The measurement of urinary gonadotropins for assessment and management of pubertal disorder.

OBJECTIVE: Measurement of urinary LH (uLH) and FSH (uFSH) may facilitate non-invasive pubertal assessment but there is a need for further validation by studying children and adolescents with disorders of puberty. DESIGN: 65 cases (Male: 25) with a median age of 12 years (2.9-18.1) supplied at least one non-timed urine sample for uLH and uFSH measurement by immunoassay and corrected for creatinine excretion. 25 cases were receiving GnRH-agonist (GnRH-a) at the time of sample collection. In 41 cases, urine samples were collected prior to a LHRH test and in 12 cases matched serum samples for basal LH (sLH) and FSH (sFSH) were also available. RESULTS: There was a significant correlation between sLH and uLH:uCr (r=0.82; p-value <0.001) and sFSH and uFSH:uCr (r=0.93; p-value <0.001). Based on receiver operator characteristics analysis, a uLH:uCr value of 0.05 IU/mmol as a cut-off would detect a LH peak >5U I/L with a sensitivity of 86% and a specificity of 72% with a positive predictive value of 93%. In pubertal boys (6) and girls (22) with a sLH peak >5UI/L, median uLH:uCr was 0.27 IU/mmol (0.27-0.28) and 0.17 IU/mmol (0.09-0.43), respectively. The median uFSH:uCr was 0.51 IU/mmol (0.41-0.60) for boys and 1.1 IU/mmol (0.21-2.44) for girls. In the 25 cases on GnRH-a, the median uLH:uCr for boys and girls was 0.02 IU/mmol (0.01-0.02) and 0.02 IU/mmol (0.004-0.07), respectively, and the median uFSH:uCr was 0.07 IU/mmol (0.05-0.09) and 0.27 IU/mmol (0.09-0.54), respectively. CONCLUSION: Urinary gonadotrophins reflect serum gonadotrophin concentration and may represent a reliable non-invasive method of assessing pubertal progress.

Female cancer survivors exposed to alkylating-agent chemotherapy have unique reproductive hormone profiles.

OBJECTIVE: To evaluate reproductive hormone patterns in women exposed to alkylating-agent chemotherapy. DESIGN: Prospective cohort. SETTING: University hospital. PATIENT(S): Normally menstruating mid-reproductive-age women (20-35 years old) who had previously been exposed to alkylating-agent chemotherapy for cancer treatment were compared with two healthy control populations: similarly-aged women and late-reproductive-age women (43-50 years old). INTERVENTION(S): Subjects collected daily urine samples for one cycle. MAIN OUTCOME MEASURE(S): Integrated urinary pregnanediol glucuronide (PDG) and estrone conjugate (E1c) and urinary excretion of gonadotropins (FSH and LH). RESULT(S): Thirty-eight women (13 survivors, 11 same-age control subjects, 14 late-reproductive-age control subjects) provided 1,082 urine samples. Cycle length, luteal phase length, and evidence of luteal activity were similar among the groups. As expected, ovarian reserve was impaired in cancer survivors compared with same-age control subjects but similar between survivors and late-reproductive-age control subjects. In contrast, survivors had total and peak PDG levels that were similar to same-age control subjects and higher than those observed in late-reproductive-age control subjects. Survivors had higher E1c levels than both same-age and late-reproductive-age control subjects. There was no difference in urinary gonadotropins among the groups. CONCLUSION(S): Women exposed to alkylating agents have a unique reproductive hormone milieu that is not solely explained by age or ovarian reserve. The urinary hormone profile observed in survivors appears more similar to same-age control subjects than to late-reproductive-age women with similar ovarian reserve, which may suggest that age plays a more important role than ovarian reserve in the follicular dynamics of survivors.

Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine.

Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH.

Characterization of follicle stimulating hormone profiles in normal ovulating women.

OBJECTIVE: To describe FSH profile variants. DESIGN: Observational study. SETTING: Multicenter collaborative study. PATIENT(S): A total of 107 women. INTERVENTION(S): Women collected daily first morning urine and underwent serial ovarian ultrasound. MAIN OUTCOME MEASURE(S) FSH RESULT(S): The individual FSH cyclic profiles demonstrated a significant departure from the currently accepted model. A decline in FSH levels at the end of the follicular phase was observed in only 42% of cycles. The absence of this decline was significantly associated with a shorter luteal phase and higher pregnanediol-3α-glucuronide, FSH, and LH levels at the time of ovulation. In 34% of the cycles, significant FSH variability was observed throughout the follicular phase; this variability was associated with higher body mass index and lower overall FSH and LH levels throughout the cycle. The FSH peak occurs on average 2 hours before ovulation. The FSH peak duration was shorter than the LH peak. CONCLUSION(S): These results suggest that average FSH profiles may not reflect the more complex dynamics of daily hormonal variations in the menstrual cycle. It is possible that discrepancies between the average normal FSH profile and the individual day-to-day variants can be used to detect abnormalities.

Urinary follicle-stimulating hormone as a measure of natural fertility in a community cohort.

High serum follicle-stimulating hormone (FSH) levels have been associated with diminished ovarian reserve; however, the association between high urinary FSH and reduced natural fertility has yet to be established. We sought to characterize the relationship between a single or multiple measurements of early follicular phase urinary FSH and fertility. Women (n = 209), 30 to 44 years old with no history of infertility, who had been trying to conceive for less than 3 months, provided early follicular phase urine. Participants subsequently kept a diary to record bleeding and intercourse and conducted standardized pregnancy testing for up to 6 months. A subset of women (N = 95) collected urine on cycle day 3 for up to 6 cycles. Urine was analyzed for FSH and creatinine (cr) corrected. Proportional hazard models were used to calculate fecundability ratios (FRs). Urinary FSH levels across cycles from the same woman were highly correlated (adjusted intraclass correlation = .77); within-woman variance was 3-fold lower than variance among women. Women with an initial urinary FSH level <7 mIU/mg cr exhibited a nonsignificant reduction in the probability of pregnancy (adjusted FR 0.71, 95% confidence interval [CI]: 0.45-1.13), as did women with elevated urinary FSH (≥12 mIU/mg cr; adjusted FR 0.78, 95% CI: 0.46-1.32). Using the most recent or maximum urinary FSH value did not strengthen the association. In the general population, urinary FSH levels appear to be nonlinearly associated with fertility; however, broad CIs indicate a lack of statistical significance. Repetitive testing appears to be of little benefit.

Symptom interference with work and relationships during the menopausal transition and early postmenopause: observations from the Seattle Midlife Women's Health Study.

OBJECTIVE: The aim of this study was to describe the changes in symptom interference during the menopausal transition (MT) stages and early postmenopause (PM), including the effects of age, MT-related factors (estrone, follicle-stimulating hormone, testosterone, MT stages), symptoms (hot flashes, depressed mood, awakening during the night, anxiety, backache, joint pain, forgetfulness, and difficulty concentrating), health-related factors (perceived health), and stress-related factors (perceived stress, cortisol). METHODS: A subset of Seattle Midlife Women's Health Study participants provided data during the late reproductive, early and late MT stages, or early PM (n = 184), including menstrual calendars for staging the MT; annual health reports completed between 1990 and 2008; morning urine samples assayed for estrone glucuronide, follicle-stimulating hormone, and cortisol; and symptom diary ratings several times each year. Interference was rated in the diary along with symptoms, perceived health, and stress. Multilevel modeling with an R program was used to test the patterns of symptom interference related to age, MT-related factors, symptoms, and health- and stress-related factors, with as many as 5,656 observations. Median age was 47.4 years. RESULTS: Interference with work was significantly associated with individual covariates such as perceived health, stress, hot flashes, depressed mood, anxiety, difficulty getting to sleep, awakening during the night, early morning awakening, backache, joint pain, forgetfulness, and difficulty concentrating (for hot flashes, P = 0.01; all others, P < 0.001). A final multivariate model included perceived health, stress, depressed mood, and difficulty concentrating. Interference with relationships was significantly associated with age and individual covariates such as perceived health, estrone, perceived stress, depressed mood, anxiety, sleep symptoms, backache, joint pain, forgetfulness, and difficulty concentrating (for estrone, P = 0.03; all others, P < 0.001). A final multivariate model included perceived health, stress, depressed mood, anxiety, difficulty concentrating, and awakening during the night. CONCLUSIONS: Women's reports of how much the way they felt interfered with work and relationships were influenced by both their perceived health and stress levels. Interference was also influenced by depressed mood and difficulty concentrating, suggesting that these two symptoms may be most important to address, to enhance functioning during the MT and early PM.

Absorption, distribution, metabolism and excretion of corifollitropin alfa, a recombinant hormone with a sustained follicle-stimulating activity.

The follicle-stimulating hormone (FSH) analog corifollitropin alfa (Org 36286) is a potent FSH receptor agonist with an extended plasma half-life due to the fusion of the carboxy-terminal peptide (CTP) of the human chorionic gonadotropin beta-subunit and the human FSH beta-subunit. The absorption, tissue distribution, metabolism and excretion of corifollitropin alfa were studied in rats following a single subcutaneous administration of [(125)I]corifollitropin alfa. The biological activity of [(125)I]corifollitropin alfa was confirmed by an in vitro FSH receptor transactivation assay. Radioactivity in blood, serum, tissues and excreta was determined by radiometry up to 168 h post dosage. A drug-specific distribution occurred mainly to ovaries and the renal system. The distribution was similar in albino and pigmented rats, ruling out effects of melanin binding on distribution. Metabolites were studied in urine and serum by SDS-PAGE. The maximum concentration of 12 h indicated a slow absorption and excretion. Radioactivity was mainly (86%) excreted via urine. 90% of the radioactivity in serum was identified as [(125)I]corifollitropin alfa, whereas only 7-15% of the radioactivity in urine was identified as [(125)I]corifollitropin alfa and its dissociation products, the alpha- and beta-subunits (including its CTP part). The remainder of the radioactivity in either matrix represented low-molecular-weight compounds resulting from catabolism and deiodination. In conclusion, the metabolic fate of corifollitropin alfa strongly resembles that of endogenous glycoprotein hormones, which predominantly consists of kidney clearance and the urinary excretion of the intact protein in parallel to kidney catabolism.

Well-being during the menopausal transition and early postmenopause: a within-stage analysis.

PURPOSE: To identify whether menopausal transition (MT)-related factors--including MT stage, hot flash severity, levels of estrone glucuronide (E1G) and follicle stimulating hormone (FSH); number of negative life events; or personal resources of mastery and social support--are associated with stage specific well-being. METHODS: Women from the Seattle Midlife Women's Health Study (N=334) provided at least one annual health questionnaire and a menstrual calendar; a subset provided first morning voided urine specimens assayed for E1G and FSH. Descriptive statistics were calculated and Pearson's product-moment correlations were estimated. RESULTS: Mean levels of well-being were the same across MT stages (4.1, range 1.8-5.9). Physiologic variables were not significantly correlated with well-being: E1G levels (r = - .11 to 0.16), FSH levels (r = - .17 to .20) and hot flash severity (r = - .07 to .05). Significant correlations were observed between well-being and number of negative life events (r = - .48 to - .33, p < or = 0.01), mastery (r = .51-.64, p < or = 0.01), and satisfaction with social support (r = .04 non-significant) to r =.41, (p < or = 0.01). CONCLUSION: The association of well-being with life events and personal resources, and not MT-related indicators, hormone levels or vasomotor symptoms, supports work by other researchers. Further study is needed to determine whether patterns of well-being vary across the MT, and if so, what might predict that variability.

Hot flash severity in hormone therapy users/nonusers across the menopausal transition.

OBJECTIVES: The purpose of this study was to examine the pattern of and factors that influence hot flash severity across the menopausal transition (MT) and early postmenopause (PM). METHODS: Women from the Seattle Midlife Women's Health Study (N=302) provided data for these analyses: at least one annual health questionnaire and a menstrual calendar. A subset of women provided a first morning voided urine specimen from 1997 through 2005. Urine samples were assayed for estrone glucuronide and FSH. Linear mixed effects modeling was used to identify change in hot flash severity scores over time, including the relationship to age, MT-related, psychosocial and lifestyle factors. RESULTS: Increases in hot flash severity were associated with late transition stage, early postmenopause, use of HRT, duration of early transition stage, age of entry into early PM and level of FSH. Age of entry into early transition and estrone levels were associated with decreased hot flash severity. Not associated with hot flash severity were being in early transition stage, age of entry into or duration of late transition stage and all of the psychosocial (anxiety, stress, depressed mood) and lifestyle variables (BMI, activity level, sleep, alcohol use). CONCLUSIONS: Variables associated with reproductive aging independently predicted changes in hot flash severity; psychosocial and lifestyle variables did not. The effect of age dropped out when factors associated with reproductive aging were considered. Use of HRT ameliorated but did not eliminate severe hot flashes suggesting that there is room for alternative approaches less likely to cause harm.

Symptoms during the menopausal transition and early postmenopause and their relation to endocrine levels over time: observations from the Seattle Midlife Women's Health Study.

OBJECTIVE: To determine whether hot flashes, depressed mood, sleep, cognitive and sexual symptoms correlate with urinary follicle-stimulating hormone (FSH), estrone (E(1)G), and testosterone (T) and with each other during the menopausal transition and early postmenopause (PM). METHODS: Forty-one women who transitioned from middle or late transition stage to PM rated symptoms and provided monthly urine specimens as part of a longitudinal study of the menopausal transition. RESULTS: Correlations between endocrine levels and symptom severity ratings over time revealed that hot flash severity was significantly and positively related to FSH and negatively to E1 G. Vaginal dryness was positively correlated with FSH and negatively correlated with T. Decreased sexual desire was correlated negatively with E(1)G levels. Forgetfulness was positively correlated with FSH; difficulty concentrating was negatively correlated with T. Severity of sleep symptoms and depressed mood were not correlated with E(1)G, FSH, or T. Correlations among the symptoms revealed that severity of hot flashes was associated with sleep disruption and forgetfulness. Depressed mood was correlated with sleep disruption, difficulty concentrating, and decreased sexual desire but not with hot flashes or vaginal dryness. Awakening during the night was correlated with decreased sexual desire and vaginal dryness, as well as hot flashes. Forgetfulness was associated with hot flashes and difficulty concentrating, whereas difficulty concentrating was associated with depressed mood and early awakening. CONCLUSIONS: Symptoms many women experience during the menopausal transition and early PM are related to different endocrine levels (FSH, E(1)G, and T).

Embryo quality based on ovulation induction: defining the differences.

Patients undergoing IVF, with or without intracytoplasmic sperm injection, were treated with either recombinant human FSH or urine-derived FSH. Response to ovarian stimulation was monitored by ultrasound examinations and measurement of serum oestradiol concentrations. To define any differences in embryo quality and hence assisted reproductive technology success rates, a retrospective analysis of 811 recombinant FSH versus 555 urinary FSH cycles was undertaken. Embryo quality was assessed as embryo cell number and degree of fragmentation. Implantation and ongoing pregnancy rates were also compared. Use of recombinant FSH resulted in a higher percentage of mature oocytes, improved embryo cleavage, with more embryos available for freezing and higher implantation rates compared with urinary FSH. Oocyte and embryo quality were superior when recombinant FSH was used for ovarian stimulation compared with urinary FSH.

Impact of highly purified urinary FSH and recombinant FSH on haemostasis: an open-label, randomized, controlled trial.

BACKGROUND: It has recently been suggested that recombinant FSH administration may result in an increased risk of venous thrombosis. An open-label, randomized, controlled trial was carried out to compare the impact of urinary and recombinant FSH on haemostasis. METHODS: Fifty infertile women were randomized, using a random number generator on a personal computer, to receive either highly purified urinary FSH (u-hFSH) or recombinant human FSH (r-hFSH); a starting dose of 150 IU. Human chorionic gonadotrophin 10000 IU was administered once there was at least one follicle > or =18 mm. The luteal phase was supported with progesterone 50 mg/day for at least 15 days. Fifty normally menstruating women were recruited as controls. Repeated measurements of estradiol, progesterone, prothrombin time (PT) expressed as INR, activated partial thromboplastin time (APTT) ratio, fibrinogen (FBG), factor VIII (FVIII), normalized activated protein C ratio (nAPC ratio), antithrombin III activity (AT), protein C activity (PC), protein S activity (PS), tissue-type plasminogen activator antigen (t-PA), type 1 plasminogen activator inhibitor (PAI), prothrombin fragments 1+2 (F1+2), were performed during both hyperstimulated and natural cycles, and at onset of the following menstruation or at 8 weeks of pregnancy. RESULTS: At the end of gonadotrophin administration PT INR increased in the u-hFSH group, while AT and t-PA significantly decreased. In the patients treated with r-hFSH, only F1+2 significantly decreased. No significant changes were observed in the control group. In the luteal phase FBG increased significantly in all groups. In the u-hFSH group no other significant changes were noted compared to pre-ovulatory values, while compared to baseline values AT, PS and t-PA significantly decreased. In the r-hFSH group during the luteal phase PT INR significantly decreased, but did not differ from baseline levels. Other parameters such as FBG, FVIII, t-PA, rose significantly, but only FVIII and FBG values were significantly higher than baseline levels. In the women who became pregnant a significant increase in t-PA and a significant decrease in PS at the midluteal phase were observed. After one month all the haemostatic parameters returned to baseline value if pregnancy failed to occur, while in the pregnant women a significant increase in FVIII and a significant decrease in PS were observed. CONCLUSIONS: Ovarian stimulation with recombinant FSH does not influence coagulation and fibrinolysis significantly, as already reported for urinary gonadotrophins. The moderate changes induced by both treatments are no longer detectable after 4 weeks.