Research in Review

Winter 2003

Aeron LifeCycles Clinical Laboratory applies salivary analysis to the life long hormonal needs of women and men. As a tool for both interest and education, Aeron is compiling a quarterly review for clinicians, which will report short synopses of current findings in hormone science from peer reviewed publications. 

Diurnal Rhythm of Free Estradiol During the Menstrual Cycle. Salivary estradiol was measured by RIA in samples collected every 2h for 24hr from 15 normally cyclic healthy women during reproductive life (at different the three main phases of the cycle). They concluded that the diurnal rhythm of estradiol has a similar complex and temporal organization for different menstrual phases. The menstrual cycle mainly modulates the acrophase of the diurnal rhythm.
Eur J Endocrinol. 2003 Feb;148(2):227—32

Estrogen Fluctuations, Oral Contraceptives, and Borderline Personality. This paper combined the results from three studies that suggest fluctuation in estrogen level may influence the expression of borderline personality disorder (BPD) symptoms. They found that for women with high pre-existing levels of BPD, symptoms became significantly worse after starting an oral contraceptive. Research findings that link the serotonin system and estrogen are reviewed and theoretical and practical implications are discussed.
Psychoneuroendocrinology. 2003 Aug;28(6):751-66

Body Mass Index, Serum Sex Hormones, and Breast Cancer Risk in Postmenopausal Women. Obesity is associated with increased breast cancer risk among postmenopausal women. The study examined whether this association could be explained by the relationship of body mass index (BMI) with serum sex hormone concentrations. They concluded that the risk did increase with BMI and that this increase in relative risk was substantially reduced by adjustment for serum estrogen concentrations. These results are compatible with the hypothesis that the increase in breast cancer risk with increase BMI among post menopausal women is largely the result of the associated increase in estrogens, particularly bioavailable estradiol
J Natl Cancer Inst. 2003 Aug 20;95(16):1218-26

The Effects of a Low-Fat/High-Fiber Diet on Sex Hormone Levels and Menstrual Cycling in Premenopausal Women: a 12-Month Randomized Trial (The Diet and Hormone Study). A total of 213 healthy women, ages 20-40 years, were randomly assigned to follow their usual diet or to adopt an isocaloric diet with goals of 20% calories as fat, total fiber of 25g/day, and at least 8 fruit or vegetable servings per day. Total estradiol, SHBG, free estradiol, and progesterone were evaluated at baseline, after 4 cycles and after 12 cycles following the dietary intervention. Serum was collected 7-9 days after the LH surge. Serum estradiol decreased an average of 7.5% after 12 cycles in the intervention group vs. a decrease of 0.9% in the control group. They concluded that this 7.5% decrease in estradiol could have a biologic significance if it persisted over many years.
Cancer. 2003 Nov 1;98(9):1870-9

Analysis of the Progesterone Displacement of its Human Serum Albumin Binding Site by Beta-Estradiol using Biochromatographic Approaches: Effect of Two Salt Modifiers. Researchers studied the mechanisms of the binding of progesterone and 17-beta estradiol to human serum albumin (HSA) and progesterone’s displacement of its HSA binding cavity by 17-beta estradiol. They found a negative Gibbs free energy value which indicates that beta estradiol displaced progesterone of its HSA binding cavity, Moreover, the effect of two chloride modifiers [(Na(+), Mg(2+)] on these binding processes were analyzed. They discovered that the Mg (2+) cation enhanced strongly the bioavailable progesterone. Magnesium supplementation may benefit women who suffer from "estrogen dominance syndrome"
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Nov 5;796(2):267-81

Sex Hormone Levels and Risk of Cardiovascular Events in Postmenopausal Women. 200 women who were matched 1:1 by age, smoking, and postmenopausal hormone therapy (HT) to controls who remained free of CVD. Among HT nonusers, cases had significantly higher androgen profiles (Higher median FAI and lower SHBG levels) than controls. This was not independent of BMO and other cardiovascular risk factors. These researchers concluded that estradiol levels were not associated with risk of CVD in HT users or nonusers.
Circulation. 2003 Oct 7;108(14):1688-93

A Pilot Study on the Effects of Testosterone in Hypogonadal Aging Male Patients with Alzheimer’s Disease. The male aging process brings about declines in hormonal function including a gradual decline in bioavailable testosterone levels. This study from the University of Texas took 36 male patients with a new diagnosis of Alzheimer’s disease and measured their bioavailable testosterone levels. 10 were determined to be hypogonadal. 5 received 200mg of intramuscular T every two weeks, 5 received placebo. In the testosterone treated group, bioavailable testosterone increased from of mean of 48.7ng/dl to 142ng/dl. ADAScog (Alzheimer’s Disease Assessment Scale cognitive subscale), MMSE (Mini Mental Status Examination), CDT (Clock Drawing Test) all scores improved in the T treated group. The placebo-treated group deteriorated gradually.
Aging Male. 2003 Mar;6(1):13-7

Bioavailable Testosterone with Age and Erectile Dysfunction. Currently bioavailable testosterone (BT) is considered the most reliable marker for establishing the presence of hypogonadism. This study sought to clarify the relationship between BT and other hormones with respect to patient age and symptoms of partial androgen deficiency of the aging male (PADAM). Although serum total testosterone did not decrease with ages, SHBG increase significantly. BT and free testosterone decreased significantly. The International Index of Erectile Function-5 score for erectile function increased significantly with increases in BT.
J Urol. 2003 Dec;170(6 Pt 1):2345-7

Contribution of Bioavailable Testosterone Assay for the Diagnosis of Androgen Deficiency in Elderly Men. Testosterone levels in men decline progressively as of the thirties, at a rate which remains constant throughout life. Higher LH levels, decreased testosterone response to HCG and less Leydig cells all indicate that aging induces partial testicular failure. However, the gonadotropic function is also affected in aging. The LH response to GnRH is blunted compared to the situation in young men, therefore LH levels are not a valid index for androgen deficiency in elderly males. None of the androgen-dependent functions (libido, erection, and strength) are under exclusively androgen control, and there is no elderly male symptom, which is completely specific to androgen deficiency. Thus, in elderly men, when clinical symptoms might indicate androgen deficiency, biological confirmation is needed. Bioavailable testosterone assay by ammonium sulfate precipitation is the optimum method for diagnosing androgen deficiency.
Ann Endocrinol (Paris). 2003 Apr;64(2):117-25

Clomiphene Increases Free Testosterone Levels in Men with both Secondary Hypogonadism and Erectile Dysfunction: Who Does and Does Not Benefit. Hypogonadism produces by functional suppression of pituitary gonadotropins has been shown to correct with clomiphene citrate, but with modest effect on sexual function. This study looked at whether longer treatment would produce improved results. 178 men with secondary hypogonadism and ED received clomiphene citrate for 4 months. Sexual function improved in 75% with no change in 25%, while significant increases in LH and free testosterone occurred in all patients. Multivariable analysis showed that responses decreased significantly with aging. Men with anxiety related disorders responded better to normalization of testosterone. Assessment of androgen status should be accomplished in all men with ED.
Int J Impot Res. 2003 Jun;15(3):156-65

Testosterone and Gonadotropin Levels in Men with Dementia. The Oxford Project to Investigate Memory and Aging previously found lower levels of testosterone in men with AD compared with controls. In this study they assessed levels of pituitary gonadotropins that regulate sex steroid levels to determine whether primary of secondary hypogonadism was responsible for low levels of testosterone in cases. They found no difference in average LH, FSH or SHBG levels between AD cases and controls. T levels were significantly lower in men with AD and results were unchanged when controlled for age, SHBG and gonadotropin levels. They concluded that secondary hypogonadism occurs in men with AD. This is contrary to an earlier study (Bowen 1999) that found raised levels of gonadotropins in cases with AD – suggesting primary hypogonadism. Oxford’s cohort was younger and gonadotropin levels increase with age. They are enlarging their data set – to investigate whether secondary hypogonadism precedes cognitive dysfunction in men at risk for AD.
Neuroendocrinol Lett. 2003 Jun-Aug;24(3-4):203-8

Men in Committed, Romantic Relationships have Lower Testosterone. 122 male Harvard Business School students filled out a questionnaire and collected one saliva sample (from which testosterone was measured). Results revealed that men in committed, romantic relationships had 21% lower testosterone levels than men not involved in such relationships.
Horm Behav. 2003 Aug;44(2):119-22

Testosterone Replacement Therapy Restores Normal Ghrelin in Hypogonadal Men. This study investigated the interaction between sex steroids and ghrelin; we investigated circulating ghrelin levels in a group of hypogonadal men before and after therapeutic intervention aiming at normalization low testosterone (T) concentrations. Total and free T and plasma ghrelin levels were significantly lower in the hypogonadal men than in the control groups. Hypogonadal men also had a significantly higher insulin resistance state. Ghrelin levels were positively correlated with both total and free T concentrations. After a 6-month replacement T therapy, ghrelin levels of hypogonadal patients increased and did not differ significantly in comparison with both control groups. Ghrelin may exert a relevant role in the endocrine network connecting the control of the reproductive system with the regulation of energy balance.
J Clin Endocrinol Metab. 2003 Sep;88(9):4139-43



Midnight Salivary Cortisol versus Urinary Free and Midnight Serum Cortisol as Screening Tests for Cushing’s Syndrome. The determination of late night salivary cortisol levels has been reported to be a sensitive and convenient screening test for CS. The aim of this study was to compare the diagnostic performance of midnight salivary cortisol (MSC) measurement with the of midnight serum cortisol (MNC) and urinary free cortisol (UFC) in differentiating 41 patients with CS from 33 patients with pseudo-Cushings (PC), 199 with simple obesity and 27 normal weight volunteers. The overall diagnostic accuracy with saliva was 93% with a 95% confidence interval. They concluded that salivary cortisol measurement can be recommended as a first line test for CS in both low risk and high risk patients.
J Clin Endocrinol Metab. 2003 Sep;88(9):4153-7

An Investigation into the Relationship between Salivary Cortisol, Stress, Anxiety and Depression. This study examined the relationship between indices of self-reported emotional distress and absolute versus change in cortisol levels. 54 women attending a diagnostic breast clinic completed scales measuring stress, anxiety and depression and provided 5 saliva samples over the course of a single day for the measurement of cortisol. Analysis of the change in cortisol levels revealed a non-linear interaction effect between stress and anxiety and time of day.
Biol Psychol. 2003 Feb;62(2):89-96

Effect of Controlled-Release Melatonin on Sleep Quality, Mood, and Quality of Life in Subjects with Seasonal or Weather Associated Changes in Mood and Behavior. 58 healthy adults exhibiting subsyndromal seasonal affective disorder (s-SAD) and/or the negative or positive type of weather-associated syndrome (WAS) were randomized to either 2mg of sustained release melatonin or placebo tablets 1-2h before bedtime for 3 weeks. Early morning salivary melatonin concentrations were measured at baseline and treatment cessation in all subjects. Melatonin administration significantly improved the quality of sleep and vitality in subjects with s-SAD, but attenuated the improvement of atypical symptoms and physical parameters of quality of life compared to placebo in subjects with WAS, positive type.
Eur Neuropsychopharmacol. 2003 May;13(3):137-45

Melatonin Phase-Shifts Human Circadian Rhythms with no Evidence of Changes in Duration of Endogenous Melatonin Secretion or the 24-hour Production of Reproductive Hormones. This study investigated the effects of an artificially prolonged melatonin profile on endogenous melatonin and cortisol rhythms, wrist actigraphy, and reproductive hormones in humans. This was a double blind crossover study using 8 healthy men. Surge/sustained release melatonin (1.5mg) or placebo was administered for 8 d at the beginning of a 16h sleep opportunity in dim light. Compared with placebo, melatonin administration advanced the timing of endogenous melatonin and cortisol rhythms. Melatonin treatment did not affect the endogenous melatonin profile duration, pituitary/gonadal hormone levels, or sleepiness and mood levels on the subsequent day.
J Clin Endocrinol Metab. 2003 Sep;88(9):4303-9

Alterations in the Circadian Rhythm of Salivary Melatonin Beginning during Middle Age. 52 healthy volunteers participated in the study consisting of young, middle aged, old and oldest groups. Time point salivary melatonin samples were taken over a 24-hour period. A clear circadian rhythm of salivary melatonin was present in all age groups. The decline in nocturnal peak levels in salivary melatonin was found in old and the oldest subjects. Both old and oldest subjects showed an increased daytime melatonin level. They found a step-wise decrease in the circadian rhythms of saliva melatonin occurred early in life – around 40 years of age. The middle-aged subjects had only 60% of the amplitude of the young subjects.
J Pineal Res. 2003 Jan;34(1):11-6

Longitudinal Association between Sex Hormone Levels, Bone Loss and Bone Turnover in Elderly Men. 200 elderly men (aged 55-85 yr) were followed for 4 years to evaluate the relationships between hormone levels, bone turnover markers, bone mineral density, and rates of bone loss. In the total population, T, calculated bioavailable T, calculated bioavailable estradiol, free androgen index, and free estradiol index, but not estradiol, decreased significantly with age, whereas SHBG increased significantly. Subjects with FEI, c-bioE and E levels below the median showed higher rates of bone loss. The ratio between estradiol and testosterone (and indirect measure of aromatase activity) increased significantly with age and was higher in normal than in osteoporotic subjects.
J Clin Endocrinol Metab. 2003 Nov;88(11):5327-33

Effects of a Triphasic Combination Oral Contraceptive Containing Norgestimate/Ethinly Estradiol on Biochemical Markers of Bone Metabolism in Young Women with Osteopenia Secondary to Hypothalamic Amenorrhea. This multicenter, double-blind, placebo-controlled, randomized study of 45 patients evaluated the short term effects of an oral contraceptive on biochemical markers of bone resorption in young women (mean age 26.5) with hypothalamic amenorrhea and osteopenia. The study used deoxypyridinoline, N-telopeptide, and bone specific alkaline phosphatase as biomarkers of bone metabolism. The results suggest that Ortho Tri Cyclen (250mcg norgestimate and 35mcg of ethinyl estradiol) decreases bone turnover in osteopenic premenopausal women with hypothalamic amenorrhea. Further studies are needed to determine whether estrogen will increase bone density in this population.
J Clin Endocrinol Metab. 2003 Aug;88(8):3651-6