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Among adults, aldosterone and cortisone increases are reported in response to physically taxing forms of competition, enabling individuals to rapidly adapt to variable sociocompetitive contexts. Yet, aldosterone and cortisone responses in juvenile children engaged in less strenuous forms of competition have not been investigated. Here, we sought to measure aldosterone and cortisone responses in children who participated in math and table-tennis competitions. We hypothesized that the responses would significantly vary with respect to the type of competition. Pre- and post-match salivary aldosterone and cortisone were measured in Hong Kongese children, aged 8–11 years, during (1) a mixed-sex, team-based, math competition (N = 45) and (2) a dyadic, table-tennis competition against peers (N = 22). In the math competition, aldosterone and cortisone levels decreased in boys and girls, while members on losing teams had greater match decreases in cortisone levels compared to individuals on winning teams. In the table-tennis competition, time of day led to significant diurnal differences in competitors’ pre-match aldosterone and cortisone concentrations. As a result, each sample was analyzed independently according to match time (8:30 AM and 11:00 AM). Aldosterone levels decreased among the competitors who participated in the 11:00 AM table-tennis matches. Cortisone levels decreased for the majority of competitors, but only significantly decreased in the 8:30 AM sample. These findings highlight that juvenile competitors’ hypothalamic-pituitary-adrenal (HPA) axis and renin-angiotensin-aldosterone system (RAAS) are sensitive to less physically strenuous forms of competition. Further, the differences in the competitive environment likely stimulate the direction of aldosterone (RAAS) and cortisone (HPA) reactive change.

Introduction Hormones are the chemical messengers of your body. Steroid hormones like testosterone are a specific class of hormone that are produced predominantly in the gonads (sex steroids) and less so in the adrenal glands (corticosteroids). When too little or too much of testosterone is produced, adverse symptoms and conditions can occur. In order to restore the vital balance of hormones, detailed and accurate measurement of hormones is needed. Serum currently represents the gold standard when it comes to hormone testing. However, there are other body fluids such as saliva, urine, and capillary blood (finger prick) where the hormone levels correlate well with serum. A significant advantage of these alternative fluids is that they are non-invasive, or semi-invasive, can be collected "on the go" by the patient at home, and don't have the strict requirements for cold-chain shipping. Objectives The primary objective of this work is to show that finger-prick dried blood spot blood is quantitatively equivalent to serum when compared by LC-MS/MS testing in patients only producing endogenous testosterone. We will also show that topically delivered testosterone increases in a dose-dependent fashion, whereas serum levels of testosterone increase very little regardless of topical testosterone dosing. Methods Serum samples were collected by a physician and sent into the lab and analyzed for testosterone on a Siemens Immulite 2000 autoanalyzer. Dried blood spots were collected by the patient and sent into the lab and analyzed on an AB Sciex 5500 triple quadrupole mass spectrometer. Results Patients who don't supplement with topical testosterone show a strong correlation between endogenous venipuncture serum and DBS testosterone levels. In sharp contrast, samples from topical testosterone users show remarkably different results with hormone levels 10-100×higher in the DBS than serum results. Conclusion These results are significant because it gives a glimpse into which forms of testing are most appropriate for different patients based on the mode of steroid hormone delivery. Without using the proper body fluid for testosterone detection, patients potentially could be overdosed if serum results are used to guide therapeutic dosing. Also, venipuncture serum will fail to show "sports-doping" with exogenous testosterone, which easily can be identified with capillary finger stick blood. This is also significant as the World Anti-doping Agency has finally begun accepting these data as fact and have listed dried blood spot as an appropriate form of collection and analysis for drug testing. This can help find athletes using performance enhancers that would normally have skirted by the system (1). WADA. (2021, May 21) WADA Executive Committee approves Technical Document for innovative dried blood spot testing method. https://www.wada-ama.org/en/media/news/wada-executive-committee-approves-technical-document-for-innovative-dried-blood-spot. Presentation: Sunday, June 12, 2022 12:36 p.m. - 12:41 p.m., Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Background The International Cohort on Lifestyle Determinants of Health (INCLD Health) is an ongoing, prospective cohort study assessing the health behaviours and lifestyles of higher education students, including their use of specialty diets and complementary and integrative health (CIH) practices. Purpose: This cross-sectional analysis of the INCLD Health cohort aims to (1) evaluate the associations between perceived stress, sleep disturbance, and meditative practices with diurnal salivary free cortisol and (2) evaluate the associations of meditative practices as well as mind-body practices with perceived stress and sleep disturbance. Methods Serial multivariable linear regression models, adjusting for sociodemographic and lifestyle behaviours, were used to assess associations of (1) perceived stress, sleep disturbance, and meditative practices with salivary cortisol, and (2) meditative practices as well as mind-body practices with perceived stress and sleep disturbance. Meditative and mind-body practices were evaluated using a stress-management and self-care survey; perceived stress and sleep disturbance were evaluated using the 10-item Perceived Stress Scale (PSS), and the patient reported outcome measures information system-29 (PROMIS-29) sleep sub-score respectively. Salivary cortisol was collected at 4 time points over a 24-hour period and area under the curve (AUC) calculations conducted. Results 82.5% (n = 80) of participants utilized at least monthly meditative practices. Greater disturbed sleep, but not perceived-stress, meditative, nor mind-body practices was independently associated with increased AUC cortisol (b = 0.02, 95% CI: 0.002–0.05, p = 0.03) after adjusting for age, sex, race, ethnicity, and BMI. Neither meditative nor mind-body practices were associated with perceived stress or disturbed sleep. Conclusions Among INCLD Health participants, greater sleep disturbance, but not perceived stress or meditative practices were associated with daytime cortisol.

STUDY QUESTION Is psychosocial stress associated with ovarian function in reproductive-aged survivors of cancer diagnosed as adolescents and young adults (AYA survivors)? SUMMARY ANSWER We observed no association between self-reported and biomarkers of psychosocial stress and ovarian function in AYA survivors. WHAT IS KNOWN ALREADY Psychosocial stress suppresses hypothalamic-pituitary-ovarian axis, resulting in ovulatory dysfunction, decreased sex steroidogenesis and lower fertility in reproductive-aged women. Many cancer survivors experience high psychosocial stress and hypothalamic-pituitary-adrenal axis dysregulation. The menstrual pattern disturbances and infertility they experience have been attributed to ovarian follicle destruction, but the contribution of psychosocial stress to these phenotypes is unknown. STUDY DESIGN, SIZE, DURATION A cross-sectional study was conducted estimating the association between perceived stress, measured by self-report and saliva cortisol, and ovarian function, measured by bleeding pattern, dried blood spot (DBS) FSH and LH, and saliva estradiol. We included 377 AYA survivor participants. PARTICIPANTS/MATERIALS, SETTING, METHODS AYA survivor participants were ages 15–35 at cancer diagnosis and ages 18–40 at study enrollment, had completed primary cancer treatment, had a uterus and at least one ovary, did not have uncontrolled endocrinopathy and were not on hormone therapy. Recruited from cancer registries, physician referrals and cancer advocacy groups, participants provided self-reported information on psychosocial stress (Perceived Stress Scale-10 (PSS-10)) and on cancer and reproductive (fertility, contraception, menstrual pattern) characteristics. DBS samples were collected timed to the early follicular phase (cycle Days 3–7) for menstruating individuals and on a random day for amenorrheic individuals; saliva samples were collected three time points within 1 day. FSH and LH were measured by DBS ELISAs, cortisol was measured by ELISA and estradiol was measured by liquid chromatography tandem mass spectrometry. MAIN RESULTS AND THE ROLE OF CHANCE The median age of participants was 34.0 years (range 19–41) at a median of 6.0 years since cancer diagnosis. The most common cancer was breast (32.1%). Median PSS-10 score was 15 (range 0–36), with 5.3% scoring ≥26, the cut point suggestive of severe stress. Cortisol levels followed a diurnal pattern and cortisol AUC was negatively correlated with PSS-10 scores (P = 0.03). Neither PSS-10 scores nor cortisol AUC were associated with FSH, LH, estradiol levels or menstrual pattern. Waking and evening cortisol and the cortisol awakening response also were not related to ovarian function measures. LIMITATIONS, REASONS FOR CAUTION Our analysis is limited by its cross-sectional nature, heterogeneity of cancer diagnosis and treatments and low prevalence of severe stress. WIDER IMPLICATIONS OF THE FINDINGS The lack of association between psychosocial stress and a variety of ovarian function measures in female AYA cancer survivors suggests that psychosocial stress does not have a significant impact on the reproductive axis of AYA survivors. This finding is important in counseling this population on their menstrual pattern and family building plans. STUDY FUNDING/COMPETING INTEREST(S) NIH HD080952, South Korea Health Industry Development Institute HI18C1837 (JK). Dr A.D. works for Bluebird Bio, Inc., Dr D.Z. works for ZRT Labs and Dr P.M.S. works for Ansh Labs, which did not sponsor, support or have oversight of this research. Other authors report no competing interests. TRIAL REGISTRATION NUMBER N/A

Aldosterone and cortisone are released in response to physical and psychological stress. However, aldosterone and cortisone responses in children engaged in physical competition have not been described. We examined salivary aldosterone and salivary cortisone responses among Hong Kongese boys, aged 8-11 years, during (1) a soccer match against unknown competitors (

Knight, McShane, et al. (2020) report three experiments on testosterone’s effect on the Cognitive Reflection Test (CRT). The experiments were designed and executed independently of each other and of our previous work (Nave, Nadler, Zava & Camerer, 2017). We thank Knight, McShane, et al. for conducting these experiments and summarizing their results, and we agree that one experiment is obviously not enough for establishing an empirical fact. The individual experiments and their meta-analytic summary are consistent with both the null hypothesis and Nave et al.’s conclusions (see Table S6 in Knight, McShane, et al.’s Supplemental Material), and there is evidence for variation in effects across experiments. In what follows, we reflect on design differences among the experiments and the collective evidence that their data contain.

Context During puberty, LH pulse frequency increases during sleep; in women, LH pulse frequency slows during sleep in the early/middle follicular phase (FP) of the menstrual cycle. The origin and significance of this developmental transition are unknown. Objective To determine the relationship between progesterone (P4) exposure, sleep-related slowing of LH pulses in the FP, and the intercycle FSH rise, which promotes folliculogenesis, in early postmenarchal girls. Methods 23 girls (gynecologic age 0.4 to 3.5 years) underwent hormone measurements and pelvic ultrasounds during two consecutive cycles and one frequent blood sampling study with concurrent polysomnography during the FP. Results Subjects demonstrated one of four patterns during cycle 1 that represent a continuum of P4 exposure: ovulatory cycles with normal or short luteal phase lengths or anovulatory cycles ± follicle luteinization. Peak serum P4 and urine pregnanediol (Pd) in cycle 1 were inversely correlated with LH pulse frequency during sleep in the FP of cycle 2 (r = -0.5; P = 0.02 for both). The intercycle FSH rise and folliculogenesis in cycle 2 were maintained after anovulatory cycles without P4 or Pd exposure or nocturnal slowing of LH pulse frequency in the FP. Conclusions During late puberty, rising P4 levels from follicle luteinization and ovulation may promote a slower LH pulse frequency during sleep in the FP. However, a normal FSH rise and follicle growth can occur in the absence of P4-associated slowing. These studies therefore suggest that an immature LH secretory pattern during sleep is unlikely to contribute to menstrual irregularity in the early postmenarchal years.

Context Menstrual irregularity after menarche has been attributed to immature estrogen positive feedback activity (E+FB) but data are conflicting. Objective To determine the hypothalamic-pituitary-ovarian contributions to menstrual irregularity in adolescents. Methods Twenty-three healthy girls [aged 12.8 to 17.6 years; 0.4 to 3.5 years postmenarche; body mass index (BMI) percentile, 41.0 to 99.3] underwent serial hormone measurements and pelvic ultrasounds during two consecutive menstrual cycles. Hormones and follicle growth were compared with 65 adult historic controls with ovulatory cycles (OVs). Results Girls had anovulatory cycles (ANOVs; 30%), OVs with a short luteal phase (short OVs; 22%), or OVs with normal luteal phase (normal OVs; 48%) without differences in cycle length, chronologic or gynecologic age, or BMI. Adolescents showed a spectrum of E+FB [midcycle LH adjusted for preovulatory estradiol (E2)]; only normal OV girls were comparable to adults. All OV girls had lower E2, progesterone, and gonadotropins during the luteal phase and luteal-follicular transition compared with adults. Normal OV girls also had lower follicular phase LH and FSH levels, a longer follicular phase, a slower dominant follicle growth rate, and smaller estimated preovulatory follicle size than adults. Follicular phase E2 and inhibin B levels were lower in normal OV girls than in adults even after adjusting for differences in FSH and follicle size. Conclusions Early postmenarchal girls with normal OVs demonstrate mature E+FB but continue to have lower gonadotropin levels, diminished ovarian responsiveness, and decreased corpus luteum sex steroid synthesis compared with adults, indicating that reproductive axis maturity requires coordinated development of all components of the hypothalamic-pituitary-ovarian axis.

Esports, or competitive video gaming, has rapidly increased in online play and viewing. The popularity of esports such as League of Legends may derive in part because it features skills-based coalitional competition. Whereas a sizable literature focuses on adult human hormones and competition, little research has addressed the hormone responses of men playing video games. The purpose of the present study is to investigate the effects of playing a coalitionary-based esport on young U.S. men's steroid hormone levels in a naturalistic study. We tested salivary steroid changes in response to esports club members (n = 26) playing League of Legends against other people and the computer. We hypothesized that esports competition would increase testosterone, cortisol, DHEA and androstenedione levels, with more pronounced increases in winners than losers. Participants provided saliva samples before and after competitions lasting 15–27 min in duration. Salivary testosterone, cortisol, DHEA and androstenedione levels did not change overall or between play against people vs. the computer or with respect to winning or losing. However, play duration (range 16–27 min) was positively related to changes in DHEA, androstenedione and testosterone during play against people. Aldosterone levels decreased overall. We suggest that the informal and familiar environment as well as relatively short play duration help account for generally null findings. These findings help document physiological effects of esport play, in turn contributing to a richer understanding of why so many play and watch esports.

Little psychoneuroendocrine research has focused on steroid hormone responses to non-physical competition in middle childhood. This study sought to observe testosterone, estradiol, dehydroepiandrosterone (DHEA), androstenedione, and cortisol responses in children during a mixed-sex, team, academic competition. Salivary steroid hormones were collected, along with measures of performance, Body Mass Index, and pubertal development in ethnically Chinese boys (n = 18) and girls (n = 27), aged 9–10 years, during a math competition (N = 45). Testosterone and estradiol levels were generally low and unmeasurable. Nearly every competitor experienced decreases in cortisol and cortisol/DHEA molar ratio. Pre- and post-match DHEA and androstenedione did not significantly change. Exploratory analyses revealed a positive correlation between DHEA change and team performance among non-active participants (i.e. did not attempt to answer a question). ANCOVAs revealed differences in percentage change in androstenedione between active (n = 20) and non-active participants (n = 25) and among winners (n = 7) and losers (n = 38), and positive associations with age. Percentage change in cortisol was significantly lower among losers compared to winners. Performance measures were positively correlated with DHEA change and percentage change in androstenedione. Despite girls having higher pre-match androstenedione, both sexes exhibited similar patterned hormone responses. Only cortisol and cortisol/DHEA molar ratio decreased during the competition. However, DHEA, androstenedione, and cortisol match changes were partially related to psychosocial variables (e.g., performance, outcome, participation). These findings provide new insight into factors which may underpin steroid hormone responses during middle childhood non-athletic competition.

In nonhumans, the sex steroid testosterone regulates reproductive behaviors such as fighting between males and mating. In humans, correlational studies have linked testosterone with aggression and disorders associated with poor impulse control, but the neuropsychological processes at work are poorly understood. Building on a dual-process framework, we propose a mechanism underlying testosterone’s behavioral effects in humans: reduction in cognitive reflection. In the largest study of behavioral effects of testosterone administration to date, 243 men received either testosterone or placebo and took the Cognitive Reflection Test (CRT), which estimates the capacity to override incorrect intuitive judgments with deliberate correct responses. Testosterone administration reduced CRT scores. The effect remained after we controlled for age, mood, math skills, whether participants believed they had received the placebo or testosterone, and the effects of 14 additional hormones, and it held for each of the CRT questions in isolation. Our findings suggest a mechanism underlying testosterone’s diverse effects on humans’ judgments and decision making and provide novel, clear, and testable predictions.

The capacity to infer others' mental states (known as ‘mind reading’ and ‘cognitive empathy’) is essential for social interactions across species, and its impairment characterizes psychopathological conditions such as autism spectrum disorder and schizophrenia. Previous studies reported that testosterone administration impaired cognitive empathy in healthy humans, and that a putative biomarker of prenatal testosterone exposure (finger digit ratios) moderated the effect. However, empirical support for the relationship has relied on small sample studies with mixed evidence. We investigate the reliability and generalizability of the relationship in two large-scale double-blind placebo-controlled experiments in young men ( n = 243 and n = 400), using two different testosterone administration protocols. We find no evidence that cognitive empathy is impaired by testosterone administration or associated with digit ratios. With an unprecedented combined sample size, these results counter current theories and previous high-profile reports, and demonstrate that previous investigations of this topic have been statistically underpowered.

In modern human cultures where social hierarchies are ubiquitous, people typically signal their hierarchical position through consumption of positional goods—goods that convey one’s social position, such as luxury products. Building on animal research and early correlational human studies linking the sex steroid hormone testosterone with hierarchical social interactions, we investigate the influence of testosterone on men’s preferences for positional goods. Using a placebo-controlled experiment (N = 243) to measure individuals’ desire for status brands and products, we find that administering testosterone increases men’s preference for status brands, compared to brands of similar perceived quality but lower perceived status. Furthermore, testosterone increases positive attitudes toward positional goods when they are described as status-enhancing, but not when they are described as power-enhancing or high in quality. Our results provide novel causal evidence for the biological roots of men’s preferences for status, bridging decades of animal behavioral studies with contemporary consumer research., Testosterone is believed to be involved in social rank-related behavior. Here, the authors show that one dose of testosterone increases men’s preference for “high status” goods and brands, suggesting a role for testosterone in modern consumer behavior in men.

A large body of research links testosterone and cortisol to male-male competition. Yet, little work has explored acute steroid hormone responses to coalitional, physical competition during middle childhood. Here, we investigate testosterone, dehydroepiandrosterone (DHEA), androstenedione, and cortisol release among ethnically Chinese boys in Hong Kong (N = 102), aged 8–11 years, during a soccer match (n = 84) and an intrasquad soccer scrimmage (n = 81), with 63 participants competing in both treatments. The soccer match and intrasquad soccer scrimmage represented out-group and in-group treatments, respectively. Results revealed that testosterone showed no measurable change. DHEA increased during both treatments in the majority of participants and the degree of change had no relation to independent variables (e.g., performance, age, treatment, outcome) or covariate measures (Body Mass Index, Pubertal Development Scale). Most boys experienced androstenedione increases during match play, but no significant differences during the intrasquad soccer scrimmage competitions. The magnitude of change differed significantly between treatments and was positively associated with age. These latter findings suggest boys’ androstenedione responses may be sensitive to competitor type (i.e., unknown competitors vs. peers). For most subjects, cortisol significantly increased during match play, decreased during the intrasquad soccer scrimmage, and differed significantly between treatments, suggesting each treatment promoted a different psychological state among competitors. Cortisol/DHEA molar ratio decreased during the intrasquad scrimmage, suggestive of a more relaxed mental state. These data shed new light on potential proximate mechanisms associated with coalitional competition among prepubescent boys, with relevance to adrenarche and life history theory.

We examined potential changes in salivary DHEA, androstenedione, cortisol, testosterone, and cortisol/DHEA ratio in boys in response to soccer practice and soccer match competition. To our knowledge, this study is the first to explore the impacts of athletic competition on salivary steroid hormone change in juvenile boy soccer players. Boys from five different Las Vegas, Nevada teams provided saliva samples during soccer practice (N = 28), with four out of five teams providing saliva samples during a soccer match (N = 26). All participants were aged 8 – 10 years. A Wilcoxon Signed-Rank Sum Test and standard linear regression analyses were performed to assess the change in hormone concentrations during soccer practice and soccer match competition. Results revealed a statistically significant rise in boys’ DHEA concentrations during both practice and match play. Androstenedione significantly increased during the match competition only. Cortisol did not change with statistical significance during either condition. A Friedman’s ANOVA was used to evaluate within-subject differences in boys who participated in both the practice and match treatments (N = 17). Cortisol was the only hormone measure that, with statistical significance, increased more during the match than it did during practice. Analysis of the cortisol/DHEA ratios revealed a statistically significant decrease occurred during practice only. No statistical analysis was available for testosterone since all but seven samples were below the sensitivity of the assay (

There have been few studies of whether vitamin D insufficiency is linked with depression in healthy young women despite women’s high rates of both problems. Female undergraduates (n = 185) living in the Pacific Northwest during fall, winter, and spring academic terms completed the Center for Epidemiologic Studies Depression (CES-D) scale weekly for four weeks (W1–W5). We measured serum levels of vitamin D3 and C (ascorbate; as a control variable) in blood samples collected at W1 and W5. Vitamin D insufficiency (

As first responders who are frequently exposed to job-related trauma, police officers are at an elevated risk of adverse mental and physical health outcomes. Evidence-based approaches to stress reduction are sorely needed to address the complex variety of problems that police officers face. In this pilot study we examined the feasibility and preliminary effectiveness of a mindfulness-based intervention designed to address police officer stress. A total of 43 police officers completed an 8-week Mindfulness-Based Resilience Training (MBRT) program, which was designed to improve mindfulness, resilience, stress, health outcomes, and emotional functioning. Using multilevel models we found significant improvement in self-reported mindfulness, resilience, police and perceived stress, burnout, emotional intelligence, difficulties with emotion regulation, mental health, physical health, anger, fatigue, and sleep disturbance. Although there were no significant pre-to-post-MBRT changes in cortisol awakening response (CAR), while controlling for pre-MBRT increase area under the curve (AUCI), change in mental health was a significant predictor of post-AUCI. Implications of these findings and areas for future research are discussed.

John A Sturgeon,1 Beth D Darnall,1 Heather L Zwickey,2 Lisa J Wood,3 Douglas A Hanes,2 David T Zava,4 Sean C Mackey1 1Stanford University School of Medicine, Department of Anesthesiology, Perioperative and Pain Medicine, Palo Alto, CA, USA; 2Helfgott Research Institute, National College of Natural Medicine, Portland, OR, USA; 3MGH Institute of Health Professions, Boston, MA, USA; 4ZRT Laboratories, Beaverton, OR, USA Abstract: Though fibromyalgia is not traditionally considered an inflammatory disorder, evidence for elevated inflammatory processes has been noted in this disorder in multiple studies. Support for inflammatory markers in fibromyalgia has been somewhat equivocal to date, potentially due to inattention to salient patient characteristics that may affect inflammation, such as psychiatric distress and aging milestones like menopause. The current study examined the relationships between proinflammatory cytokines and hormone levels, pain intensity, and psychological distress in a sample of 34 premenopausal and postmenopausal women with fibromyalgia. Our results indicated significant relationships between interleukin-8 and ratings of pain catastrophizing (r=0.555, P

OBJECTIVES Little is known about salivary steroid hormone responses to dyadic competition among prepubescent boys. The current study explored pre-match and post-match testosterone, dehydroepiandrosterone (DHEA), androstenedione, and cortisol among 22 ethnically Chinese, Hong Kongese table tennis athletes, aged 8-11 years, during dyadic competition against peers. These data provide novel comparative insight into boys' hormone responses when participating in similar forms of competition to that of adults. METHODS Measures of salivary steroid hormones, age, outcome, and participant's self-reported perceived performance were obtained. Pre-match salivary steroid hormones and competition-induced steroid hormone changes were explored to further assess overall hypothalamic-pituitary-adrenal axis activity. RESULTS Cortisol decreased for most participants, whereas testosterone measures were below the sensitivity of the assay. DHEA and androstenedione did not significantly change during the table tennis exhibitions and were unrelated to independent performance variables. Correlational analyses indicated that competition-induced androstenedione and cortisol change were positively related. CONCLUSIONS Findings show that juvenile boys' steroid hormone responses during dyadic athletic competition differ in comparison to adult males, in whom cortisol and testosterone tend to rise. Lack of significant DHEA and androstenedione change during the table tennis competition differs from our previous work that showed DHEA and androstenedione were sensitive to more physically taxing forms of athletic competition (eg, soccer). These results are discussed in light of potential factors that may have contributed to these differences.

Background/Aims: There is a lack of evidence in the literature supporting vaginal application of a combination hormone-containing cream for local and systemic symptom relief. This pilot study examined the extent of absorption of a single cream containing estriol, estradiol, progesterone, DHEA, and testosterone. Methods: A combination cream was administered to 12 postmenopausal women in two differing doses over two independent time periods. Following 28 days (arm 1) and an additional 14 days (arm 2), measurement of hormones in saliva and blood and measurements of symptom relief, patient tolerability, and health-related quality of life (HRQoL) were obtained. Results: The dosage and time of evaluation for study arm 1 was not ideal for providing documented increases in hormone levels. HRQoL measurements supported measured improvement in this arm. The second arm did document absorption of the various hormones when given vaginally. Conclusion: This study is the first documenting systemic absorption of multiple hormones by both saliva and blood as well as improvement of HRQoL. This therapy was generally well-tolerated with only 2 patients experiencing minor irritation, not necessitating discontinuation. Additional studies in larger numbers of patients will provide better knowledge for clinicians wanting to provide similar therapy at the lowest effective dose.

Objective To compare LH, FSH, P, and E 2 levels obtained from blood spot vs. plasma (single-visit study) and to determine whether blood spots can document circulating hormone levels during ovulatory cycles (menstrual-cycle study). Design Cross-sectional study. Setting Academic center. Patient(s) Women 18–35 years of age with regular menstrual cycles and no recent use of hormonal contraception. Intervention(s) Women contributed both a blood spot sample from a finger-stick and a plasma sample through venipuncture on a random day within their menstrual cycle (n = 100, single study visit). Five additional women were followed for an entire menstrual cycle with biweekly venipuncture and daily self-collected blood spot sampling. Samples were analyzed for FSH, LH, P, and E 2 . Main Outcome Measure(s) Correlation between blood spot and plasma levels. Result(s) Significant positive correlations were found between the blood spot and plasma samples in the single-visit study ( r 2 : FSH, 0.91; LH, 0.93; P, 0.83; and E 2, 0.70). Two of the 5 menstrual-cycle study women had ovulatory cycles based on P levels (>3 ng/mL) and an LH surge. Daily blood spot sampling was better able to document hormonal changes than biweekly venipuncture. Conclusion(s) Blood spot monitoring of FSH, LH, P, and, to a lesser extent, E 2 , appears to be as valid as traditional plasma assays for clinical research and care.

Testicular germ cell tumors (TGCT) are the most commonly occurring cancers in adolescent and young adult males in the U.S. Steroid sex hormones play a central role in the development of the testis. As proposed by the testicular dysgenesis syndrome hypothesis, the origins of TGCT are likely to be in utero or early in life, and to be a manifestation of disturbed prenatal testis development. However, no studies have provided direct, empirical evidence to date. Using an innovative linkage between the California birth records and cancer registry data, we conducted a population-based case-control study of neonatal hormones levels and risk of TGCT diagnosed at 0-19 year of age. We obtained archived neonatal dried blood spot (DBS) specimens from 370 TGCT cases (276 adolescent and young adults [AYA] aged 15-19 yrs at diagnosis; 94 0-4 yrs at diagnosis), and 344 age- and race/ethnicity-matched controls, born between 1982 and 2009. Liquid chromatography with tandem mass spectrometry was used to measure a panel of 17 sex steroids, glucocorticoids, and mineralcorticoids; 12 were present at detectable levels in the newborn DBS samples, including estrone (E1), estradiol (E2), estriol (E3), testosterone (T), dehydroepiandrosterone (DHEA), and androstenedione (A4). Logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI), adjusting for matching factors and age (in hours) of child at blood spot collection. A4, a precursor for T and E1, was positively associated with TGCT (OR: 1.71, 95% CI: 1.09-2.69). Analyses stratified by age group showed that this association was limited to AYA, and was of stronger magnitude in this group (OR: 2.33, 95% CI: 1.37-3.97). A similar, though weaker, trend was observed for T (ORoverall: 1.37, 95% CI: 0.86-2.19; ORAYA: 1.73. 95% CI: 1.00-3.00). There was no significant association of the other measured hormones with risk. In the first case-control study of TGCT with direct measures of neonatal hormone levels, we found that higher levels of T and A4 were associated with increased risk of TCGT, particularly among males diagnosed at 15-19 years of age. These results oppose the dominant theory in TGCT etiology, that TGCT is related to androgen insufficiency in utero, and provides an important link in the etiologic pathway of this increasingly common cancer. Citation Format: Libby Morimoto, David Zava, Katherine McGlynn, Frank Stanczyk, Joseph Wiemels, Xiaomei Ma, Catherine Metayer. Neonatal hormone levels and risk of testicular germ cell tumors (TGCT) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2263. doi:10.1158/1538-7445.AM2017-2263

We investigated the estrogenic activity of various environmental pollutants (xenobiotics), in particular the xenoestrogen o,p-DDT, and compared their effects with those of endogenous estrogens, phytoestrogens, and mycoestrogens on estrogen receptor binding capacity, induction of estrogen end products, and activation of cell proliferation in estrogen-sensitive human breast cancer cells in monolayer culture. We also quantified the levels of phytoestrogens in extracts of some common foods, herbs, and spices and in human saliva following consumption of a high phytoestrogen food source (soy milk) to compare phytoestrogen abundance and bioavailability relative to the reported xenoestrogen burden in humans. Results show that natural endogenous estrogens, phytoestrogens, mycoestrogens, and xenoestrogens bind estrogen receptor (ER) in intact cells, but demonstrate marked differences in their ability to induce end products of estrogen action and to regulate cell proliferation. All of the different classes of estrogens stimulated cell proliferation at concentrations that half-saturated ER, but only some classes were able to induce estrogen-regulated end products. Genistein, a common phytoestrogen found in soy foods, differed from the xenoestrogen DDT in its effects on cell proliferation and ability to induce estrogen-regulated end products. Moreover, we found that many of the foods, herbs, and spices commonly consumed by humans contain significant amounts of phytoestrogens, and consumption of soy milk, a phytoestrogen-rich food, markedly increases the levels of phytoestrogens in saliva. In conclusion, our in vitro results predict that a diet high in phytoestrogens would significantly reduce the binding of weak xenoestrogens to ER in target tissues in vivo.

Background: Correlation between circulating sex steroid levels and breast cancer has been controversial, with measurement of free, or bioavailable hormone rarely available. Salivary hormone levels represent the bioavailable fraction. To further elucidate the role of endogenous hormones in breast cancer, we aimed to assess correlation between salivary sex steroid levels and breast cancer prevalence. Methods: Salivary hormone levels of testosterone (T), Estradiol (E2), Progesterone (P), Estriol (E3), Estrone (E1), DHEAS and Cortisol (C) were measured by Enzyme Immunoassay (EIA) in 357 women with histologically verified breast cancer and 184 age-matched control women. Results: Salivary T and DHEAS levels were significantly lower in breast cancer cases vs. controls (27.2+13.9 vs. 32.2+ 17.5 pg/ml, p < 0.001 for T and 5.3+4.3 vs. 6.4+4.5 ng/ml, p = 0.007 for DHEAS). E2 and E1 levels were elevated and E3 levels were lowered in cases vs. controls. Conclusions: Salivary T levels, representing the bioavailable hormone, are significantly lower in women with breast cancer compared to age-matched control women. These findings support the protective role of biovailable testosterone in counteracting the proliferative effects of estrogens on mammary tissue. Background The risk of developing breast cancer is related to events of reproductive life and lifestyle factors that modify levels of endogenous sex hormones. Soon after the discovery of sex hormones, it was suggested that breast cancer risk was related to endogenous estrogen levels. Recent large prospective cohort studies on postmenopausal women make it clear that high levels of estrogens increase mammary gland proliferation and are associated with increased breast cancer risk [1]. Interestingly, several observations suggest that androgens may counteract the proliferative effects of estrogen and progestogen in the mammary gland. In cell cultures and animal experiments, androgens have been shown to exert anti-proliferative effects [2]. It has also been demonstrated that a negative association exists between breast cell proliferation and levels of free testosterone in

In this study we report on the content and bioactivity of plant (phyto) estrogens and progestins in various foods, herbs, and spices, before and after human consumption. Over 150 herbs traditionally used by herbalists for treating a variety of health problems were extracted and tested for their relative capacity to compete with estradiol and progesterone binding to intracellular receptors for progesterone (PR) and estradiol (ER) in intact human breast cancer cells. The six highest ER-binding herbs that are commonly consumed were soy, licorice, red clover, thyme, tumeric, hops, and verbena. The six highest PR-binding herbs and spices commonly consumed were oregano, verbena, tumeric, thyme, red clover and damiana. Some of the herbs and spices found to contain high phytoestrogens and phytoprogestins were further tested for bioactivity based on their ability to regulate cell growth rate in ER (+) and ER (-) breast cancer cell lines and to induce or inhibit the synthesis of alkaline phosphatase, an end product of progesterone action, in PR (+) cells. In general, we found that ER-binding herbal extracts were agonists, much like estradiol, whereas PR-binding extracts, were neutral or antagonists. The bioavailability of phytoestrogens and phytoprogestins in vivo were studied by quantitating the ER-binding and PR-binding capacity of saliva following consumption of soy milk, exogenous progesterone, medroxyprogesterone acetate, or wild mexican yam products containing diosgenin. Soy milk caused a dramatic increase in saliva ER-binding components without a concomitant rise in estradiol. Consumption of PR-binding herbs increased the progestin activity of saliva, but there were marked differences in bioactivity. In summary, we have demonstrated that many of the commonly consumed foods, herbs, and spices contain phytoestrogens and phytoprogestins that act as agonists and antagonists in vivo.

Experimental and epidemiologic studies support the view that soyfoods prevent cancer as well as diseases and symptoms associated with estrogen deficiency. Recent research suggests that the isoflavonoid genistein, a phytoestrogen found in abundance in soyfoods, may be one of the principal molecular components responsible for these health benefits. In this study we investigated the effects of a broad physiologically relevant concentration range of genistein on estrogen receptor (ER) binding, induction of the estrogen-regulated antigen pS2, and cell proliferation rate in ER(+) and ER(-) human breast cancer cells grown in vitro. Dose response to genistein was compared with that of estradiol, tamoxifen, and several other structurally similar iso- and bioflavonoids (e.g., equol, kaempferol, and quercetin). Our results revealed that genistein has potent estrogen agonist and cell growth-inhibitory actions over a physiologically achievable concentration range (10 nM-20 microM). Other flavonoids over the same concentration range were good estrogen agonists and poor cell growth inhibitors (equol) or poor estrogen agonists and potent growth inhibitors (kaempferol and quercetin). The growth-inhibitory actions of flavonoids were distinctly different from those of triphenyl antiestrogens like tamoxifen. In summary, our results reveal that genistein is unique among the flavonoids tested, in that it has potent estrogen agonist and cell growth-inhibitory actions over a physiologically relevant concentration range.

The preparation of monoclonal antibodies (MAbs) against the human milk fat globule membrane with preferential binding to breast carcinoma cells is described. Using BALB/c mouse myeloma cells; inter-specific, intra-strain, and inter-strain hybridomas were isolated that identified three different components of the human milk fat globule of approximately 46,000, and 70,000 daltons and a mucin-like glycoprotein complex (NPGP) ranging from 400,000 to over a million daltons, respectively. Three MAbs (BrE1, BrE2, BrE3) identified the latter component which consists of at least three different size molecules for which the aforementioned MAb's have different binding specificities. MAbs, BrE2 and BrE3, bound to normal breast epithelial cells but to a lesser extent than to tumors and only at the apical surface facing the lumen, while they bound breast carcinomas strongly, and often in the cytoplasm as well as on the surface. Higher concentrations of BrE3 were required to stain normal breast compared to breast tumors. BrE1 also stained breast carcinomas both on the surface and cytoplasmically but did not stain normal breast tissue. The MAb, Mc13, as well as the previously reported MAb McR2, both against the 70,000 dalton component, did not significantly stain either normal or cancerous breast tissue in histological sections but did bind significantly to cultured breast epithelial cells and to the milk fat globule membrane. The MAbs, Mc8 and Mc3, reported previously to be against the 46,000 dalton component, stained histologically only malignant breast tissue but only weakly; however, they bound strongly to intact breast carcinoma cells and breast cell membrane preparations with a radioimmunobinding assay. These MAbs should be useful in characterizing the surface of breast epithelial cells, studying surface alterations in malignancy, and possibly in breast cancer diagnosis and therapy.

Breast cancer is often hormone responsive, since growth or regression of tumors can often be modulated by appropriate endocrine manipulations. Estrogen and progesterone are major hormones involved in regulation of breast cancer tumor growth. Considerable insight into the mechanism of action of these hormones on tumor growth stimulation has been provided by demonstration of specific receptors for each. The inference that each hormone acts independently through its receptor to control tumor growth is belied by current studies which show that certain hormones are capable of regulating the receptor sites, metabolism, or nuclear translocation of others. This may begin to explain the complex hormonal interactions and requirements of normal and neoplastic breast tissues. Considerable progress has thus been made in understanding the basis for success of various ablative therapies. The pharmacologie actions of estrogens and progestins in causing breast tumor regression is much less well understood. The role of hormone receptor sites has not been established in the mechanism of tumor regression caused by these pharmacological therapies. Nevertheless, when estrogen receptors are absent in a tumor, we can with accuracy pre.dict that endocrine therapies will fail, whereas when ER is present the likelihood of a successful response to pharmacological or ablative therapy is high. Receptor sites seem to be a common denominator and useful marker for hormone dependence or hormone responsiveness, irrespective of their actual role in the tumor regression process. Further investigations into the receptor functions should lead to new approaches in the endocrine management of patients with breast cancer.

Multiple intratumoral tissue samples from the primary mass of 30 consecutive invasive breast cancer patients were assayed for estrogen receptor (ER) and progesterone receptor (PR) by the dextran-coated charcoal method following frozen section histopathological examination. Steroid receptor status of each sample was classified as positive (R+) or negative (R−), based only upon quantitative guide lines form the ER/PR results. Four out of 32 (12.5%) of the invasive cancers had an intratumoral sample classified as R+ and one sample as R−. R+ invasive ductal carcinomas has a highly significant degree of tubule formation ( P = 0.005) when compared with R− invasive ductal cancers. While the quantitative ER content ( r = 0.18) and the degree of quantitative variation in ER content ( P = 0.04) did not correlate with the tumor cellularity of the individual samples, tumor cellularity ( P = 0.005) and ER content ( P = 0.005) were lower in the samples from the tumor border than from the central tumor samples. Variations in ER and PR content may be found on a regional basis within a breast tumor mass resulting from heterogeneity of tumor subpopulations and/or differences in tumor cellularity.

The anti-estrogen tamoxifen binds cytoplasmic estrogen receptor and translocates it into the nucleus, provoking early estrogen responses; only later is normal estrogen action antagonized. We examined these effects in uteri of mature rats given 5 μg estradiol or 50 μg tamoxifen after ovariectomy- adrenalectomy, using ligand exchange assays developed in this laboratory for both cytoplasmic and nuclear estrogen receptors. We show that both estrogen receptor translocation and uterine weight gain are slower with tamoxifen than with estrogen. Twenty-four to 48 h after estradiol or tamoxifen injections cytoplasmic estrogen receptor is replenished, contrary to the hypothesis that anti-estrogens antagonize subsequent estrogen action by failing to replenish cytoplasmic receptor. This new receptor appearing by 48 h after tamoxifen treatment is capable of being translocated by estradiol. Six daily doses of estradiol continue to increase uterine weight and soluble cytosol proteins. The same regimen of daily tamoxifen ...

Primary meningiomas have been grown in monolayer culture and tested for the presence of steroid hormone receptors and sensitivity to various steroids and steroid antagonists. None of the 10 solid tumors or the primary cultures derived from them contained estrogen receptors, either in the cytoplasm or in the nucleus. Progesterone receptors were present in 50-70% of the solid tumors and some of the primary cultures. Four of four and five of five primary cultures contained, respectively, androgen and glucocorticoid receptors. When one of the primary cultures was tested for growth sensitivity to estrogen, tamoxifen, progesterone, hydrocortisone, and dihydrotestosterone, the last two had noticeable stimulatory effects on growth by day 5. Interestingly, only androgen and glucocorticoid receptors were present in the primary tumor cells in culture, suggesting that these receptors mediated the effects of their respective hormones on growth.

Daily injections of estradiol or the antiestrogen tamoxifen initially stimulate uterine weight increase and progesterone receptor synthesis, though continued tamoxifen fails to maintain the increased weight. The stimulatory actions of both estradiol and tamoxifen are inhibited or reversed by a single injection of progesterone. It has been hypothesized that progesterone antagonizes estrogen action by reducing estrogen receptor levels, but in the present experiments neither cytoplasmic nor nuclear estrogen receptor was affected. We conclude that progesterone acts at a point beyond estrogen receptor availability or translocation to antagonize estrogen action.

Steroidal pyrimidines (steroid-nucleosides) have been suggested as potential agents for cancer chemotherapy. In order to evaluate the notion that the steroid moiety of such agents provides organ specificity, a series of steroid-nucleosides have been evaluated for their steroid binding properties. The 21-substituted progesterone derivatives showed affinity for androgen receptor whereas some of the 21-substituted corticoids exhibited affinity for glucocorticoid receptor. None of the 3- or 21-substituted steroids showed significant binding to estrogen or progesterone receptor.

Estradiol and progesterone receptors were studied in 44 patients with meningiomas and correlated to the clinicopathological features and amount of preoperative corticosteroid therapy. Thirty-four (77%) of the meningiomas contained high titers of specific high-affinity cytosol [3H]promegestone (R 5020) binding sites (mean 2,902 fmol/g tumor; range 0-9,598 fmol/g tumor) whereas only miniscule amounts of a nonspecific cytoplasmic [3H]estradiol binding component (mean 48 fmol/g tumor; range 0-201 fmol/g tumor) were detectable. No nuclear binding activity for [3H]estradiol was demonstrable. There was no convincing correlation between high PR activity and the age, sex, or menopausal status of the patients. The correlation study between the amount of preoperative corticosteroid therapy with the amount of [3H]promegestone binding revealed no dose relationship. Correlating [3H]promegestone content with the histologic type, we found 96% of meningothelial, 71% of transitional, and 40% of fibroplastic meningiomas to contain progesterone receptors. The necessity of in vitro studies is stressed to assess the biosynthesis and biological activity of the progesterone receptor in meningiomas, which is apparently not estrogen regulated, as is the case in other estrogen target tissues.

The reproducibility of oestrogen and progestin receptor assays performed by laboratories participating in an international breast cancer treatment trial has been assessed. Three tissue reference powders containing low, medium and high oestrogen receptor levels (22 ± 4, 88 ± 7, 227 ± 13 fmol/mg cytosol protein respectively) were prepared in Louisville, KY, U.S.A., assayed repeatedly and multiple samples of each shipped on solid CO2 to the coordination-distribution centre in Berne, Switzerland. Samples were dispatched from Berne to Cantons within Switzerland, to Yugoslavia, South Africa, Australia, New Zealand and also back to the United States for oestrogen and progestin receptor assays. Results were returned to Berne, Switzerland. There was a decrease in the levels of oestrogen and progestin receptors during the time of storage and transit. However, the ability to assign a powder to either the low, medium or high level of oestrogen receptor was not affected. Laboratories also determined progestin receptor. All laboratories clearly identified the powder containing the low level of progestin receptor, but there was poor quantitation with the other two assay standards. It is recommended that clinical hormone receptor laboratories, especially those participating in clinical trials, establish regular quality control procedures for both daily evaluation internally and periodic outside monitoring of interlaboratory variation.

Tumor estradiol and progesterone binding sites were studied in 34 patients with meningioma. Twenty of the meningiomas contained very low titers (mean, 45 fmol/g of tumor; range, 0-201 fmol/g of tumor) of a nonspecific cytoplasmic [3H]estradiol binding component, whereas 26 of the tumors contained high titers of specific high-affinity cytosol [3H]promegestone (R5020; progesterone) binding sites (mean, 1476 fmol/g of tumor; range, 0-8328 fmol/g of tumor). No nuclear binding activity for [3H]estradiol could be detected in 12 of the 34 meningiomas studied, irrespective of the progesterone binding activity. There was no correlation between high progesterone binding activity and the age or the sex of the patient, nor between tumor location and cellular mitotic index. However, progesterone binding activity was present more frequently in meningothelial (95%, 18/21 patients) than in transitional (55%, 5/9 patients) or fibroplastic (25%, 1/4 patients) tumor histologic types. These data suggest that the cellular biosynthesis of the progesterone binding component in meningiomas is not estrogen regulated as it is in other classic estrogen target tissues, such as the breast.

The synthetic radiolabeled androgen methyltrienolone-17βhiydroxy-17α-methyl-estra-4,9,ll-trien-3-one (R1881) is superior to the native ligand 5α-dihydrotestosterone (DHT) in the measurement of androgen receptor (AR), especially in tissues where radiolabeled DHT binds with high affinity to plasma proteins in addition to receptor or is rapidly metabolized to inactive derivatives. However, R1881 also binds progesterone receptor (PgR), as shown by its ability to complete for [3]-promestone-17α,21-dimethyl-19-nor-pregna-4,9-diene-3,20-dione (R5020) binding to 8S PgR on sucrose density gradients, resulting in an overestimation of AR if PgR is present. Therefore, we have devised a method by which PgR binding of R1881 can be blocked without interfering with AR binding. The glucocorticoid triamcinolone acetonide (TA) was found to bind PgR (competition for 8S [3]R5020 binding) but not AR (no competition for 8S [3H]DHT binding). In cytosols containing both receptors, a 500-fold excess of unlabeled TA was sufficient ...

Publisher Summary This chapter describes the methods for assessing the binding of steroid hormones in nuclei. When a steroid hormone enters a target cell, it binds to a receptor molecule that is then translocated into the nucleus. There it associates with nuclear sites and stimulates a sequence of events resulting in a series of biological responses. The binding of receptor-steroid complexes by nuclei can be studied by several methods. A radioactive steroid can be administered to intact animals or to cell or organ cultures, the nuclei isolated, and the content of radioactivity then measured directly. Nuclei can be isolated from cells and then incubated with radioactive receptor–steroid complexes prepared independently. Nuclei can be isolated and the receptor–steroid complexes measured by incubation with an excess of radioactive steroid at an elevated temperature permitting exchange with the unlabeled steroid. The difficulty of this assay is the technical problem of high nonspecific steroid binding in nuclei of tissues. To circumvent this problem, an alternative nuclear exchange assay is discussed in the chapter. Some of the methods developed and used to examine the binding of steroid receptors to isolated nuclei are also discussed.

Specific uptake of tritiated 17β-oestradiol and R5020, a synthetic progestin, in breast cancer cell lines (MCF7 and T47D) growing in monolayer culture in multiwell plates has been shown. Binding characteristics, calculated by Scatchard analysis, indicate the presence of steroid receptors of similar affinities and capacities to those already obtained with broken cell preparations. Lysis of the cells by treatment with a hypotonic buffer reveals the subcellular localization of the receptors so the method can be used to study receptor dynamics such as nuclear translocation and processing. Cell growth can be measured by DNA determination directly in the multiwell plates. Thus, the method provides a convenient way of studying the effects of steroid hormones (or any antihormone or chemotherapeutic agent) on growth and receptor content of breast cancer cells in monolayer culture.

The use of cytoplasmic estrogen receptor to predict endocrine responsiveness in breast cancer patients is now well established. It is our contention that this pertinent clinical application is only the first of many contributions from receptor studies to the clinic. Efforts are now underway to examine the distribution of receptors within tumor cells, and in particular the significance of receptors in tumor nuclei in the absence of hormone. Also, the demonstration of a direct stimulation of progesterone receptor synthesis by estradiol in cultured breast tumor cells now permits a careful dissection of the complete estrogen response system which should provide clues to the mechanism of antiestrogen action in causing tumor regression. Finally, it is now evident that pharmacologic effects of steroids in breast cancer may be mediated through different pathways than expected from studies of normal physiology.

A model system is presented to explain how tyrosinase, an enzyme unique to pigmented cells such as normal and malignant melanocytes can oxidize [3H]-estradiol to radiolabeled products which closely resemble the tight binding of [3H]-estradiol to estrogen receptor. In the model system studied, tyrosinase oxidized [2,4,6,7-3H]-estradiol to [3H]-water and [3H]-estradiol metabolites, the latter of which formed ring-substituted conjugates with nucleophiles like monothioglycerol and BSA. Radiolabeled estradiol without tritium in the C-2 position (i.e. [6,7-3H]-estradiol) failed to liberate [3H]-water when exposed to tyrosinase but, nevertheless, did form ring-substituted [3H]-estradiol adducts with nucleophiles. The [3H]-water and the ring-substituted radiolabeled products possessed several characteristics of a genuine estrogen receptor protein in that they were resistant to dextran-coated charcoal (DCC) adsorption and their enzymatic formation was inhibited with non-steroidal estrogens like diethylstilbestrol. Other natural (estradiol) and synthetic (hydroxytamoxifen) estrogens which contain the phenol grouping also inhibited the enzymatic oxidation of [3H]-estradiol. Although it was difficult to differentiate estrogen receptor from tyrosinase using the conventional DCC assay system, several differences in these two proteins permitted a distinction to be made between them. First, tyrosinase oxidation of [3H]-estradiol was markedly inhibited by sulfhydryl reducing agents (monothioglycerol) that stabilize [3H]-estradiol binding to estrogen receptor. Second, estrogen receptor adsorbed by hydroxylapatite whereas tyrosinase did not, thus permitting the separation of these two proteins prior to incubation with [3H]-estradiol. We conclude that the [3H]-estradiol binding components in melanoma previously reported to be estrogen receptor probably represent instead the radiolabeled products of the tyrosinase-catalyzed oxidation of [3H]-estradiol.

Androgens stimulate the growth of the human breast cancer cell line MCF-7. Very high doses are required, however, and therefore, we inquired whether they might exert this effect through some mechanism other than the androgen receptor. Evidence is presented here that it is actually the estrogen receptor, activated by weak but specific binding of androgens, which mediates this effect. Physiological 5α-dihydrotestosterone (DHT) at 10-8 M translocates androgen receptor to cell nuclei without affecting other steroid receptors or stimulating cell growth. High DHT (10-6 M) translocates estrogen as well as androgen receptor and stimulates cell growth. Receptors for progestins and glucocorticoids are not affected. Testosterone, 3β-androstanediol, and the antiandrogen, R2956, at 10-6 M also translocate the estrogen receptor, while progesterone, hydrocortisone, and the inactive isomer, 5β-DHT, have little or no effect. 5α-DHT at 10-6 M competes with estradiol for binding to cytosol estrogen receptor, while 10-8 M DH...

The role of cytoplasmic estrogen receptor (ER) assays in determining therapeutic strategies for advanced breast cancer is certainly well established. The use of ER assays in the primary breast tumor specimen to predict for early recurrence and ultimate survival is a new finding, however, and will probably be employed in future trials of adjuvant therapy. The prevalence and significance of nuclear-bound ER still requires additional clarification. Our previous suggestion that progesterone receptor measurements might be a useful marker for hormone dependence in advanced breast cancer is gaining support and may soon have a place in routine therapeutic decision-making. The emphasis on early adjuvant therapy has hastened the search for a safe endocrine therapy that would have good patient compliance and achieve remission rates comparable to previous agents and procedures. Antiestrogens show promise of meeting these requirements. We are now beginning an era in which primary and secondary systemic therapies for breast cancer can be based on sound biologic principles. The empirical approach is out-dated.