Polymorphisms in the estrogen receptor alpha gene (ESR1), daily cycling estrogen and mammographic density phenotypes.

BACKGROUND: Single nucleotide polymorphisms (SNPs) involved in the estrogen pathway and SNPs in the estrogen receptor alpha gene (ESR1 6q25) have been linked to breast cancer development, and mammographic density is an established breast cancer risk factor. Whether there is an association between daily estradiol levels, SNPs in ESR1 and premenopausal mammographic density phenotypes is unknown. METHODS: We assessed estradiol in daily saliva samples throughout an entire menstrual cycle in 202 healthy premenopausal women in the Norwegian Energy Balance and Breast Cancer Aspects I study. DNA was genotyped using the Illumina Golden Gate platform. Mammograms were taken between days 7 and 12 of the menstrual cycle, and digitized mammographic density was assessed using a computer-assisted method (Madena). Multivariable regression models were used to study the association between SNPs in ESR1, premenopausal mammographic density phenotypes and daily cycling estradiol. RESULTS: We observed inverse linear associations between the minor alleles of eight measured SNPs (rs3020364, rs2474148, rs12154178, rs2347867, rs6927072, rs2982712, rs3020407, rs9322335) and percent mammographic density (p-values: 0.002-0.026), these associations were strongest in lean women (BMI, ≤23.6 kg/m2.). The odds of above-median percent mammographic density (>28.5 %) among women with major homozygous genotypes were 3-6 times higher than those of women with minor homozygous genotypes in seven SNPs. Women with rs3020364 major homozygous genotype had an OR of 6.46 for above-median percent mammographic density (OR: 6.46; 95 % Confidence Interval 1.61, 25.94) when compared to women with the minor homozygous genotype. These associations were not observed in relation to absolute mammographic density. No associations between SNPs and daily cycling estradiol were observed. However, we suggest, based on results of borderline significance (p values: 0.025-0.079) that the level of 17ß-estradiol for women with the minor genotype for rs3020364, rs24744148 and rs2982712 were lower throughout the cycle in women with low (<28.5 %) percent mammographic density and higher in women with high (>28.5 %) percent mammographic density, when compared to women with the major genotype. CONCLUSION: Our results support an association between eight selected SNPs in the ESR1 gene and percent mammographic density. The results need to be confirmed in larger studies.

A factor analysis approach to examining relationships among ovarian steroid concentrations, gonadotrophin concentrations and menstrual cycle length characteristics in healthy, cycling women.

STUDY QUESTION: How are ovarian steroid concentrations, gonadotrophins and menstrual cycle characteristics inter-related within normal menstrual cycles? SUMMARY ANSWER: Within cycles, measures of estradiol production are highly related to one another, as are measures of progesterone production; however, the two hormones also show some independence from one another, and measures of cycle length and gonadotrophin concentrations show even greater independence, indicating minimal integration within cycles. WHAT IS KNOWN ALREADY: The menstrual cycle is typically conceptualized as a cohesive unit, with hormone levels, follicular development and ovulation all closely inter-related within a single cycle. Empirical support for this idea is limited, however, and to our knowledge, no analysis has examined the relationships among all of these components simultaneously. STUDY DESIGN, SIZE, DURATION: A total of 206 healthy, cycling Norwegian women participated in a prospective cohort study (EBBA-I) over the duration of a single menstrual cycle. Of these, 192 contributed hormonal and cycle data to the current analysis. PARTICIPANTS/MATERIALS, SETTING, METHODS: Subjects provided daily saliva samples throughout the menstrual cycle from which estradiol and progesterone concentrations were measured. FSH and LH concentrations were measured in serum samples from three points in the same menstrual cycle and cycle length characteristics were calculated based on hormonal data and menstrual records. A factor analysis was conducted to examine the underlying relationships among 22 variables derived from the hormonal data and menstrual cycle characteristics. MAIN RESULTS AND THE ROLE OF CHANCE: Six rotated factors emerged, explaining 80% of the variance in the data. Of these, factors representing estradiol and progesterone concentrations accounted for 37 and 13% of the variance, respectively. There was some association between measures of estradiol and progesterone production within cycles; however, cycle length characteristics and gonadotrophin concentrations showed little association with any measure of ovarian hormone concentrations. LIMITATIONS, REASONS FOR CAUTION: Our summary measures of ovarian hormones may be imprecise in women with extremely long or short cycles, which could affect the patterns emerging in the factor analysis. Given that we only had data from one cycle on each woman, we cannot address how cycle characteristics may covary within individual women across multiple cycles. WIDER IMPLICATIONS OF THE FINDINGS: Our findings are generalizable to other healthy populations with typical cycles, however, may not be applicable to cycles that are anovulatory, extreme in length or otherwise atypical. The results support previous findings that measures of estradiol production are highly correlated across the cycle, as are measures of progesterone production. Estradiol and progesterone concentrations are associated with one another, furthermore. However factor analysis also revealed more complex underlying patterns in the menstrual cycle, highlighting the fact that gonadotrophin concentrations and cycle length characteristics are virtually independent of ovarian hormones. These results suggest that despite integration of follicular and luteal ovarian steroid production across the cycle, cycle quality is a multi-faceted construct, rather than a single dimension. STUDY FUNDING/COMPETING INTEREST(S): The EBBA-I study was supported by a grant from the Norwegian Cancer Society (49 258, 05087); Foundation for the Norwegian Health and Rehabilitation Organizations (59010-2000/2001/2002); Aakre Foundation (5695-2000, 5754-2002) and Health Region East. The current analyses were completed under funding from the National Institutes of Health (K12 ES019852). No competing interests declared.

Higher luteal progesterone is associated with low levels of premenstrual aggressive behavior and fatigue.

Contradictory findings show both positive and negative effect of progesterone on the premenstrual mood changes in women. Here we present the study investigating this relationship on the large sample of premenstrual women. 122 healthy, reproductive age women collected daily morning saliva samples and recorded intensity scores for the mood symptoms: irritability, anger, sadness, tearfulness, insomnia, and fatigue. Saliva samples were assayed for progesterone concentrations and mood intensity scores were used to calculate behavioral indices. Women with low Aggression/Irritability and Fatigue had consistently higher progesterone levels during the luteal phase than women with high Aggression/Irritability and Fatigue. Additionally, Aggression/Irritability and Fatigue correlated negatively with maximal progesterone value during the luteal phase. Our results demonstrated a negative effect of low progesterone level on the premenstrual mood symptoms such as aggressive behavior and fatigue in healthy reproductive age women. This supports a previously proposed model of biphasic action of progesterone metabolites on mood.

Ovarian hormones and reproductive risk factors for breast cancer in premenopausal women: the Norwegian EBBA-I study.

BACKGROUND: Ovarian hormones, parity and length of 'menarche-to-first birth' time interval are known risk factors for breast cancer, yet the associations between 17ß-estradiol, progesterone and these reproductive factors remain unclear. METHODS: A total of 204 women (25-35 years) who participated in the Norwegian EBBA-I study collected daily saliva samples for one complete menstrual cycle, and filled in a reproductive history questionnaire. Anthropometry was measured and saliva samples were analyzed for ovarian hormones. Associations between parity, the interval and ovarian hormones, and effects of hormone-related lifestyle factors were studied in linear regression models. RESULTS: Mean age was 30.7 years, and age of menarche 13.1 years. Parous women had on average 1.9 births, and age at first birth was 24.5 years. No association was observed between parity and ovarian steroids. In nulliparous women, higher waist circumference (≥ 77.75 cm) and longer oral contraceptive (OC) use (≥ 3 years) were associated with higher levels of 17ß-estradiol. Short (<10 years) versus long (>13.5 years) 'menarche-to-first birth' interval was associated with higher overall mean (P(trend) = 0.029), 47% higher maximum peak and 30% higher mid-cycle levels of 17ß-estradiol. We observed a 2.6% decrease in overall mean salivary 17ß-estradiol with each 1-year increase in the interval. CONCLUSIONS: Nulliparous women may be more susceptible to lifestyle factors, abdominal overweight and past OC use, influencing metabolic and hormonal profiles and thus breast cancer risk. Short time between 'menarche-to-first birth' is linked to higher ovarian hormone levels among regularly cycling women, suggesting that timing of first birth is related to fecundity.

Body fat, energy balance and estradiol levels: a study based on hormonal profiles from complete menstrual cycles.

BACKGROUND: Female fecundity is regulated by nutritional status. Although widely cited, this hypothesis is not strongly supported by empirical data from non-obese, healthy women of reproductive age. METHODS: Healthy, reproductive aged women (n = 141) from Southern Poland collected daily morning saliva samples for one complete menstrual cycle. Levels of 17-beta-estradiol were analyzed by radioimmunoassay. Anthropometric measurements, including body fat percentage, were taken randomly with respect to phase of the menstrual cycle. Energy balance was specified based on changes in body fat percentage from the beginning to the end of the observation period. RESULTS: Women with very low and high body fat had significantly lower levels of E2 compared with women with low and average body fat. In women of very low to average body fat, a 10% increase in body fat was associated with a 5-7 pmol/l increase in estradiol levels. The association between fat percentage and E2 was even stronger in women with positive energy balance, who also showed significant differences between body fat groups in estradiol profiles across whole the menstrual cycle. No such relationship was found in women with negative energy balance. CONCLUSIONS: In healthy women, we found a non-linear association between body fat and estradiol levels. Both very low and high body fat was associated with decreased estradiol levels. The relationship between estradiol and body fat was strongly influenced by women's energy balance.

17-beta-estradiol in relation to age at menarche and adult obesity in premenopausal women.

BACKGROUND: We hypothesize that premenopausal endogenous estradiol may be associated with age at menarche and adult overweight and obesity, potentially contributing to breast cancer risk. METHODS: We assessed age at menarche by questionnaire among 204 healthy Norwegian women, aged 25-35 years. Measures of body composition included body mass index (BMI, kg/m(2)), waist circumference (WC, cm), waist-to-hip ratio (WHR) and fat percentage dual energy X-ray absorptiometry, (DEXA). Daily salivary 17-beta-estradiol (E(2)) concentrations were collected throughout one entire menstrual cycle and assessed by radioimmunoassay (RIA). Linear regression analyses and linear mixed models for repeated measures were used and potential confounding factors and effect modifiers were tested. RESULTS: Among women with an early age at menarche (< or =12 years), the overall mean salivary E(2) concentration increased by 3.7 pmol/l (95% confidence interval, 1.8-5.7 pmol/l) with each 9.8 cm (1 SD) increase in WC, which represents a 20.7% change in the mean for the total group. Among the same early maturers, a 1 SD (0.06) change in WHR was directly associated with a 24.0% change in mean E(2) concentration for the total group. CONCLUSIONS: Our findings support the hypothesis that early age at menarche, together with adult overweight and obesity, result in high levels of 17-beta-estradiol throughout the menstrual cycle.

Men in committed, romantic relationships have lower testosterone.

Variation in human male testosterone levels may reflect, and effect, differential behavioral allocation to mating and parenting effort. This proposition leads to the hypothesis that, among North American men, those involved in committed, romantic relationships will have lower testosterone levels than men not involved in such relationships. Our study is the first to examine whether being in such a relationship (rather than being married) is the meaningful predictor of male testosterone levels. To test this hypothesis, 122 male Harvard Business School students filled out a questionnaire and collected one saliva sample (from which testosterone level was measured). Results revealed that men in committed, romantic relationships had 21% lower testosterone levels than men not involved in such relationships. Furthermore, the testosterone levels of married men and unmarried men who were involved in committed, romantic relationships did not differ, suggesting that, at least for this sample, male pair bonding status is the more significant predictor of testosterone levels than is marital status.

Energetic factors, ovarian steroids and the risk of breast cancer.

In industrial countries, women often have excess metabolic energy due to high food consumption and low physical activity. High lifetime energy availability results in high lifetime levels of ovarian steroid hormones. Oestrogens and progesterone are hypothesized to play a crucial role in the development and prognosis of breast cancer. Epidemiological studies document the importance of physical activity and caloric limitations in reducing breast cancer risk. The risk of breast cancer is much higher in industrial countries than in developing countries, where women are characterized by lower energy intake and higher energy expenditure. It is likely, that the beneficial effects of physical activity and of negative energy balance are mediated by the reduced levels of ovarian steroids. While both weight loss and physical activity may have similar efficacy in suppressing ovarian function and, therefore, in reducing the risk of breast cancer, we suggest that it may be more advantageous for premenstrual women to achieve lifetime reduction in steroid levels by increasing their physical activity, rather than by weight loss due to caloric restriction alone.

Does hypoxia impair ovarian function in Bolivian women indigenous to high altitude?

Fertility appears to be reduced in at least some high altitude populations relative to their counterparts at lower elevations. Inferring from the difficulties with reproduction of newcomers to high altitude and from animal experiments, it has been hypothesized that this apparent reduction is the result of hypoxia acting to reduce fecundity and/or increase fetal loss. In humans, however, several behavioral as well as biological factors may affect fertility levels. These many factors have been organized by demographers into a framework of seven proximate determinants that includes fecundability (the monthly probability of conception) of which successful ovulation is one component. To test whether ovarian function is impaired in women indigenous to high altitude, we measured salivary progesterone (P) in a sample (n = 20) of Quechua women (aged 19-42 years) residing at 3,100 m. It was found that mean luteal P = 179 pmol/L and mean midluteal P = 243 pmol/L, levels that fall about midway in the range of known values for several populations and are higher than some lower altitude populations. These findings suggest that hypoxia does not appear to significantly impair ovarian function in those with lifelong residence at high altitude. There are, however, several factors common to many high altitude populations that may act to reduce fecundability and fertility including intercourse patterns (affected by marriage and migration practices), prolonged lactation, dietary insufficiency, and hard labor.

Women's strategies to alleviate nutritional stress in a rural African society.

Many agricultural populations are subject to chronic or seasonal undernutrition, reproductive women and children often being most vulnerable. This paper presents quantitative and qualitative data on food consumption, food distribution practices, food taboos, garden sizes and work effort to show how Lese horticulturalist women living in the Ituri Forest of northeast Democratic Republic of Congo attempt to alleviate nutritional stress. The Lese experience an annual hunger season when approximately one quarter of the population suffer from energy deficiency. Nutritional intake is also compromised by a complex system of food taboos against meat from wild forest animals. Anthropometric data collected over several years suggest that Lese women suffer from nutritional stress more than men during the hunger season. They also have more food taboos particularly during pregnancy and lactation. Their low fertility is compounded by nutritional stress. Despite these inequities, Lese women use several strategies to improve their food intake. Since they are responsible for all household cooking, they manipulate food portions. During the hunger season, they snack frequently, and increase their consumption of palliative foods. Women with more food taboos plant larger gardens to supplement their diet with vegetable foods. Although this results in their consumption of more daily protein, they work harder compared to women with smaller gardens. Women cheat in their adherence to specific food taboos by actively discounting them, or by eating prophylactic plants that supposedly prevent the consequences (usually illness) of eating tabooed foods. In addition, women resort to subterfuge to access desirable resources. Lese women do not reduce work effort during the hunger season, but adapt physiologically by reducing resting metabolic rates during periods of weight loss. These results point to the ability of Lese women to minimize the ecological and cultural constraints on their nutrition. More data, however, are required to assess the long-term effectiveness of these strategies.

Physical work causes suppression of ovarian function in women.

The suppression of reproductive function is known to occur in women engaging in activities that require high energetic expenses, such as sport participation and subsistence work. It is still unclear, however, if reproductive suppression is a response to high levels of energy expenditure, or only to the resulting state of negative energy balance. To our knowledge, this study provides the first evidence that work-related energy expenditure alone, without associated negative energy balance, can lead to the suppression of reproductive function in women. We document suppression of ovarian function expressed as lowered salivary progesterone levels in women from an agricultural community who work hard, but remain in neutral energy balance. We propose two alternative evolutionary explanations (the 'pre-emptive ovarian suppression' hypothesis and the 'constrained down-regulation' hypothesis) for the observed results.

Dietary composition and ovarian function among Lese horticulturalist women of the Ituri Forest, Democratic Republic of Congo.

OBJECTIVE: To assess dietary composition in relation to energetic status and ovarian function. DESIGN: An eight-month prospective study undertaken in 1989. SETTING: Ituri Forest, northeast Democratic Republic of Congo. SUBJECTS: Sixty-four female volunteers of reproductive age (16-45), with a subsample of 30 for nutritional focal follows, 39 for analysis of salivary progesterone levels, and 18 for analysis of salivary oestradiol. INTERVENTIONS: Regular anthropometric measurements of heights, weights, and mid-triceps skinfolds, 204 h of nutritional focal follows when all foods prepared and consumed were recorded and weighed, analysis of dietary composition using African food tables, collection of saliva samples every-other day for radioimmunoassay of salivary oestradiol and progesterone. RESULTS: The primary staple in the Lese diet is manioc, consumed at nearly every meal, but the diet is subject to seasonal fluctuations. Important seasonal crops are groundnuts and rice. Carbohydrates comprise 64% of the diet, protein 14% and fat 22%, but seasonal fluctuations in food items create significant differences in protein content (P = 0.007). Energy intake falls by 25% in the hunger season. Lese energy balance reflects this lack with women losing a mean 7% of total body weight. The mean BMI for Lese women at the beginning of the study was 22.8 but, during the hunger season, the BMI for 20% of women fell to < 18.5, indicative of chronic energy shortage. Mean levels of Lese salivary oestradiol and progesterone are chronically and significantly lower than healthy, Western controls (P = 0.0001 for progesterone; P = 0.03 for follicular values of oestradiol, P = 0.0001 for mid-cycle values, and P = 0.0002 for luteal values). Mean salivary progesterone levels were significantly lower for those Lese women losing weight (P = 0.03), and significantly lower for weight-loss women when comparing levels at the beginning and end of the hunger season (P = 0.03). DISCUSSION: The relatively low-fat, high-fibre diet of the Lese appears healthy, but there are occasional seasonal deficiencies in protein-energy and micronutrient content, especially for pregnant and lactating women. Dietary composition may affect Lese levels of reproductive steroids, partly explaining the chronically low salivary oestradiol profile of this population.

Salivary testosterone levels among Tamang and Kami males of central Nepal.

Salivary testosterone levels are reported for 65 Nepalese males between the ages of 15 and 48 years who were drawn from 2 different ethnic populations (Tamang and Kami) from the central highlands of Nepal. Subjects collected morning and evening saliva samples on five consecutive days in two contrasting seasons, the winter dry season and the summer monsoon season. Anthropometric indexes of acute and chronic nutritional status were also measured. Morning and evening salivary testosterone levels in the winter averaged 233 +/- 14 (SE) pmol/L and 166 +/- 8 pmol/L, respectively, for the Tamang and 249 +/- 14 pmol/L and 163 +/- 13 pmol/L, respectively, for the Kami. In the summer the corresponding values were 219 +/- 12 pmol/L and 156 +/- 8 pmol/L for the Tamang and 249 +/- 19 pmol/L and 147 +/- 12 pmol/L for the Kami. These levels are significantly lower than those reported for Western populations and close to those reported for other non-Western populations. The magnitude of diurnal variation in salivary testosterone levels and the absence of significant age variation are also comparable with observations made on other populations. Weak relationships were observed between testosterone levels and indexes of acute and chronic nutritional status in the winter only. The absence of pronounced variation in salivary testosterone levels between populations and the absence of strong associations between salivary testosterone levels and indexes of acute and chronic nutritional status contrast with the prominent ecological and interpopulation variation reported for salivary progesterone levels in women. Male gonadal function seems less sensitive to moderate energetic stress than female gonadal function, probably reflecting the fact that energy availability is less crucial to male reproductive success than to female reproductive success. Variation in testosterone level associated with chronic energetic stress may be an adaptive somatic response to avoid the maintenance costs of a large active metabolic mass with little direct impact on male fecundity.

Comparison of salivary steroid profiles in naturally occurring conception and non-conception cycles.

Oestradiol and progesterone profiles from naturally occurring conception and exposed non-conception cycles were compared to assess the impact of natural variation in concentrations of ovarian steroid hormones on female fecundity. In a prospective, longitudinal study, 24 women collected saliva samples twice daily and recorded intercourse for up to 1 year or until a pregnancy was clinically confirmed. Oestradiol and progesterone concentrations were measured by a salivary radioimmunoassay. Average mid-follicular oestradiol concentrations were significantly higher in conception than in non-conception cycles (12.6 +/- 1.7 versus 8.5 +/- 0.6 pmol/l, P < 0.01). A separate analysis, including only cycles from those women who contributed both conception and non-conception cycles, demonstrated an even more pronounced difference in mid-follicular oestradiol concentrations, not just for conception and non-conception cycles as groups (14.5 +/- 2.3 versus 6.5 +/- 0.7 pmol/l, P < 0.001), but also between the conception and average non-conception concentrations of individual women. Among these women, relative mid-follicular oestradiol concentration was highly correlated with the probability of successful conception. In addition, relative body weight was significantly positively correlated with mid-follicular oestradiol concentration. These findings indicate that variation in follicular development, reflected in variation in follicular oestradiol concentrations, is an important indicator of fecundity.

Reference values for luteal progesterone measured by salivary radioimmunoassay.

OBJECTIVE: To establish age-stratified reference values for salivary luteal P levels. DESIGN: One hundred thirty-six regularly menstruating women (18 to 48 years of age), screened for weight, exercise, and steroid medication use, collected daily saliva samples for one complete menstrual cycle. Luteal P levels were measured by 3H-RIA, and data were aligned by day of next menstrual onset. Means (+/- 1 SD range) and percentiles, calculated using both untransformed and log transformed data, were calculated for each luteal day and for indices of luteal P production. RESULTS: Reference values for salivary daily luteal P levels and indices of luteal P are presented for three age groups (18 to 24 years, 25 to 39 years, and 40 to 48 years). CONCLUSION: The age-stratified reference values presented here can be used, without collateral clinical procedures, to assess salivary luteal P levels. Salivary monitoring is ideally suited for research and long-term clinical observation, but the characteristics of salivary P data may limit the usefulness of these values for individual diagnosis.

Advances in human reproductive ecology.

PIP: Human reproductive ecology pertains to reproduction biology and changes due to environmental influences. The research literature relies on clinical, epidemiological, and demographic analysis. The emphasis is on normal, nonpathological states and a broad range of ecological conditions. This review focused on the importance of age and energetic stress from ecological conditions rather than dieting or self-directed exercise in changing female fecundity. The literature on male reproductive ecology is still small but growing. J.W. Wood provided a comprehensive overview of the field. Natural fertility, as defined by Henry, is the lack of parity-specific fertility limitation. There is evidence that fertility can vary widely in natural fertility populations. There are consistent age patterns among different natural fertility populations. Doring found that there was higher frequency of anovulatory and luteal insufficiency in cycles during perimenarche and perimenopausal periods. Infertility studies have shown declines in pregnancy rates in women over the age of 30 years. Ovum donation evaluations have found both uterine age and ovarian and oocyte age to be related to the probability of a successful pregnancy. Basal follicle stimulating hormone and the endometrial thickness are important predictors of ovarian capacity and related to age and declining fecundity. Much of the literature on fecundity is derived from women with impaired reproductive physiology. In Lipson and Ellison's study of healthy women, average follicular and average luteal estradiol values declined with increasing subject age. Low follicular levels were correlated with smaller follicular size, low oocyte fertilizability, reduced endometrial thickness, and low pregnancy rates. Comparisons across populations have shown that populations experience declines in luteal function with age, but levels of luteal functions varied widely. Chronic conditions which slow growth and delay reproductive maturation may impact on lower ovarian function throughout adult life. There is a range of ovarian function along a continuum due to energetic stress. Evidence from the Lese in Zaire, the Tamang of Nepal, and Polish farm women outside Crakow suggest that workload affects ovarian function. Luteal function and ovulatory frequency is lower when women are losing weight. Among the Tamang losing weight between seasons there was evidence of lower ovarian function during the monsoon season. Polish farm women who work very hard in summer had lower ovarian function. The effect of lactation on amenorrhea appears to be due to the energetic stress on the mother in the intensity and duration of suckling. Women in poorer nutritional status may require more intense suckling. Seasonality of energy balance may be related to seasonality of female fecundity and conceptions.^ieng

Extinction and descent.

The probability of lineal extinction is sensitive to all the moments of the reproductive success probability distribution. In particular, high variance in reproductive success is associated with high probability of lineal extinction. Where male variance in reproductive success exceeds female variance, strictly patrilineal lines of descent will become extinct more rapidly than strictly matrilineal lines of descent. Patrilineal genealogies will be expected to be shallower and broader than matrilineal genealogies under such conditions. Potential implications of this genealogical asymmetry for human descent systems include the greater information content of patrilineal kinship reckoning compared with any other unilineal system and the greater effectiveness of patrilineal kinship as a vehicle for corporate action.