Andrea M. Hutchins

Urinary Equol Excretion with a Soy Challenge: Influence of Habitual Diet

Abstract Equol is an isoflavonoid phytoestrogen produced from the soy isoflavone daidzein by gut microflora. Not all humans produce equol from daidzein, presumably due to differences in colonic bacterial populations among individuals. Previously, smaller studies reported that approximately 30% of participants excreted equol when consuming soy. The purpose of our study was to determine the prevalence of equol excreters in a larger sample and to examine what dietary components might influence the tendency to be an equol excreter. Thirty men and thirty women consumed a soy protein beverage containing 22 mg genistein and 8 mg daidzein for 4 days as a supplement to their habitual diets. The mean daily nutrient content of their habitual intakes was determined from 4-day food records. On Day 4, participants provided a 24-hour urine collection. Urinary isoflavonoid (genistein, daidzein, equol, and O-desmethy-langolensin) excretion was measured by gas chromatography-mass spectrometry. Twenty-one of the 60 participants (35%) excreted equol (> 2000 nmol/day) after 3 days of consuming the soy supplement. Daily equol excretion ranged from 2,134–20,301 nmol/day in the excreters and 21–233 nmol/day in the nonexcreters. There was no difference in equol excreter prevalence between men (43%) and women (27%). Daily excretion of daidzein, genistein, and O-desmethylangolensin was similar between equol excreters and nonexcreters and between men and women. Among the women, equol excreters consumed a significantly higher percentage of energy as carbohydrate and greater amounts of plant protein and dietary fiber, both as soluble and insoluble fiber compared to nonexcreters. Such differences were not observed in the men, who overall had significantly higher fiber intakes than the women. These data suggest that, among women, dietary fiber or other components of a high-fiber diet may promote the growth and/or the activity of bacterial populations responsible for equol production in the colon.

Urinary isoflavonoid phytoestrogen and lignan excretion after consumption of fermented and unfermented soy products

OBJECTIVE To compare the effects of consumption of fermented and unfermented soy products on excretion of urinary isoflavonoid phytoestrogens and lignans in healthy men. DESIGN A randomized, crossover trial consisting of two 9-day feeding periods following 5 days of baseline data collection. SUBJECTS Healthy men, aged 20 to 40 years, were recruited from the University of Minnesota Twin Cities community. Of the 22 subjects who began the study, 17 completed all feeding periods. INTERVENTIONS Fermented soy product (112 g tempeh) or unfermented soy (125 g soybean pieces) was consumed during each controlled feeding period. MAIN OUTCOME MEASURE Urine samples collected while subjects consumed their habitual diets and on the last 3 days of each feeding period were analyzed for isoflavonoid and lignan content by isotope dilution gas chromatography-mass spectrometry. STATISTICAL ANALYSIS PERFORMED Comparisons of isoflavonoid and lignan excretion were analyzed using the general linear model procedure. Orthogonal contrasts were used to determine treatment differences of interest. RESULTS Urinary excretion of isoflavonoids (equol, O-desmethylangolensin [O-DMA], daidzein, genistein) was higher and excretion of lignans (enterodiol, enterolactone) was lower when subjects consumed soy-supplemented diets than when they consumed their habitual diets (P < .05). Urinary isoflavonoid excretion and lignan excretion were similar when subjects consumed tempeh and soybean pieces diets; however, recovery of daidzein and genistein was significantly higher when subjects consumed the tempeh diet than when they consumed the soybean pieces diet (P < .002). When fed soy, 5 of 17 subjects excreted high amounts of equol. These five subjects tended to excrete less O-DMA and daidzein than the 12 subjects who excreted low amounts of equol (P < .06). CONCLUSIONS Fermentation of soy decreased the isoflavone content of the product fed but increased the urinary isoflavonoid recovery. This finding suggests that fermentation increases availability of isoflavones in soy.

Effect of Flaxseed Consumption on Urinary Estrogen Metabolites in Postmenopausal Women

Flaxseed, the richest known source of plant lignans, has been shown to have chemoprotective effects in animal and cell studies. Some of its effects may be mediated through its influence on endogenous hormone production and metabolism. Two competing pathways in estrogen metabolism involve production of the 2-hydroxylated and 16 alpha-hydroxylated metabolites. Because of the proposed differences in biological activities of these metabolites, the balance of the two pathways has been used as a biomarker for breast cancer risk. We examined the effects of flaxseed consumption on urinary estrogen metabolite excretion in postmenopausal women. Twenty-eight postmenopausal women were studied for three seven-week feeding periods in a randomized crossover design. During the feeding periods, subjects consumed their usual diets plus ground flaxseed (0, 5, or 10 g/day). Urinary excretion of the estrogen metabolites 2-hydroxyestrogen (2-OHEstrogen) and 16 alpha-hydroxyestrone (16 alpha-OHE1) as well as their ratio, 2/16 alpha-OHE1, was measured by enzyme immunoassay. Flaxseed supplementation significantly increased urinary 2-OHEstrogen excretion (p < 0.0005) and the urinary 2/16 alpha-OHE1 ratio (p < 0.05) in a linear, dose-response fashion. There were no significant differences in urinary 16 alpha-OHE1 excretion. These results suggest that flaxseed may have chemoprotective effects in postmenopausal women.

Vegetables, fruits, and legumes: Effect on urinary isoflavonoid phytoestrogen and lignan excretion

OBJECTIVE To compare the effect of vegetable, fruit, and legume consumption on urinary isoflavonoid phytoestrogen and lignan excretion. DESIGN After 4 days of data collection, during which subjects consumed their habitual diets, subjects were randomly placed on four 9-day controlled experimental diets with each subject receiving each diet in a random order. SUBJECTS Seven men and three women, aged 20 to 35 years, were recruited from the University of Minnesota Twin Cities community. INTERVENTIONS All subjects consumed four experimental diets in an assigned random order: a controlled basal diet, a legume/allium diet (containing garbanzo beans, garlic, and onions), and diets low or high in vegetables and fruits (containing apples, pears, potatoes, and carrots). MAIN OUTCOME MEASURES Urine samples that were collected while subjects consumed their habitual diets and during the last 3 days of each feeding period were analyzed for isoflavonoid and lignan content using isotope dilution gas chromatography-mass spectrometry. STATISTICAL ANALYSIS PERFORMED The effect of vegetable and fruit intake on urinary isoflavonoid and lignan excretion was analyzed using the general linear model procedure. Post hoc comparisons were made using Duncans multiple range test. RESULTS Subjects excreted more of the lignan enterodiol on the high vegetable/fruit diet compared with the basal and legume/allium diets (P = .03); more of the isoflavonoids O-desmethylangolensin (O-DMA), genistein, and sum of isoflavonoids on the legume/allium diet compared with the other controlled diets (P < .05); and more of the isoflavan equol on the basal and legume/allium diets compared with the high vegetable/fruit diet (P < .01). Subjects who excreted higher levels of equol on the basal and legume/allium diets also consumed more of the milk-based pudding provided as part of the controlled diets. CONCLUSIONS Urinary lignan and isoflavonoid excretion changed in response to alterations in vegetable, fruit, and legume intake under controlled dietary conditions.

Flaxseed Consumption Influences Endogenous Hormone Concentrations in Postmenopausal Women

Lignans, similar in structure to endogenous sex steroid hormones, may act in vivo to alter hormone metabolism and subsequent cancer risk. The objective of this study was to examine effects of dietary intake of a lignan-rich plant food (flaxseed) on serum concentrations of endogenous hormones and binding proteins (estrone, estrone sulfate, 17β-estradiol, sex hormone-binding globulin, progesterone, prolactin, dehydroepiandrosterone sulfate, dehydroepiandrosterone, androstenedione, testosterone, and free testosterone) in postmenopausal women. This randomized, crossover trial consisted of three seven-week feeding periods, during which 28 postmenopausal women, aged 52-82 yr, consumed their habitual diets plus 0, 5, or 10 g of ground flaxseed. Serum samples collected during the last week of each feeding period were analyzed for serum hormones using standard diagnostic kits. The flaxseed diets significantly reduced serum concentrations of 17β-estradiol by 3.26 pg/ml (12.06 pmol/l) and estrone sulfate by 0.09 ng/ml (0.42 nmol/l) and increased prolactin by 1.92 μg/l (0.05 IU/ml). Serum concentrations of androstenedione, estrone, sex hormone-binding globulin, progesterone, testosterone, free testosterone, dehydroepiandrosterone, and dehydroepiandrosterone sulfate were not altered with flaxseed feeding. In this group of postmenopausal women, consuming flaxseed in addition to their habitual diets influenced their endogenous hormone metabolism by decreasing serum 17β-estradiol and estrone sulfate and increasing serum prolactin concentrations.

Influence of soybean processing, habitual diet, and soy dose on urinary isoflavonoid excretion

In an attempt to explain the wide individual variation seen in urinary isoflavonoid phytoestrogen excretion, we conducted a series of 3 human feeding studies: a large cross-sectional study of equol production in humans with a soy challenge, a comparison of phytoestrogen metabolism when subjects consumed fermented and unfermented soy products, and a dose-response study of urinary isoflavonoid excretion at the low end of soy consumption. All studies were conducted in young, healthy humans. Urinary isoflavonoids were measured by isotope-dilution gas chromatography-mass spectrometry. Similar to results from other studies, 35% of screened subjects (30 men and 30 women) excreted equol (>2000 nmol/d). In women, equol excretion was associated with higher intake of dietary fiber and carbohydrate. Fermentation of soy decreased the isoflavone content of the product fed but increased the urinary isoflavonoid recovery, suggesting that fermentation increases availability of isoflavones in soy. When soy-protein powder was fed at 0, 5, 10, and 20 g/d (0-36 mg isoflavones), there was a linear dose response of urinary isoflavonoid excretion to soy consumption that did not differ between subjects with high and low equol excretion. These results suggest that equol excretion may be related to the fermentable carbohydrate content of the diet; additional study is needed. Processing of soy affects isoflavone metabolism and must be considered in recommending exposure to isoflavones from soyfoods. Although optimal isoflavone exposure for disease protection has not been determined, urinary isoflavonoid excretion appears linear at low-to-moderate soy consumption.

Types of Dietary Fat and Soy Minimally Affect Hormones and Biomarkers Associated With Breast Cancer Risk in Premenopausal Women

Fourteen premenopausal women participated in a randomized, crossover controlled feeding study of three diets, each two menstrual cycles long. We compared a high saturated fat Western diet (control diet) with two other diets: the control diet plus soy protein (soy diet) and the control diet with polyunsaturated fat (PUFA diet) replacing most of the saturated fat. We measured reproductive and serum hormones, urinary estrogen metabolites and isoflavonoids, and menstrual cycle length. In the follicular phase, prolactin concentrations significantly decreased by 3.6 μg/dl (P = 0.047), follicle-stimulating hormone concentrations slightly increased by 0.1 IU/l (P = 0.076), and cortisol concentrations slightly decreased by 81.8 nmol/l (P = 0.088) with the PUFA diet vs. the control diet. The soy diet slightly increased menstrual cycle length by 1.8 ± 0.7 days (P = 0.088) and significantly increased (P < 0.0001) urinary isoflavonoid excretion. These well-controlled diets did not affect serum estrogens or urinary estrogen metabolites, suggesting that type of fat or consumption of soy with a high saturated fat diet may not alter breast cancer risk by these mechanisms.

Effect of Flaxseed and Wheat Bran on Serum Hormones and Lignan Excretion in Premenopausal Women

Objective: To examine the effects of flaxseed consumption (a lignan-rich plant food) alone and in combination with wheat bran on serum hormones and urinary lignan excretion in premenopausal women. Methods: Sixteen subjects were studied for four feeding treatments lasting two menstrual cycles each in a randomized, crossover design. During each treatment, subjects consumed their habitual diets supplemented with baked goods containing no flaxseed or wheat bran, 10 g of flaxseed, 28 g of wheat bran, or 10 g of flaxseed plus 28 g of wheat bran/day. Serum samples collected during the mid-luteal phase of the second menstrual cycle of each diet treatment were analyzed for serum hormones and sex hormone binding globulin. Urine samples collected during the same time period were analyzed for urinary lignan excretion. Results: There were no changes in serum hormone concentrations or sex hormone binding globulin on any of the treatments. Urinary lignan excretion significantly increased on diet treatments that included flaxseed, but inclusion of wheat bran did not significantly alter lignan excretion. Conclusions: Urinary lignan excretion increased with flaxseed consumption, but serum hormones did not change in this group of premenopausal women. Consumption of wheat bran with flaxseed did not alter urinary lignan excretion associated with flaxseed consumption.

Dietary Estrogens Influence Urinary Lignan Excretion in a Dose-Dependent Manner in Post-Menopausal Women

Abstract Dietary estrogens, such as lignans, are similar in structure to endogenous sex steroid hormones and may act in vivo to alter hormone metabolism and subsequent cancer risk. The objective of this study was to examine the effect of dietary intake of a lignan-rich plant food (flaxseed) on urinary lignan excretion in post-menopausal women. This randomized, crossover trial consisted of three 7-week feeding periods during which 31 healthy post-menopausal women, aged 52-82, recruited from the Monastery of the Sisters of the Order of St. Benedict in St. Joseph, MN consumed their habitual diets plus 0, 5 or 10 grams of ground flaxseed. Urine samples collected for 2 consecutive days during the last week of each feeding period were analyzed for lignan content (enterodiol, enterolactone, matairesinol) by isotope dilution gas chromatography/mass spectrometry. Compared to the 0g flaxseed diet, consuming 5 or 10g of flaxseed increased excretion of enterodiol by 3.73 (2.71-5.13) pmol/mg creatinine (95% confidence interval [CI]), p=0.0001 and 8.71 (6.32-11.99) pmol/mg creatinine (95% CI), p=0.0001, respectively, and increased excretion of enterolactone by 7.15 (5.48-9.12) pmol/mg creatinine (95% CI), p=0.0001 and 16.18 (12.41 -21.09) pmol/mg creatinine (95% CI), p=0.0001, respectively. Excretion of matairesinol was not significantly altered with flaxseed feeding. Consuming flax, a significant source of dietary estrogens, in addition to their habitual diets increased excretion of enterodiol and enterolactone, but not matairesinoi, in a dose-dependent manner in this group of post-menopausal women. Urinary excretion of lignan metabolites is a dose-dependent biomarker of flaxseed intake within the context of a semi-controlled diet.