < 0.05). The mean values from our analytic sample were also comparable to mean serum carotenoid and tocopherol concentrations from participants who had blood drawn at either or both time periods, but who did not complete the DHQ or all 4 24-HR (data not presented).


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TABLE 2 Geometric means and 95% CI for serum carotenoids, tocopherols, and lipids from women and men who had blood drawn at Draw 1, Draw 2, or both Draws and completed the DHQ and 4 24-HR

 
Mean dietary intakes of all individual carotenoids were higher for the DHQ than the 4 24-HR for women and men, but only significantly so for ß-cryptoxanthin intake in women (Table 3). In both women and men, none of the vitamin E estimates differed between instruments.


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TABLE 3 Geometric means and 95% CI for dietary carotenoids and tocopherols from women and men who had blood drawn at Draw 1 and completed the DHQ and 4 24-HR

 
Serum–diet correlations were moderately strong for {alpha}-carotene and ß-cryptoxanthin for both dietary instruments, particularly in men (r = 0.4–0.6, P < 0.01) (Table 4). Correlations for ß-carotene were modest in general and strongest for the 4 24-HR in men (r = 0.4, P < 0.05). Correlations for lutein + zeaxanthin were moderately strong only for the 4 24-HR in women (r = 0.5, P < 0.001) and only for the DHQ in the fully adjusted models in men (r = 0.4, P < 0.01). Serum lycopene was more highly correlated with intake in men (r = 0.4 to 0.5, P < 0.01) than in women regardless of instrument or adjustment with covariates. Correlations changed when a summary measure of total ß-carotene equivalents was related to serum {alpha}- and ß-carotene. Compared with only {alpha}-carotene from food, correlations of total ß-carotene equivalents with serum {alpha}-carotene were stronger for the 4 24-HR for women but weaker for the DHQ for men. Compared with only ß-carotene from food, correlations between total ß-carotene equivalents and serum ß-carotene were slightly stronger for both instruments in women and in men.


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TABLE 4 Spearman rank correlation coefficients between serum and dietary carotenoids and tocopherols from women and men who had blood drawn at Draw 1 and completed the DHQ and 4 24-HR

 
Serum {alpha}-tocopherol correlated poorly with dietary {alpha}-tocopherol for both instruments in women (Table 4). In men, serum {alpha}-tocopherol was moderately correlated (0.4–0.5, P < 0.01) with dietary {alpha}-tocopherol from both instruments after adjustment for covariates. In general, serum {gamma}-tocopherol did not correlate with dietary {gamma}-tocopherol. However, in women and in men, serum {alpha}-tocopherol was strongly and positively correlated (r = 0.7, P < 0.001) and serum {gamma}-tocopherol was strongly and inversely correlated (r = –0.6 to –0.7, P < 0.001) with total vitamin E from food and dietary supplements for both instruments.

With the exception of the dietary tocopherols, adjustment for energy, BMI, smoking status, serum cholesterol, and serum triacylglycerol did not appreciably change the respective values or the significance of the correlations. We also evaluated the serum–diet correlations within strata of energy intake, BMI, and smoking status. For the DHQ, serum–diet correlations for the carotenoids and tocopherols differed the most between never and former or current smokers and, in some cases, the correlations were higher in women who were former or current smokers. For the 4 24-HR, serum–diet correlations for the carotenoids tended to be lower in women who underreported energy, were overweight, or smoked. In these women, serum–diet correlations for the tocopherols tended to be higher than in women with adequate energy intake, normal weight, or who never smoked; but correlations for total vitamin E were similar between all comparison groups. The effects of energy intake, weight, or smoking on serum–diet correlations were not consistent for either instrument in men.

Most validity coefficients for carotenoids and tocopherols estimated from the DHQ, the 4 24-HR, and serum were strong (Table 5). In men, the validity coefficients for estimating carotenoids from the DHQ were all >0.5, with the exception of lycopene, and were highest for {alpha}-carotene, ß-cryptoxanthin, and lutein + zeaxanthin compared with values for the 4 24-HR or serum. Similarly, in women, most validity coefficients for carotenoids estimated from the DHQ were >0.5, although they tended to be lower than the respective values in men. To note, the validity coefficient for lutein + zeaxanthin from the DHQ was lowest in women but highest in men compared with the 4 24-HR and serum. In both women and men, validity coefficients for the individual tocopherols and total vitamin E tended to be highest for the DHQ. The ranges of most validity coefficients were wide, but most coefficients had upper limits >0.5.


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TABLE 5 Validity coefficients and their ranges for carotenoids and tocopherols estimated from the DHQ, 4 24-HR, and serum from Draw 1

 
Analyses repeated among participants who had blood drawn at time 2 or at both times showed similar results for the crude and adjusted serum–diet correlations and validity coefficients presented for the analytic sample with blood drawn at time 1.


    Discussion
 TOP
 ABSTRACT
 Introduction
 Methods
 Results
 Discussion
 LITERATURE CITED
 
In this sample of EATS participants, mean intakes of carotenoids and tocopherols from the DHQ were similar to those from the 4 24-HR collected over the course of 1 y. Mean intakes were also comparable to national data (17,18). Mean concentrations of serum carotenoids and tocopherols of EATS participants were also comparable to national data, although EATS participants had lower serum ß-cryptoxanthin and higher serum lycopene concentrations than adult participants in NHANES III (mean cryptoxanthin was 0.17 µmol/L and lycopene was 0.44 µmol/L (19). These differences may be related to the testing laboratory, different isomers measured, or to inherent differences between the NHANES and EATS participants.

The EATS data showed modest to strong serum–diet correlations for the carotenoids with provitamin A activity (i.e., {alpha}-carotene, ß-carotene, ß-cryptoxanthin), low to modest correlations for lycopene, and very low correlations for lutein. The provitamin A carotenoids and lycopene are found primarily in a few foods included on the DHQ (i.e., carrots and spinach for {alpha}- and ß-carotene, orange juice and oranges for ß-cryptoxanthin, tomatoes and tomato products for lycopene). Lutein and zeaxanthin are found primarily in green leafy vegetables but also in a variety of other foods. Intake of lutein and zeaxanthin is more difficult to quantify because of known limitations of food composition tables for these carotenoids (20). Other studies have also reported similar serum–diet correlations for lutein (2124).

The EATS data showed weak serum–diet correlations for the individual tocopherols, which may be due to measurement error from: 1) underreporting of energy and fat intake, 2) poor assessment of fats and oils added during food preparation, 3) uncertainty regarding the types of fats and oils consumed, and 4) high variability of vitamin E content in the food composition databases (19). In contrast, the EATS data showed strong positive correlations for total vitamin E from both instruments with serum {alpha}-tocopherol but strong inverse correlations with serum {gamma}-tocopherol. Like other studies (19,25), these results are consistent with the knowledge that supplemental vitamin E is in the form of {alpha}-tocopherol, which overwhelms amounts in the typical diet (400 mg vs. 8 mg/d from food on average), and that after absorption, {alpha}-tocopherol is preferentially resecreted by the liver into circulation, whereas {gamma}-tocopherol and other tocopherols are excreted.

In theory, a FFQ should better estimate typical intake, especially for foods infrequently consumed that may be rich sources of a single nutrient (e.g., yams for ß-carotene), compared with single or multiple days of dietary intake when consumption might be missed on recording days. The findings from EATS support this theory to some degree but seem to be nutrient dependent. The DHQ had higher correlations for serum {alpha}-carotene, ß-cryptoxanthin (men only), lutein (men only), and {alpha}-tocopherol, whereas the 4 24-HR had higher correlations for ß-carotene, total ß-carotene equivalents, lycopene (men only), and {gamma}-tocopherol. The magnitude of the correlations was often comparable between instruments. We observed no clear advantage of 4 24-HR, suggesting that the DHQ is adequate for large epidemiologic studies that may not be able to include multiple recalls.

For comparison, we identified 7 studies that reported serum–diet correlations for carotenoids and/or tocopherols with at least 1 FFQ and multiple recalls (2632) and another 6 studies with multiple food records (21,3337). The serum–diet correlations reported in these studies varied, and no particular dietary method produced consistently stronger correlations for individual carotenoids or tocopherols. Some research (38) suggests that weighed food records or diet diaries may produce more valid nutrient estimates than FFQ or multiple recalls. However, the serum–diet correlations for the provitamin A carotenoids and total vitamin E from the DHQ and from the 4 24-HR were as strong as those reported in studies that used multiple weighed food records or diet diaries (21,3337). In EATS and other studies, the variability in the serum–diet correlations by instrument likely depends on the population group, their particular dietary intakes, the proximity of blood collection to administration of the 4 24-HR, and other unknown factors. Combining both instruments may provide better estimates of usual intake than using either instrument alone (32,39).

Like other studies (40,41), the EATS data showed strong reproducibility for the individual carotenoids and tocopherols between the first and second draws, a time period of ~6 mo. In EATS, the serum–diet correlations were comparable whether serum data were from the first, second, or the mean of both draws. They were also comparable to those reported in the literature from 2 blood draws (21,31,33,34). These findings suggest that 1 blood collection within a year is likely to be representative of usual nutritional status for these nutrients, and that there is no advantage to collecting >1 blood sample for epidemiologic studies, especially because of cost and burden to the participants.

Adjustment for energy, smoking, body weight, and serum lipids, did not appreciably improve the crude serum–diet correlations for the DHQ or the 4 24-HR, except for {alpha}-tocopherol for both dietary instruments in men. Like EATS, some studies (21,28,33) showed no improvement with adjustment for energy, lipids, and covariates, whereas other studies (27,44) showed improvement for {alpha}-tocopherol but less change for individual carotenoids. In our study, underreporting appeared to only affect the serum–diet correlations for the 4 24-HR in women, resulting in lower correlations compared with women who reported adequate energy intake. For the DHQ in women and for both instruments in men, the serum–diet correlations were comparable between participants with adequate energy intake and those who may have underreported, suggesting that the DHQ ranks well even among underreporters.

Using Kaaks' method of triads (8), validity coefficients for the carotenoids and tocopherols for the DHQ were comparable to those observed for the 4 24-HR and the serum and were notably strong (>0.8) for {alpha}-carotene, ß-cryptoxanthin, lutein + zeaxanthin, and total vitamin E in men and for {gamma}-tocopherol and total vitamin E in women. In other studies (26,29,32,35,37), the validity coefficients were comparable in magnitude to the values observed in EATS. Ranges of validity coefficients in EATS were wide, leading us to conclude, similarly, that only strong associations between many of these dietary factors and disease will be detected (37).

EATS is one of the few studies that compares multiple methods of dietary assessment to a spectrum of serum carotenoids and tocopherols. The database (11) and methodology used (12) to estimate intake of individual carotenoids and tocopherols from the DHQ and the 4 24-HR were more comprehensive than previous studies (21,43,44). However, there are a variety of factors that impact direct correlations between intake and serum estimates, many of which cannot be addressed with current data. The most troubling limitation of all dietary assessment methods is measurement error, especially due to the underreporting of energy intake (45). We do not know to what degree such error impacts the measurement of individual carotenoids and tocopherols. In addition, correlated errors between a FFQ and reference method (multiple recalls or records) are likely to be much greater than observed (46), making the validity coefficients from the method of triads overestimated. An alternative approach using multiple latent variables, such as 2 biomarkers, may better satisfy assumptions about correlated errors in assessment methods (47), but these models need further exploration (48).

Even with these limitations, findings from EATS show that the DHQ performed generally as well as multiple recalls when compared with serum concentrations of carotenoids and tocopherols. The NCI DHQ produced reliable and valid estimates for dietary carotenoids and tocopherols and diet–serum correlations with strong validity coefficients, demonstrating its comparability to other FFQs for use in large epidemiologic studies of diet and health.


    ACKNOWLEDGMENTS
 
We appreciate the extensive database work by Thea Zimmerman and the assistance of Temitope Erinosho with making data tables.


    FOOTNOTES
 
1 This work was supported in part by an independent contract from the Division of Cancer Control and Population Sciences, National Cancer Institute, NIH (L.B.D.). None of the authors have conflicts of interest. Back

5 Abbreviations used: DHQ, diet history questionnaire; EATS, Eating at America's Table Study; FFQ, food frequency questionnaire; 4 24-HR, four 24-h dietary recalls. Back

Manuscript received 21 May 2006. Initial review completed 19 June 2006. Revision accepted 21 September 2006.


    LITERATURE CITED
 TOP
 ABSTRACT
 Introduction
 Methods
 Results
 Discussion
 LITERATURE CITED
 

1. Thompson FE, Subar AF. Dietary assessment methodology. In: Coulston AM, Rock CL, Monsen ER, editors. Nutrition in the prevention and treatment of disease. San Diego (CA): Academic Press, 2001.

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