Women's Healthy Lifestyle Project: A randomized clinical trial: results at 54 months.

BACKGROUND: The Women's Healthy Lifestyle Project Clinical Trial tested the hypothesis that reducing saturated fat and cholesterol consumption and preventing weight gain by decreased caloric and fat intake and increased physical activity would prevent the rise in LDL cholesterol and weight gain in women during perimenopause to postmenopause. METHODS AND RESULTS: There were 275 premenopausal women randomized into the assessment only group and 260 women into the intervention group. The mean age of participants at baseline was 47 years, and 92% of the women were white. The mean LDL cholesterol was 115 mg/dL at baseline, and mean body mass index was 25 kg/m(2). The follow-up through 54 months was excellent. By 54 months, 35% of the women had become postmenopausal. At the 54-month examination, there was a 3.5-mg/dL increase in LDL cholesterol in the intervention group and an 8.9-mg/dL increase in the assessment-only group (P:=0.009). Weight decreased 0.2 lb in the intervention and increased 5.2 lb in the assessment-only group (P:=0.000). Triglycerides and glucose also increased significantly more in the assessment-only group than in the intervention group. Waist circumference decreased 2.9 cm in the intervention compared with 0.5 cm in the assessment-only group (P:=0.000). CONCLUSIONS: The trial was successful in reducing the rise in LDL cholesterol during perimenopause to postmenopause but could not completely eliminate the rise in LDL cholesterol. The trial was also successful in preventing the increase in weight from premenopause to perimenopause to postmenopause. The difference in LDL cholesterol between the assessment and intervention groups was most pronounced among postmenopausal women and occurred among hormone users and nonusers.

Influence of the perimenopause on cardiovascular risk factors and symptoms of middle-aged healthy women.

OBJECTIVE: To determine the changes in cardiovascular risk factors and psychological and physical symptoms that occur during the perimenopause. DESIGN: Cohort study of 541 healthy middle-aged premenopausal women followed up through the menopause. SETTING: General community. PARTICIPANTS: After a baseline evaluation taken at study entry, 152 women ceased menstruating for 3 months (not due to surgery) and were not using hormone replacement therapy, and were reevaluated in a similar protocol (perimenopausal examination); 105 of the 152 were evaluated a third time when they had ceased menstruating for 12 months and were not using hormone replacement therapy (postmenopausal examination). One hundred nine premenopausal women who were repeatedly tested constituted a comparison group. MAIN OUTCOME MEASURES: Levels of lipids and lipoproteins, triglycerides, fasting glucose and insulin, blood pressure, weight, height, and standardized measures of psychological symptoms. RESULTS: Women who became perimenopausal showed increased levels of cardiovascular risk factors, which were similar in magnitude to those experienced by the comparison group of premenopausal women. Perimenopausal women reported a greater number of symptoms, especially hot flashes, cold sweats, joint pain, aches in the skull and/or neck, and being forgetful; reports of hot flashes at the perimenopausal examination were associated with low concentrations of serum estrogens. Menopausal status was not associated with depressive symptoms. Perimenopausal women who became postmenopausal showed a decline in the level of high-density lipoprotein-2-cholesterol (means, 0.53 to 0.43 mmol/L [20.6 to 16.7 mg/dL]) and a gradual increase in the level of low-density lipoprotein cholesterol (means, 3.14 to 3.33 mmol/L [121.3 to 128.8 mg/dL]), whereas symptom reporting declined. CONCLUSIONS: During mid-life, women experience adverse changes in cardiovascular risk factors and a temporary increase in total number of reported symptoms, with no change in depression. Preventive efforts to reduce the menopause-induced increase in cardiovascular risk factors should begin early in the menopausal transition.

Weight gain at the time of menopause.

We studied prospectively the weight change and the effect of weight change on changes in coronary heart disease risk factors in a population-based sample of 485 middle-aged women. All women were studied first in 1983 to 1984, when they were premenopausal and aged 42 to 50 years, and then restudied in 1987. Women gained an average of 2.25 +/- 4.19 kg during this 3-year period; 20% of women gained 4.5 kg or more, and only 3% lost 4.5 kg or more. There were no significant differences in weight gain of women who remained premenopausal and those who had a natural menopause (+2.07 kg vs +1.35 kg). Weight gain was significantly associated with increases in blood pressure and levels of total cholesterol, low-density lipoprotein cholesterol, triglycerides, and fasting insulin. Weight gain is thus a common occurrence for women at the time of menopause and is related to the changes in coronary heart disease risk factors observed during this period. Efforts to lose weight or to prevent weight gain may help to mitigate the worsening in coronary heart disease risk factors in middle-aged women.

Body composition and bone mineral density in premenopausal and early perimenopausal women.

Body composition appears to be an important determinant of bone mineral density (BMD). BMD at the femoral neck, lumbar spine, and whole-body and the whole-body soft-tissue composition were measured cross-sectionally in 334 healthy premenopausal and early perimenopausal women, aged 44-50 years, using a Hologic QDR densitometer. Correlations between lean mass and BMD at the hip, spine, and whole-body were greater (r = 0.40, r = 0.44, and r = 0.45, respectively, p < 0.0001) than those for fat mass (r = 0.19, r = 0.16, and r = 0.16, respectively, p < 0.01). There was a significant linear trend in femoral BMD from the lowest to highest category of lean mass (0.75 +/- 0.10 g/cm2, 0.80 +/- 0.10 g/cm2, and 0.86 +/- 0.09 g/cm2, p < 0.0001). Similar trends were demonstrated for spinal and whole-body density. For categories of fat mass, there was a significant linear trend at the hip (0.78 +/- 0.10 g/cm2, 0.79 +/- 0.10 g/cm2, and 0.83 +/- 0.10 g/cm2, p = 0.0106), but not at the spine or whole body. There was a 5.00% (3.62, 6.38; 95% confidence limits) difference in hip BMD per unit (standard deviation) of lean mass, while only a 0.73% (-0.66, 2.11) difference in hiP BMD per unit (SD) of fat mass. Differences in BMD were examined by categories of lean and fat mass (low, medium, high) for a total of nine possible combinations of lean and fat measures. BMD at the hip, spine, and whole-body were significantly higher in those with high lean mass than in those with low lean mass, irrespective of fat mass. Women with high lean/low fat had similar hip, spinal, and whole-body BMD as those with high lean/high fat, despite their significantly lower body weight (62.5 +/- 3.3 kg vs 85.7 +/- 5.4 kg, respectively, p < 0.0001). In premenopausal and early perimenopausal women, body weight alone may not be associated with increased bone mass unless a significant proportion of that weight is comprised of lean mass. The stronger association between lean mass and BMD than that for fat mass may be attributed to differences in determinants of lean mass, such as exercise, lifestyle factors, estrogen levels, or a combination of these factors.

Determinants of bone mineral density in older men.

Osteoporosis is a significant health problem and contributor to disability and premature mortality among older men. Incidence rates for hip fracture have stabilized in women, but continue to increase in men. A major risk factor for hip fracture is bone mineral density level. The determinants of bone mineral density (BMD) are well defined in women, but not in men. The primary goal of the current research was to increase our understanding of the determinants of BMD of the proximal femur in a large community-based sample of older male volunteers. Eligibility requirements included age of 50 years or older, ambulatory, and not having undergone a bilateral hip replacement. Five hundred twenty-three men, mean mean age 66.6 years, met all eligibility requirements and participated in the Study of Osteoporotic Risk in Men or STORM. Information on demographics, medical history, anthropometry, leisure-time and occupational physical activity, muscular strength, cigarette smoking, alcohol consumption, dietary calcium intake, and medication use (thiazide diuretics and glucocorticoids) were obtained by questionnaire, interview, and examination, BMD of the proximal femur (femoral neck, greater trochanter, and Ward's triangle) was measured by dual-energy X-ray absorptiometry using the Hologic QDR-1000 and QDR-2000. The cross-sectional determinants of BMD included age, blond hair color, current body weight, thiazide diuretic use, historical physical activity, and quadriceps strength. Several variables commonly thought to be determinants of BMD were not related to BMD in this population of older men, including current cigarette smoking, alcohol consumption, current leisure-time physical activity, dietary calcium intake, vitamin D use, and caffeine intake. However, failure to find associations among BMD and some of the potential determinants may be due to lack of statistical power. Identification of the determinants of BMD could lead to the development of interventions aimed at maximizing BMD in men and could potentially decrease the risk of hip fractures.

Lipoprotein(a) and peripheral atherosclerosis in older adults.

As part of an ancillary study to the Systolic Hypertension in the Elderly Program, carotid and lower extremity arterial disease (LEAD) were evaluated in 369 subjects, 186 with a systolic blood pressure (SBP) > or = 160 mmHg, and 183 with SBP < 160 mmHg. Both groups had a diastolic blood pressure (DBP) < 90 mmHg. Internal carotid stenosis was identified by Doppler and LEAD was assessed using the ankle to arm systolic blood pressure ratio, commonly called the ankle/arm index (AAI). Lp(a) values were obtained from frozen sera and values > or = 20 mg/dl were considered elevated. Rates of carotid stenosis were 24% among those with an Lp(a) > or = 20 mg/dl and 14% among those with an Lp(a) level < 20 mg/dl (P = 0.020). The relationship between Lp(a) and LEAD was even stronger. Those with an Lp(a) > or = 20 mg/dl had a 36% prevalence of a low AAI vs 14% among those with a Lp(a) level < 20 mg/dl (P < 0.001). Lp(a) values were also associated with the severity of LEAD. Controlling for other risk factors did not reduce the association between either LEAD or carotid stenosis and an Lp(a) > or = 20 mg/dl. Thus, Lp(a) appears to be independently associated with peripheral atherosclerosis in older adults, both men and women. The relationship is particularly strong for atherosclerotic disease of the lower extremities.

Psychological, biological and health behavior predictors of blood pressure changes in middle-aged women.

A long-standing hypothesis is that feelings of anger and anxiety increase the risk for essential hypertension. Most studies examining this hypothesis have been cross-sectional in design or undertaken with men only. We tested this hypothesis along with determination of the other behavioral and biological predictors of increases in systolic (SBP) and diastolic (DBP) blood pressure from baseline to a follow-up examination 3 years later in a prospective study of 468 middle-aged women whose blood pressure at the baseline examination was less than 140/90 mmHg. Analyses showed that increases in the Spielberger Trait Anger Scale between the baseline and 3-year follow-up examination, as well as Framingham Tension scores (a measure of anxiety) at baseline, independently predicted an increase in SBP (P less than 0.01). Other factors that independently predicted an increase in SBP were baseline fasting insulin, parental history of hypertension and increases in body mass index and in alcohol intake across the 3 years of follow-up. Increases in the Spielberger Trait Anger Scores independently predicted increases in DBP (P less than 0.02), as did black race, increases in body mass index and hematocrit and decreases in potassium intake. Although menopausal status and hormone replacement therapy were unrelated to changes in blood pressure, postmenopausal women on hormone replacement therapy did show significant increases in DBP in the univariate analysis. Anxiety at baseline, along with parental history of hypertension, baseline fasting insulin and baseline body mass index, predicted a later onset of hypertension, i.e. on pharmacologic treatment for hypertension, in the univariate analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Genetic determination of coagulation factor VIIc levels among healthy middle-aged women.

A recent study (1) reported variation among men in clotting factor VIIc levels is associated with a genetic polymorphism detected by the restriction enzyme Msp I. The present study determined the Msp I genotype (Arg353, Gln353 alleles) for 189 women (mean age 53) who were subjects in the Healthy Women Study, a population study of CHD risk factor change at menopause. Women with the Arg/Arg genotype (n = 147) had an 16% higher (geometric) mean FVIIc level than those with the Arg/Gln (n = 41) genotype (1.21 vs 1.04 U/ml, p < 0.01), while the one subject with the Gln/Gln genotype had an FVIIc level of 1.00 U/ml. These results are consistent with those previously found in healthy men (1). In addition, women carrying the Gln allele did not exhibit the elevation in FVIIc with menopause and use of hormone therapy found among those with the Arg allele, suggesting that genotype may modify the observed rise in factor VIIc at menopause. Possibly because of the small sample size this interaction did not reach conventional levels of statistical significance. Results of multiple linear regression analyses controlling for age, hormone use, obesity, (ln) triglyceride levels, and family history of CHD found FVIIc levels to be significantly (p < 0.001) related to genotype. Thus, genotype appears to be a major determinant of FVIIc levels among women.

Biobehavioral aspects of menopause: lessons from the Healthy Women Study.

The Healthy Women Study is an ongoing natural history study of the menopause in a sample of relatively healthy women. We report data from this paper on the behavioral and biological changes that occur during the transition from pre- to postmenopausal status and argue for the importance of behavioral change interventions to prevent or attenuate some of the adverse changes due to the menopause.

Epidemiologic studies of menopause: changes in risk factors and disease.

There have been important studies of changes in risk factors and psychosocial variables during peri- and postmenopause. Most of the studies have been done in whites. Studies have clearly documented changes in behavior and biological variables related to menopause. The most critical questions bear on the interrelationships between sex steroid hormone levels, life-styles, including diet, exercise, alcohol consumption, obesity, and changes in key risk factors that are associated with the major causes of morbidity and mortality among postmenopausal women. The best study designs should be longitudinal and include frequent, accurate, and reproducible measurements of biological and psychosocial variables. Importantly, studies should be done in heterogeneous populations. The most critical variables may be measures of the degree of obesity and fatness, diet, and exercise and their relationship to hormonal changes occurring during the peri- and postmenopausal period.

Hemostatic factors according to menopausal status and use of hormone replacement therapy.

The rise in cardiovascular disease (CVD) risk after menopause may be reduced by hormone replacement therapy (HRT) although the mechanism is unclear. Because little is known about the potential role of hemostatic factors, fibrinogen level and other coagulation parameters were measured in a study on the change in CVD risk factors through the climacteric (the Healthy Women Study). Of 239 subjects measured to date, 32 taking aspirin or other medications thought to alter coagulation were excluded from analyses. Results (adjusted for age and obesity) showed that women taking HRT had lower plasma concentrations of fibrinogen and higher levels of plasminogen and factor VIIc than did postmenopausal subjects not taking HRT. Pre- as compared with postmenopausal women had lower plasma levels of fibrinogen, factor VIIc, and antithrombin III. Adjusting for cigarette smoking did not change the findings. Thus, among women aged 49 to 55, selected hemostatic measures varied (within normal ranges) by menopausal status and were altered by HRT. These findings generally support a hypothesis of hemostatic change contributing to the increase of CVD after menopause. The fact that subjects taking HRT showed no increase in fibrinogen relative to premenopausal women is consistent with an observed decreased risk of CVD among women taking HRT, while the implication of an elevation in factor VIIc among these women is uncertain.

Prevalence of thyroid antibodies among healthy middle-aged women. Findings from the thyroid study in healthy women.

Autoimmune thyroiditis is the most common cause of subclinical hypothyroidism in North America, is more common in women than men, and is a risk factor for the development of coronary heart disease (CHD). We measured thyroid-stimulating hormone (TSH) and two thyroid antibodies, thyroid peroxidase and thyroglobulin, in stored sera of the participants (aged 44 to 54 years) of the Healthy Women Study. We selected 254 samples from the premenopausal baseline examination in 1983 to 1985 and from a follow-up examination that occurred an average of 5.7 years later (range, 3 to 7.7 years). At follow-up, 95 women remained premenopausal, 98 had ceased menstruating for at least 12 months, and 61 were taking postmenopausal hormone therapy. Overall, the prevalence of the thyroid antibodies in this healthy population was high at both time points (21 to 26%). Women with antibodies had higher TSH concentrations than did those with no antibodies (2.68 +/- 1.3 versus 1.51 +/- .73 mU/L, P < 0.001); this relationship was statistically significant even after excluding those with subclinical hypothyroidism (TSH > 6.0 mU/L). TSH and antibody levels did not differ by menopausal status or hormone therapy use at follow-up. Given the high prevalence of thyroid antibodies among healthy middle-aged women, long-term follow-up is warranted to ascertain whether the presence of antibodies is associated with subsequent excess risk of disease, in particular, CHD.

Current estrogen-progestin and estrogen replacement therapy in elderly women: association with carotid atherosclerosis. CHS Collaborative Research Group. Cardiovascular Health Study.

The cardioprotective effects of combined estrogen/progestin replacement therapy have been questioned. Therefore, we have compared carotid arterial wall thickening and the prevalence of carotid stenosis in elderly women (> or = 65 years old) currently using replacement estrogen/progestins (E + P) with arterial pathology and its prevalence in women using unopposed estrogens (E). This cross-sectional study used baseline data from all 2962 women participating in the Cardiovascular Health Study, a population-based study of coronary heart disease and stroke in elderly adults. Users of hormone replacement therapy (HRT) were categorized as never (n = 1726), past (n = 787), current E (n = 280), or current E + P (n = 73). Maximal intimal-medial thicknesses of the internal and common carotid arteries and stenosis of the internal carotid arteries were measured by ultrasonography. Current E + P users resembled current E users in most respects, although some lifestyle factors were more favorable among E + P users. Current E + P use and current E use (as compared with no use) were associated with smaller internal carotid wall thicknesses (-0.22 mm; P = 0.003; and -0.09 mm; P = 0.05, respectively) and smaller common carotid wall thicknesses (-0.05 mm; P = 0.03; and -0.02 mm; P = 0.1, respectively) and lower odds ratios (OR) for carotid stenosis (> or = 1% vs. 0%); OR = 0.61; 95% confidence interval [CI]: 0.36 to 1.01; and OR = 0.91, 95% CI: 0.67 to 1.24, respectively), after adjustment for current lifestyle and risk factors. When both groups of current HRT users were compared, there were no significant differences in carotid wall thicknesses or prevalence of carotid stenosis. For this sample of elderly women, both current E + P therapy and current E therapy were associated with decreased measures of carotid atherosclerosis. These measures did not differ significantly between the two groups of HRT users.

Coffee consumption: relationship to blood lipids in middle-aged women.

BACKGROUND: There is controversy about the relationship between coffee intake and blood lipids in women and many studies fail to control for potential confounding factors. METHODS: In 1983 and 1984, 541 randomly selected premenopausal women were recruited from a list of licenced drivers, aged 42-50, within selected areas in Allegheny County, Pennsylvania. They were invited to participate in a 5-year study of biological and behavioural changes associated with menopause. At baseline and the first follow-up visit, lipoprotein, lipid and apoprotein levels, physical activity, dietary intake and coffee consumption were measured. Pearson correlations between coffee consumption and the various lipid values were calculated. Multivariate analyses, which controlled for the potential confounding effects of body mass index, alcohol intake, percent of calories from carbohydrate and fat, number of cigarettes smoked per day, physical activity level, and age in months, were also employed to investigate the relationships. RESULTS: Mean coffee consumption for the group was 3.35 cups of coffee per day at baseline and 3.02 cups per day at follow-up. Results of analyses of correlation between coffee consumption and blood lipids as well as multiple regressions to control for menopausal status at follow-up were non-significant (P > 0.05) for all blood lipid values with the exception of triglycerides, which were inversely related to coffee consumption at follow-up. CONCLUSIONS: These results do not support a relationship between coffee consumption and lipoprotein, lipid or apoprotein levels with the exception of an inverse relationship between coffee consumption and triglyceride level at follow-up.

Estrogen replacement therapy and MRI-demonstrated cerebral infarcts, white matter changes, and brain atrophy in older women: the Cardiovascular Health Study.

OBJECTIVE: We studied the relationship between the use of estrogen replacement therapy (ERT) and cerebral magnetic resonance imaging (MRI) abnormalities among older women. DESIGN: A population-based prospective study (Cardiovascular Health Study). SETTING: Four regions in the United States. PARTICIPANTS: A total of 2133 (62.9% of the eligible) women aged 65 to 95 years (mean age 74.8), on whom MRI was performed in 1992-1994. MEASUREMENTS: Presence of global brain atrophy, white matter changes, small infarct-like lesion (ILL) (<3 mm), MRI infarcts (> or =3 mm, mostly small and asymptomatic), and cognitive function as measured by Mini-Mental State Exam (MMSE), and by ERT use (current/past/never), adjusted for a number of socioeconomic, lifestyle, and reproductive covariates. RESULTS: Current use of ERT was reported by 15% and past use by another 23% of participants; 35% of all women had MRI infarcts. The prevalence of MRI infarcts did not differ in current or past users from those who had never used ERT (nonusers). Bifrontal distance, the largest distance between frontal horns, and the size of ventricles were larger among current ERT users compared to past users or nonusers (P (trend) = .01), adjusted for all other covariates, but no dose-response relationship to current or past ERT use was found. Duration of estrogen use was not associated with any atrophy measure. Cortical atrophy measure, sulcal widening, or white matter disease did not differ significantly by ERT use or duration of use. Central measures of atrophy, bifrontal distance, and ventricular size were significantly associated with cognition as measured by MMSE. CONCLUSIONS: Current ERT users had much more clinically significant central atrophy than nonusers, but the implications remained unclear.

Estrogens and women's health: interrelation of coronary heart disease, breast cancer and osteoporosis.

The determinants of blood levels of estrogen, estrogen metabolites, and relation to receptors and post-transitional effects are the likely primary cause of breast cancer. Very high risk women for breast cancer can now be identified by measuring bone mineral density and hormone levels. These high risk women have rates of breast cancer similar to risk of myocardial infarction. They are candidates for SERM therapies to reduce risk of breast cancer. The completion of the Women's Health Initiative and other such trials will likely provide a definite association of risk and benefit of both estrogen alone and estrogen-progesterone therapy, coronary heart disease, osteoporotic fracture, and breast cancer. The potential intervention of hormone replacement therapy, obesity, or weight gain and increased atherogenic lipoproteinemia may be of concern and confound the results of clinical trials. Estrogens, clearly, are important in the risk of bone loss and osteoporotic fracture. Obesity is the primary determinant of postmenopausal estrogen levels and reduced risk of fracture. Weight reduction may increase rates of bone loss and fracture. Clinical trials that evaluate weight loss should monitor effects on bone. The beneficial addition of increased physical activity, higher dose of calcium or vitamin D, or use of bone reabsorption drugs in coordination with weight loss should be evaluated. Any therapy that raises blood estrogen or metabolite activity and decreases bone loss may increase risk of breast cancer. Future clinical trials must evaluate multiple endpoints such as CHD, osteoporosis, and breast cancer within the study. The use of surrogate markers such as bone mineral density, coronary calcium, carotid intimal medial thickness and plaque, endothelial function, breast density, hormone levels and metabolites could enhance the evaluation of risk factors, genetic-environmental intervention, and new therapies.

Are job characteristics related to fibrinogen levels in middle-aged women?

This study examined whether employment status or job characteristics thought to be stressful were related to fibrinogen level in a sample of 161 healthy middle-aged women. Employed women had higher fibrinogen levels than did nonemployed women. Moreover, among employed women, those who perceived high levels of job stress or low support from their bosses had elevated fibrinogen, independent of menopausal status. Perception of low support from one's boss was related to higher fibrinogen levels only among premenopausal women or postmenopausal women who were not using hormone replacement therapy. These results are consistent with the notion that stress associated with some job characteristics influences levels of fibrinogen in women.

Diet, alcohol, and physical activity as a function of smoking status in middle-aged women.

Diet, alcohol intake, and leisure-time physical activity were compared cross-sectionally in middle-aged female smokers, ex-smokers, and never smokers and were reassessed approximately 3 years later. At initial contact, there were no group differences in total caloric intake and very few differences in nutrient intake, but alcohol intake of current smokers and ex-smokers was at least 50% greater than that of never smokers, and current smokers reported less physical activity than did ex-smokers and never smokers. Prospective findings were similar: Compared with continuing smokers, ex-smokers did not change their dietary and alcohol intake but significantly increased physical activity. Thus, some adverse behaviors associated with smoking (e.g., alcohol intake) may be due to self-selection to history of smoking, whereas other behaviors (e.g., reduced physical activity) may be more directly related to smoking itself.

Influences of natural menopause on psychological characteristics and symptoms of middle-aged healthy women.

We investigated the psychological and symptom consequences of the natural menopause in a longitudinal study of 541 initially premenopausal healthy women. All women were given an extensive evaluation at baseline. After 3 years of follow-up, 69 women ceased cycling for 12 months; another 32 women had ceased cycling and had taken hormone replacement therapy for a total of 12 months. These women were reevaluated in a clinic examination identical with the baseline examination, as were 101 age-matched premenopausal control women. Comparison among groups at the baseline and follow-up examination showed that natural menopause led to few changes in psychological characteristics, with only a decline in introspectiveness and an increase in reports of hot flashes being apparent. We conclude that natural menopause did not have negative mental health consequences for the majority of middle-aged healthy women.

Urinary markers of estrogen metabolism 2- and 16 alpha-hydroxylation in premenopausal women.

There is considerable scientific interest in whether measurement of the major estrogen metabolites 2- and 16 alpha-hydroxyestrone will shed light on the role of estrogen in the risk of breast cancer. These have been difficult to measure in large numbers because of the need for radiolabeled tracers, but a new assay is able to utilize spot urine samples. The main objective of this study was to assess the reliability of a newly developed enzyme immunoassay (EIA) for the measurement of 2- and 16 alpha-hydroxyestrone in urine samples collected from a large group of healthy premenopausal women enrolled in a clinical trial A secondary objective was to assess the impact of several factors such as body weight on the urinary estrogen metabolite ratios. The study cohort included 174 women aged 44-50, who were enrolled in the Cardiovascular Risk Factors and Menopause Trial, also referred to as the Women's Healthy Lifestyle Project (WHLP), an ongoing 5-year clinical trial of 535 premenopausal women randomized either to an intensive dietary life-style intervention group or to an assessment-only control group. Measurements of 2- and 16 alpha-hydroxyestrone showed a high intraclass correlation for blind duplicate urine samples (R = 0.94 and R = 0.80), cross-sectionally and over time (R = 0.79 and R = 0.62), in this population of healthy premenopausal women. The intervention diet (of 25% of total calories from fat) did not appear to influence the estrogen metabolite ratio. This new estrogen metabolite EIA demonstrates good reliability and thus may be appropriate for use in large epidemiologic studies of estrogen-related diseases. There was no relation between dietary fat reduction, weight loss, and increased exercise and change in the ratio among premenopausal women in this study.