Hormonal Profiles of Menstrual Bleeding Patterns During the Luteal-Follicular Transition.

CONTEXT: Menstrual cycle function is determined by a complex endocrine axis that controls the ovaries and endometrium. While the late luteal phase is characterized by declining progesterone and estrogen, how these hormonal profiles relate to menstrual bleeding patterns is not well understood. OBJECTIVE: Characterize associations between luteal phase hormonal profiles and subsequent menstrual bleeding patterns, specifically spotting before bleeding. DESIGN, SETTING, AND PARTICIPANTS: We examined creatinine-adjusted urinary estrone 3-glucuronide (E13G) and pregnanediol 3-glucuronide (Pd3G) levels in relation to spotting in 116 premenopausal women (ages 20-47) who kept daily menstrual diaries and collected first morning urine samples for ≥ 2 consecutive cycles or 1 luteal-follicular transition (n = 283 transitions). We used linear mixed models to estimate associations between luteal phase hormone levels and spotting before bleeding. MAIN OUTCOME MEASURE(S) AND RESULTS: Transitions with ≥ 1 days of spotting before menstrual bleeding (n = 118) had greater luteal phase Pd3G levels vs nonspotting transitions (n = 165). Differences in Pd3G between spotting and nonspotting transitions were largest at menses onset (34.8%, 95% confidence interval, 18.9%, 52.7%). Pd3G levels for spotting transitions dropped to similar levels as nonspotting transitions an average of 1 day later, which aligned with the first day of bleeding for transitions with contiguous spotting. Spotting transitions were preceded by slower rates of Pd3G decline than nonspotting transitions, whereas E13G declines were similar. CONCLUSIONS: Self-reported bleeding patterns may provide insight into luteal phase Pd3G levels. First bleed appears to be the best choice for defining the end of the luteal phase and achieving hormonal consistency across transitions.

Multiplex Surface-Enhanced Raman Scattering Identification and Quantification of Urine Metabolites in Patient Samples within 30 min.

Successful translation of laboratory-based surface-enhanced Raman scattering (SERS) platforms to clinical applications requires multiplex and ultratrace detection of small biomarker molecules from a complex biofluid. However, these biomarker molecules generally exhibit low Raman scattering cross sections and do not possess specific affinity to plasmonic nanoparticle surfaces, significantly increasing the challenge of detecting them at low concentrations. Herein, we demonstrate a "confine-and-capture" approach for multiplex detection of two families of urine metabolites correlated with miscarriage risks, 5ß-pregnane-3α,20α-diol-3α-glucuronide and tetrahydrocortisone. To enhance SERS signals by 1012-fold, we use specific nanoscale surface chemistry for targeted metabolite capture from a complex urine matrix prior to confining them on a superhydrophobic SERS platform. We then apply chemometrics, including principal component analysis and partial least-squares regression, to convert molecular fingerprint information into quantifiable readouts. The whole screening procedure requires only 30 min, including urine pretreatment, sample drying on the SERS platform, SERS measurements, and chemometric analyses. These readouts correlate well with the pregnancy outcomes in a case-control study of 40 patients presenting threatened miscarriage symptoms.

A Quadriparametric Model to Describe the Diversity of Waves Applied to Hormonal Data.

BACKGROUND: Even in normally cycling women, hormone level shapes may widely vary between cycles and between women. Over decades, finding ways to characterize and compare cycle hormone waves was difficult and most solutions, in particular polynomials or splines, do not correspond to physiologically meaningful parameters. OBJECTIVE: We present an original concept to characterize most hormone waves with only two parameters. METHODS: The modelling attempt considered pregnanediol-3-alpha-glucuronide (PDG) and luteinising hormone (LH) levels in 266 cycles (with ultrasound-identified ovulation day) in 99 normally fertile women aged 18 to 45. The study searched for a convenient wave description process and carried out an extended search for the best fitting density distribution. RESULTS: The highly flexible beta-binomial distribution offered the best fit of most hormone waves and required only two readily available and understandable wave parameters: location and scale. In bell-shaped waves (e.g., PDG curves), early peaks may be fitted with a low location parameter and a low scale parameter; plateau shapes are obtained with higher scale parameters. I-shaped, J-shaped, and U-shaped waves (sometimes the shapes of LH curves) may be fitted with high scale parameter and, respectively, low, high, and medium location parameter. These location and scale parameters will be later correlated with feminine physiological events. CONCLUSION: Our results demonstrate that, with unimodal waves, complex methods (e.g., functional mixed effects models using smoothing splines, second-order growth mixture models, or functional principal-component- based methods) may be avoided. The use, application, and, especially, result interpretation of four-parameter analyses might be advantageous within the context of feminine physiological events.

Menstrual Cycle Hormone Changes in Women Traversing Menopause: Study of Women's Health Across the Nation.

Context: Menstrual cycle hormone patterns in women approaching menopause are inadequately studied. Objective: To describe day-to-day menstrual cycle hormones in women as they approach menopause from the Study of Women's Health Across the Nation Daily Hormone Study (DHS). Design: DHS enrollees collected daily urine for one entire menstrual cycle or up to 50 days, whichever came first, annually, up to the final menstrual period (FMP) or for up to 10 years. Setting: Seven sites across the United States. Participants: A total of 511 premenopausal or early perimenopausal women at enrollment, within 10 years before menopause. Intervention: Time-to-FMP measurement. Main Outcome Measures: Evidence of luteal activity (ELA), determined using objective algorithms. Menstrual cycle/segment length; whole cycle, and segment integrated urinary luteinizing hormone, follicle-stimulating hormone, estrone conjugates, and pregnanediol glucuronide (Pdg) for each year, organized around the FMP. Results: Mean menstrual cycle length was remarkably preserved at 26 to 27 days in ELA cycles; non-ELA cycles had greater variability. The percentage of cycles that were ELA remained high until 5 years before the FMP (87.9%); only 22.8% of cycles within 1 year of the FMP were ELA. Whole cycle hormones remained relatively stable up to 3 years before the FMP, when gonadotropins began to increase. Pdg excretion declined slowly with progress to the FMP, but Pdg patterns of ELA cycles remained distinguishable from non-ELA. Conclusions: Menstrual cycle hormone patterns in perimenopausal women resemble those of midreproductive-aged women until 5 years before menopause, and presumably ovulatory cycles retain a potentially fertile pattern up to the end of reproductive life.

Puberty, ovarian cycle, pregnancy, and postpartum ovulation in captive Sichuan golden monkeys (Rhinopithecus roxellana) based on changes in urinary and fecal gonadal steroid metabolites.

The purpose of this study was to evaluate the reproductive status and clarify the reproductive physiology of captive Sichuan golden monkeys. The concentrations of urinary estradiol-3-glucuronide (E2G) and pregnanediol-glucuronide (PdG) or fecal estradiol-17ß (E2) and PdG in two females, and fecal testosterone concentrations in a male, were measured continuously using enzyme immunoassays. On the basis of these hormone profiles, the follicular phase, luteal phase, and ovarian cycle were calculated to be 14.7 ± 4.8, 10.4 ± 2.8, and 25.1 ± 3.3 days, respectively. The first ovulation (puberty) in a female monkey was observed at 5.1 years old, and the first pregnancy was diagnosed at 6.4 years old. For the first 2 months of pregnancy (204 days), fecal E2 and PdG maintained constant high values and then increased until parturition. These profiles were similar to urinary E2G and PdG changes. During the last trimester of a twin pregnancy, fecal PdG was up to approximately three times higher compared with a single pregnancy. Therefore, fecal PdG levels in late pregnancy may be effective for the detection of a twin pregnancy. The first postpartum ovulation occurred 66 (fetal death and artificial rearing), 143 (fetal death), and 189 (natural suckling) days after parturition. The anovulation period of the natural suckling case was longer than the others. Conception and postpartum ovulation were detected between September and January. Fecal testosterone levels of the male were correlated with the fecal E2 level of the nonpregnancy period in exhibited together female. Our results reported that urinary (E2G and PdG) and fecal (E2 and PdG) hormone measurement is effective for monitoring the reproductive status, thereby expanding knowledge of the reproductive endocrinology of this endangered species.

Validation of a field-friendly extraction and storage method to monitor fecal steroid metabolites in wild orangutans.

Measuring hormone metabolites from feces is the most often used method to assess hormonal status in wildlife. Although immediate freezing of fecal samples collected in the field is the best method to minimize the risk of degradation of hormones over time, this is often not possible in remote field sites. Therefore, alternative storage and preservation methods for fecal samples are required in these conditions. We conducted an experiment to investigate if fecal glucocorticoid (FGCM) and progesterone metabolite (pregnanediol-3-glucuronide; PdG) levels measured from samples that were extracted with a simple, field-friendly methodology correlate with those generated from frozen samples. We also evaluated whether storing fecal samples in alcohol is a suitable alternative to preserve FGCM and PdG concentrations long-term (i.e. over a 9-month period) at locations where fecal extraction is not feasible. Finally, we tested if the hormone concentrations in unpreserved fecal samples of orangutans change over 14 h when stored at ambient conditions, representing the maximum duration between sample collection and return to the camp. FGCM and PdG levels measured from samples that were extracted with the field-friendly method showed strong correlations with those generated from frozen samples, and mean levels did not differ significantly between these methods. FGCM concentrations showed no significant change compared to control samples when fecal samples were stored for up to 6 months in alcohol at ambient temperature and PdG concentrations even remained stable for up to 9 months of storage. FGCM concentrations of fecal samples kept at ambient temperature for up to 14 h post-defecation did not significantly differ compared to control samples frozen immediately after collection. These results provide the basis for the successful monitoring of the physiological status of orangutans living in remote natural settings, like those included in the Indonesian reintroduction programs.

Endocrine assessment of ovarian cycle activity in wild female mountain gorillas (Gorilla beringei beringei).

Variability of fertility (i.e. number of births per female per year) has been reported in females of many primate species but only a few studies have explored the associated physiological and behavioral patterns. To investigate the proximate mechanisms of variability in fertility of wild female mountain gorillas (Gorilla beringei beringei), we quantified the occurrence of ovulation, matings, and successful pregnancies among females. We examined the profiles of immunoreactive pregnanediol-3-glucuronide (iPdG) for sixteen females (seven nulliparous and nine parous females, including one geriatric female; average sampling period for fecal sample collection and behavioral observations per female=175 days; SD=94 days, range=66-358 days) monitored by the staff of the Dian Fossey Gorilla Fund's Karisoke Research Center in Parc National des Volcans, Rwanda. We quantified ovarian cycles from iPdG profiles using an algorithm that we developed by adjusting the method of Kassam et al. (1996) to the characteristics of ovarian cycle profiles based on fecal hormone measurements. The mean length of ovarian cycles was 29±4 days (median: 28 days, N=13 cycles), similar to ovarian cycle lengths of other great apes and humans. As expected, we found that female mountain gorillas exhibit longer follicular phases (mean±SD: 21±3 days, N=13 cycles) than luteal phases (mean±SD: 8±3 days, N=13 cycles). We also found that the frequency of ovarian cycles was greater in parous females (i.e. 20 ovarian cycles across 44 periods of 28 days; 45.5%) than in nulliparous females (i.e. two ovarian cycles across 34 periods of 28 days; 6%). However, the frequency of days on which matings were observed did not differ significantly between parous and nulliparous females, nor between pregnant and non-pregnant females. Five pregnancies were detected with iPdG levels, but only three resulted in live births, indicating miscarriages of the other two. In sum, this study provides information on the underlying endocrine patterns of variation in fertility depending on parity, mating behavior, and pregnancy success in a critically endangered great ape.

Endocrine pregnancy monitoring in the two-toed sloth (Choloepus didactylus): "Pregnant or not pregnant".

Progesterone (P4), pregnanediol glucuronide (PdG), estradiol-17ß (E2), and estrone sulfate (E1S) were measured in the feces of four female two-toed sloths (Choloepus didactylus) for early pregnancy diagnosis. For individual feces assignment, the examined female sloths were fed with a turquoise food colorant every second day. Fecal samples were collected one to four times per week, depending on the defecation rate throughout the pregnancies and the postpartum periods. The complete course of pregnancy was subdivided into three 16-week intervals (trimester of pregnancy, TP1-3) and a 5-week post-partum period after birth. Progesterone and PdG concentrations started to increase above luteal phase levels 3 weeks after conception (P = 0.028 and 0.005, respectively). At the beginning of TP1, P4 concentrations averaged 345.0 ± 283.0 ng/g and increased approximately 100- to 300-fold to a peak of 7588.0 ± 6717.0 ng/g over the TP3. Progesterone concentrations were considerably lower than PdG concentrations that started with 3206.0 ± 1500.0 ng/g at TP1 and increased up to 12.8556.0 ± 53.744.0 ng/g until birth. In contrast, mean concentrations of E2 (8.2 ± 2.4-11.7 ± 4.2 ng/g) and E1S (12.2 ± 6.7-22.9 ± 13.0 ng/g) elevated insignificantly and were not suitable for pregnancy detection. All hormones analyzed decreased rapidly within the first weeks after birth. Progesterone and PdG, as well as E2 and E1S, highly significantly correlated (r = 0.602, P < 0.001 and r = 0.497, P < 0.001, respectively) at TP1. During the TP2, only P4 and PdG significantly correlated (TP2: r = 0.661, P < 0.001 and postpartum period: r = 0.616, P = 0.009). In summary, only P4 metabolite concentrations were suitable to determine the status of reproduction in the two-toed sloth. Thereby, PdG was ideally suited to diagnose early pregnancy because it was more sensitive and detected pregnancy 2 weeks earlier than P4.

Cognitive dietary restraint: impact on bone, menstrual and metabolic status in young women.

We compared bone mineral density (BMD) and content (BMC), menstrual and metabolic status between physically active women with 1) high cognitive dietary restraint (High-CDR) (score > or = 9, n=38) and Normal-CDR (score<9, n=46) and 2) across quartiles of CDR scores. Eighty-four physically active (500+/-35 min wk(-1)) premenopausal women participated and were categorized according to their CDR score. Primary outcomes included, BMD, BMC, menstrual status, estrone-3-glucuronide (E1G) and pregnanediol-3-glucuronide (PdG) area under the curve (AUC). Secondary outcomes included resting energy expenditure (REE), total triiodothyronine, and ghrelin. Measures of body mass (59.2+/-1.1 vs. 58.5+/-1.0 kg) and percent body fat (24.7+/-1.2 vs. 23.7+/-0.7%) were similar between women with Normal-CDR and High-CDR, however the High-CDR group had lower total body (1.140+/-0.011 vs. 1.179+/-0.010 g cm(-2); p=0.015) and lumbar spine (1.114+/-0.019 vs. 1.223+/-0.022 g cm(-2); p=0.001) BMD. The prevalence of oligo-amenorrhea was higher in the High-CDR group and became increasingly greater across the CDR quartiles. There were no differences in metabolic characteristics between the High-CDR and Normal-CDR groups, however REE and the ratio of measured to predicted REE were lower in the fourth quartile (CDR scores > or = 13) compared to the second and third quartiles. Our results provide evidence that high CDR scores are associated with reduced lumbar spine and total body BMD in physically active premenopausal women. A greater frequency of menstrual disturbances in women with higher CDR scores likely played a role in the reduced total body and lumbar spine BMD.

Progesterone metabolites rapidly stimulate calcium influx in human platelets by a src-dependent pathway.

The effects of several steroids and their metabolites were examined for their ability to rapidly alter intracellular free calcium ([Ca(2+)](i)) in the anucleate human platelet. Earlier studies suggested that steroids had direct and rapid non-genomic effects to alter platelet physiology. The rationale for performing this study was to investigate the signal transduction events being activated by steroids. Super-physiologic concentrations (1.0-10.0microM) of beta-estradiol and several estradiol metabolites and analogs potentiated (approximately twofold) the action of thrombin to elevate [Ca(2+)](i) in platelets, whereas 10.0microM progesterone inhibited the action of thrombin by 10-15%. Progesterone and beta-estradiol by themselves did not affect [Ca(2+)](i). Progesterone metabolites can achieve high blood concentrations. Some progesterone metabolites, particularly those in the beta-conformation, were potent stimulators of Ca(2+) influx and intracellular Ca(2+) mobilization in platelets. They activated phospholipase C because their ability to increase [Ca(2+)](i) was inhibited by the phospholipase C inhibitor U-73122. The ability of pregnanediol and collagen to increase [Ca(2+)](i) was inhibited by the src tyrosine kinase inhibitor PP1, whereas the actions of thrombin and thapsigargin to increase [Ca(2+)](i) were not affected by PP1. The effects of progesterone metabolites to increase [Ca(2+)](i) were observed with concentrations as low as 0.1microM. Pregnanolone synergized with thrombin to increase [Ca(2+)](i). It is hypothesized that human platelets possess receptors for progesterone metabolites. These receptors when stimulated will activate platelets by causing a rapid increase in [Ca(2+)](i). Pregnanolone, isopregnanediol and pregnanediol were the most effective stimulators of this newly identified src-dependent signal transduction system in platelets. Progesterone metabolites may regulate platelet aggregation and hence thrombosis in vivo.

Luteal phase estrogen is decreased in regularly menstruating older women compared with a reference population of younger women.

OBJECTIVE: To compare daily reproductive hormone secretion in regularly menstruating older versus younger women. DESIGN: This was a prospective cohort study. RESULTS: Daily morning urine samples were obtained from 106 women, 28 of whom were aged between 20 and 34 years (mean: 27.8+/-3.7 y) and 78 of whom were aged between 35 and 50 years (mean: 40.3+/-3.7 y). Lower luteal estrone-3-glucuronide levels were seen in the older versus the younger group (82.7 vs 93.5 ng/ml, P=0.035). The pregnanediol-3-glucuronide levels in the older group were lower than those in the younger group throughout the entire cycle. The median length of the follicular phase was shorter in the older versus younger women (13 vs 14.5 d, P=0.005). There was no significant difference in the median luteal phase lengths between groups. CONCLUSIONS: We report the new finding that regularly menstruating older women not only have lower pregnanediol-3-glucuronide levels but also have a significant reduction in luteal phase estrone-3-glucuronide compared with a contemporaneous cohort of younger women. This combined deficit may play a key role during the luteal-follicular transition, potentially affecting follicle recruitment and decreasing fecundity in the subsequent cycle.

Gonadotropin stimulation of steroid synthesis and metabolism in the Rana pipiens ovarian follicle: sequential changes in endogenous steroids during ovulation, fertilization and cleavage stages.

Steroid synthesis and metabolism have been followed in Rana pipiens ovarian follicles, denuded oocytes and eggs during ovulation, fertilization and cleavage stages (blastula formation). Under physiological conditions, gonadotropin stimulation of the fully grown follicle leads to progesterone synthesis from [(3)H]acetate as well as formation of much smaller amounts of 17alpha-hydroxyprogesterone, androstenedione, pregnanedione and pregnanediol. Progesterone levels increase during completion of the first meiotic division, but by ovulation progesterone disappears from the egg. Plasma membrane-bound progesterone is taken up into the oocyte cortical granules and is largely metabolized to 5alpha-pregnane-3alphaol,20-one and 5beta-pregnane-3alpha,17alpha,20beta-triol coincident with internalization of 60% of the oocyte surface (and >90% of bound progesterone) by the end of the hormone-dependent period. The principal steroid in the ovulated egg is 5beta-pregnane-3alpha,17alpha,20beta-triol. There is a rapid efflux of 5beta-pregnane-3alpha,17alpha,20beta-triol into the medium immediately following fertilization and residual steroid levels remain low in the developing blastula. Dissociated blastulae cells prepared from stage 9 1/2 embryos concentrate both pregnenolone and progesterone from the medium with minimal metabolism. The results indicate that the ovarian follicle has the ability to synthesize and metabolize progesterone but that this ability disappears in the ovulated egg. The progesterone metabolites formed during meiosis are largely released at fertilization.

Urinary sex steroid excretion levels during a soy intervention among young girls: a pilot study.

Soy intake early in life may protect against breast cancer later in life, possibly by altering sex hormone metabolism. We evaluated the feasibility of assessing urinary sex steroid excretion among 20 young girls aged 8-14 yr in an 8-wk trial. The girls consumed one daily soy serving, collected weekly overnight urine samples, and reported Tanner stages for breast and pubic hair development. Sex steroid excretion was measured in duplicate by gas chromatography-mass spectrometry and adjusted for urinary creatinine. The respective coefficients of variation for estrone, estradiol, estriol, testosterone, pregnanediol were 11.4%, 10.4%, 8.4%, 12.8%, and 4.6%. The statistical analysis included t-tests, Spearman's correlations, and analysis of variance. Seventeen girls completed the study and showed good compliance with the intervention strategy. We observed nonsignificant increases in total androgens (0.11 microg/mg creatinine) and total estrogens (0.001 microg/mg creatinine) and a nonsignificant decrease in pregnanediol (-0.03 microg/mg creatinine) during the study period. Higher Tanner stages for pubic hair development were associated with ninefold higher estrogen, fourfold higher androgen, and twofold higher pregnanediol excretions (P=0.01, P<0.001, and P=0.047, respectively). Similar differences were observed after stratification by breast development and menarcheal status. The association of sex steroid levels with pubertal development supports the validity of the sex steroid measurements.

Ovarian cycle phase and same-sex mating behavior in Japanese macaque females.

The relationship of the ovarian cycle phase to same-sex mounting activity in adult female Japanese macaques (Macaca fuscata) was studied during the 1997/1998 mating season. Fecal samples were collected from eight female subjects two to three times per week and analyzed by enzyme immunoassay for fecal hormone levels. Hormone profiles of estrone (E1) and pregnanediol (PdG) were used to separate ovarian cycles into three phases: follicular, periovulatory, and luteal. Patterns of same-sex and heterosexual mounting behavior in the females were analyzed for phase variation during conceptive cycles. Same-sex mounting among female Japanese macaques occurred most frequently during the follicular and periovulatory phases of the cycle, and not at all during the luteal phase, paralleling the pattern found in heterosexual mounting behavior. These findings suggest a link between hormonal fluctuations and patterns of sexual mounting, regardless of whether the partner is of the same or opposite sex.

5alpha-androstane-3alpha,17beta-diol is formed in tammar wallaby pouch young testes by a pathway involving 5alpha-pregnane-3alpha,17alpha-diol-20-one as a key intermediate.

The synthetic pathway by which 5alpha-androstane-3alpha,17beta-diol (5alpha-adiol) is formed in the testes of tammar wallaby pouch young was investigated by incubating testes from d 20-40 males with various radioactive precursors and analyzing the metabolites by thin-layer chromatography and HPLC. [(3)H]Progesterone was converted to 17-hydroxyprogesterone, which was converted to 5alpha-adiol by two pathways: One involves the formation of testosterone and dihydrotestosterone as intermediates, and the other involves formation of 5alpha-pregnane-3alpha,17alpha-diol-20-one (5alpha-pdiol) and androsterone as intermediates. Formation of 5alpha-adiol from both [(3)H]testosterone and [(3)H]progesterone was blocked by the 5alpha-reductase inhibitor 4MA. The addition of nonradioactive 5alpha-pdiol blocked the conversion of [(3)H]progesterone to 5alpha-adiol, and [(3)H]5alpha-pdiol was efficiently converted to androsterone and 5alpha-adiol. We conclude that expression of steroid 5alpha-reductase in the developing wallaby testes allows formation of 5alpha-reduced androgens by a pathway that does not involve testosterone as an intermediate.

Caution on the use of saliva measurements to monitor absorption of progesterone from transdermal creams in postmenopausal women.

OBJECTIVES: To determine the levels of progesterone in plasma, red cells and saliva as well as pregnanediol-3-glucuronide excretion in postmenopausal women using transdermal progesterone creams. METHODS: A double-blind placebo controlled study was carried out using 24 postmenopausal women. Creams (placebo, 20 or 40 mg progesterone/g) were applied twice daily for 3 weeks followed by 1 week without before a further 3-week treatment. Morning samples were collected at 0, 1, 3, 4, 7 and 8 weeks for analysis. RESULTS: There were small increases in plasma progesterone levels and pregnanediol-3-glucuronide excretion compared to the placebo group and red cell progesterone levels never exceeded plasma levels during progesterone cream use. Saliva progesterone levels were very high and variable in the progesterone cream groups compared to the placebo group and presented a paradox to the usual relationship observed between plasma and saliva progesterone in premenopausal women. CONCLUSION: The absorption of progesterone from transdermal creams is low and we caution against the use of saliva measurements to monitor progesterone absorption. The low systemic absorption of progesterone may not be due to peripheral conversion by 5 alpha-reductase(s). We also conclude that the low level of progesterone associated with red cells suggests they are not important in the delivery of progesterone to target tissues.

[Effects of exposure to low-level benzene and its analogues on reproductive hormone secretion in female workers].

OBJECTIVE: To study the effects of exposure to low level of mixed benzene on reproductive hormone secretion during menstrual cycle in female workers. METHODS: Concentrations of benzene, toluene and xylene in the expiratory air of 50 exposed female workers from their breathing zone were determined with gas chromatography. Their menstrual cycles of 50 exposed workers and 35 internal control and 35 external control workers were studied prospectively. Urine pregnandiol-3-glucuronide (PdG), follicle stimulating hormone (FSH) and estrone conjugate (E1C) were measured by enzyme immunoassay. RESULTS: Mixed benzene existed mainly in the form of low-level benzene in the air of workplaces, with a detection rate of 29.10% at an average level of 8.88 (0.90-876.47) mg/m3, and 21% of measurements exceeded the national maximum allowable concentration with the highest one as 20.91 folds high as that of the national hygienic standard. Length of luteal phase in the exposed group (13.7 +/- 1.5) days was significantly shorter than that in the internal and external control groups (14.5 +/- 1.2) days and (15.2 +/- 1.1) days (P < 0.05 or P < 0.01). Urine level of E1C before ovulation, that of FSH at early follicular phase and that of PdG in luteal phase after ovulation in the exposed group were significantly lower than those in the internal control group (P < 0.05 or P < 0.01). CONCLUSION: Exposure to low level of mixed benzene in workspaces could interrupt the function of hypothalamic-pituitary-ovarian axis and affect their normal levels of FSH, PdG and E1C.

Longitudinal gonadal steroid excretion in free-living male and female meerkats (Suricata suricatta).

Slender-tailed meerkats (Suricata suricatta) are small, diurnal, cooperatively breeding mongooses of the family Herpestidae. A prerequisite to fully understanding the mating system of meerkats is the development of a normative reproductive-endocrine database. This study examined longitudinal gonadal steroid excretion in all adult and juvenile individuals of both sexes within a social group of free-living meerkats sampled across an entire breeding season. The specific objectives of this study were to (1) validate noninvasive (fecal and urinary) gonadal steroid hormone monitoring techniques in male (testosterone) and female (estrogens, progestagens) meerkats; (2) test the feasibility of using these noninvasive methods under field conditions; (3) characterize the endocrine correlates associated with the female reproductive cycle, including estrus, gestation, and postpartum estrus; (4) examine longitudinal androgen excretion in males; and (5) determine whether social status (i.e., dominant versus subordinate) affected gonadal steroid excretion. In females, the results demonstrated the physiological validity of noninvasive monitoring in meerkats by corresponding excretory hormone concentrations to major reproductive events (i.e., estrous, pregnancy, parturition). Hormone excretory patterns during estrous intervals suggested possible mechanisms whereby reproductive suppression may operate in female meerkats. In males, androgen excretion did not correspond to changes in reproductive and aggressive behaviors, suggesting that dominance, and hence breeding access to females, was not regulated strictly by gonadal steroid production. The consistency in androgen excretion among male meerkats indicated that reproductive suppression may be mediated by behavioral (i.e., intermale aggression) rather than physiological (i.e., depressed spermatogenesis) mechanisms.

Persistent pregnanediol glucuronide secretion after gonadotrophin suppression indicates adrenal source of progesterone in premature ovarian failure.

A 2-3 fold higher urinary pregnanediol glucuronide excretion has been observed in women with premature ovarian failure, compared with age-appropriate menopausal women. Progesterone, the precursor of urinary pregnanediol glucuronide, is a secretory product of either adrenal or ovarian origin. We postulated that suppression of pituitary gonadotrophin secretion by down-regulation with a long-acting gonadotrophin-releasing hormone agonist, leuprolide acetate, would decrease ovarian but not adrenal pregnanediol glucuronide. This would demonstrate a major difference in the ovarian hormonal milieu of these two groups of women. Four volunteers with premature ovarian failure collected daily first morning voided urine samples for 1 month prior to leuprolide acetate administration. Leuprolide acetate was then administered monthly for 3 months while continuing daily urinary collection. Luteinizing hormone (LH), follicle stimulating hormone (FSH), and pregnanediol glucuronide were measured in all samples and normalized for creatinine. Comparisons of pre- and post-median values for luteinizing hormone (LH), FSH, and pregnanediol glucuronide were made using the Wilcoxon rank sum test. This demonstrated significant suppression of both LH and FSH. Pregnanediol glucuronide, however, did not demonstrate a significant decline, strongly implying an adrenal source of the enhanced excretion. The decreased pregnanediol glucuronide noted in age-appropriate menopausal women compared with premature ovarian failure is likely to be a reflection of adrenal ageing.

Urinary steroids at time of surgery in postmenopausal women with breast cancer.

Urinary steroid metabolites were measured by capillary gas chromatography in 22 postmenopausal women with operable breast cancer on day before the tumour excision and in 20 hospitalised control who were before an operation from other cause than cancer. Serum dehydroepiandrosterone-sulphat (DHEAS) and testosterone (T) level were measured by radioimmunassay in the same groups and same time. There was no significant difference in the level of urinary androgen metabolites. Pregnanediol level was significantly lower (P < 0.05) in cancer patients. In the 5 patients with positive axillary nodes the tetrahydrocortisol and alpha-cortolone levels were significantly (P < 0.05) higher than in node negative ones. There was no significant differences in the serum DHEAS and T levels. These results indicate that metabolic changes are existing in postmenopausal patients which may be a cause or a consequence of the disease.