Urinary LH Research Articles

Follicular diameter and hCG administration do not affect pregnancy rates after clomiphene citrate and intrauterine insemination

Objective: Success rates of clomiphene citrate (CC) with timed intrauterine insemination (IUI) in treating infertile couples vary widely in the literature. The heterogeneity of the populations studied as well as differences in the timing of the insemination with regard to follicular size and the preovulatory LH rise and/or the administration of human chorionic gonadotropin (hCG) may contribute to this variance. The intent of this study was to evaluate the pregnancy rates after stimulation with CC/IUI with respect to follicular diameter.Design: Retrospective analysis.Material and Methods: Four hundred ninety-two women who underwent 691 cycles of ovarian stimulation with CC/IUI at the Johns Hopkins Fertility Center between 1/1/98 and 12/30/01 were included in the study. All patients received 50mg/day of CC for 5 days beginning on the 5th day of their menstrual cycle or five days after progesterone-induced withdrawal bleeding. A single IUI was performed in all cases either 24 hours following a spontaneous serum LH surge (>25mIU/ml) or 36 hours after intra-muscular hCG administration (10,000 IU) if the largest follicle had reached a diameter of at least 17mm and the LH was less than 25mIU/ml. For statistical evaluation, t-test, chi square, and correlation analysis were used.Results: There were 223 cycles with a spontaneous LH surge and 468 cycles in which hCG was administered. There were no differences between the two groups in mean age or indication for treatment. The pregnancy rates for the LH surge group (12.4%) and the hCG group (14%) were not statistically different. A spontaneous LH surge was observed at follicular diameters ranging from 12 to 35mm. In the hCG group, the diameter of the leading follicle measured between 17 and 36mm at the time of hCG administration. There was no correlation between follicular diameter and pregnancy outcome in either group. Conclusion: IUI following ovarian stimulation with CC is equally effective 24 hours after a spontaneous LH surge or 36 hours after administration of hCG. Spontaneous LH surges were observed at a variety of follicular sizes with comparable pregnancy rates. Follicular diameter at the time of hCG administration does not appear to have a major impact on pregnancy outcome. Sonographic monitoring of patients treated with CC/IUI did not appear to offer any additional benefit. A urinary LH detection kit may be a more cost-effective way to predict ovulation and time IUI accordingly. Objective: Success rates of clomiphene citrate (CC) with timed intrauterine insemination (IUI) in treating infertile couples vary widely in the literature. The heterogeneity of the populations studied as well as differences in the timing of the insemination with regard to follicular size and the preovulatory LH rise and/or the administration of human chorionic gonadotropin (hCG) may contribute to this variance. The intent of this study was to evaluate the pregnancy rates after stimulation with CC/IUI with respect to follicular diameter. Design: Retrospective analysis. Material and Methods: Four hundred ninety-two women who underwent 691 cycles of ovarian stimulation with CC/IUI at the Johns Hopkins Fertility Center between 1/1/98 and 12/30/01 were included in the study. All patients received 50mg/day of CC for 5 days beginning on the 5th day of their menstrual cycle or five days after progesterone-induced withdrawal bleeding. A single IUI was performed in all cases either 24 hours following a spontaneous serum LH surge (>25mIU/ml) or 36 hours after intra-muscular hCG administration (10,000 IU) if the largest follicle had reached a diameter of at least 17mm and the LH was less than 25mIU/ml. For statistical evaluation, t-test, chi square, and correlation analysis were used. Results: There were 223 cycles with a spontaneous LH surge and 468 cycles in which hCG was administered. There were no differences between the two groups in mean age or indication for treatment. The pregnancy rates for the LH surge group (12.4%) and the hCG group (14%) were not statistically different. A spontaneous LH surge was observed at follicular diameters ranging from 12 to 35mm. In the hCG group, the diameter of the leading follicle measured between 17 and 36mm at the time of hCG administration. There was no correlation between follicular diameter and pregnancy outcome in either group. Conclusion: IUI following ovarian stimulation with CC is equally effective 24 hours after a spontaneous LH surge or 36 hours after administration of hCG. Spontaneous LH surges were observed at a variety of follicular sizes with comparable pregnancy rates. Follicular diameter at the time of hCG administration does not appear to have a major impact on pregnancy outcome. Sonographic monitoring of patients treated with CC/IUI did not appear to offer any additional benefit. A urinary LH detection kit may be a more cost-effective way to predict ovulation and time IUI accordingly.

Diminished ovarian reserve in female childhood cancer survivors with regular menstrual cycles and basal FSH

Approximately 70% of children treated for cancer are cured, however the treatment often impairs reproductive function. The aim of the present study was to assess ovarian reserve in women with an apparently normal ovarian function, who were cured of cancer in childhood. Twenty-one female survivors with regular menstrual cycles and basal FSH <10 IU/l were included. The control group included 21 healthy age-matched regularly cycling women. On cycle day 2-5, ovarian volume and number of small antral follicles (2-5 mm) were evaluated with transvaginal ultrasonography. Repeated sonography, urinary LH testing, and endocrine assessments were performed during the actual cycle. Cycle length was calculated. The female survivors had significantly smaller volume per ovary (4.9 versus 6.8 cm(3); P < 0.01), a lower number of small antral follicles per ovary (4.5 versus 8.0; P < 0.01), and lower total number of follicles per ovary (8 versus 11, P < 0.01). Hormonal profiles were similar, but the mean cycle length of the female survivors was significantly shorter than in the control group (28.3 versus 31.0 days; P < 0.05). Childhood cancer survivors with regular menstrual cycles and basal FSH <10 IU/l seem to have a diminished ovarian reserve. Consequently, they may have a shortened reproductive span and an early menopause.

A pharmacokinetic and endocrine comparison of recombinant follicle-stimulating hormone and human menopausal gonadotrophin in polycystic ovary syndrome

Elevated LH concentrations are frequently encountered in patients with polycystic ovary syndrome (PCOS) and increased LH (either endogenous or superimposed through the use of HMG) may have detrimental effects on reproductive function. In spite of this, FSH-only products and HMG have been used indiscriminately for ovulation induction – on the basis that the administration of HMG to patients with PCOS, who are not receiving GnRH agonists, does not result in significant increases in serum LH concentrations as judged by daily single blood samples. However, both endogenous and exogenous LH have a relatively short terminal half-life and studies have reported normal serum LH, but abnormal urinary LH and emphasized that early morning urinary measurements are more informative than those in serum because they reflect nocturnal LH secretion. Therefore, the present study was undertaken to perform a pharmacokinetic and endocrine comparison of recombinant human FSH and HMG in PCOS patients including LH measurements in the urine. Five PCOS patients receiving s.c. recombinant human FSH (rhFSH) and five PCOS patients receiving i.m. HMG for ovulation induction according to a chronic low-dose step-up regimen underwent blood and urine sampling at the following study points: Point 0 was the day of HCG injection; Points 1 to 5 corresponded to days HCG −1 to −2; −3 to −4; −5 to −6; −7 to −8; and −9 to −10; respectively. Serum hormone measurements included oestradiol, FSH, LH, progesterone, inhibin A, androstenedione, testosterone, and free testosterone index. FSH and LH were also measured daily in 8-h urine samples reflecting overnight renal urine secretion. Hormone concentrations calculated as the area under the curve showed that both FSH and LH concentrations in urine were significantly higher in HMG group than in group rhFSH. It is concluded that both LH and FSH concentrations significantly accumulate in the urine of PCOS patients receiving HMG for ovulation induction in a chronic low-dose protocol as compared with rhFSH treatment.

Mifepristone-induced vaginal bleeding is associated with increased immunostaining for cyclooxygenase-2 and decrease in prostaglandin dehydrogenase in luteal phase endometrium.

The mechanism of mifepristone-induced vaginal bleeding and endometrial shedding was investigated in 13 women who took 200 mg mifepristone in the midluteal phase on d 8 after the onset of the urinary LH surge (LH+8). Endometrial biopsies were collected, 6-24 h after mifepristone (group 1, n = 7) or 36-48 h after mifepristone (group 2, n = 6), and compared with those from a control group in the midluteal phase (n = 7). All women reported vaginal bleeding commencing 36-48 h after taking mifepristone. Treatment with mifepristone significantly reduced serum progesterone levels in all women, when compared with the controls (13.2 nM vs. 34.8 nM, P = 0.001). After mifepristone, a significant increase in cyclooxygenase-2 immunoreactivity was apparent at 36-48 h (P = 0.0018), whereas prostaglandin 15 dehydrogenase enzyme-positive immunostaining declined, to be virtually absent by 36-48 h in both glands and in stroma (P < 0.05). There was no change in intensity or distribution of staining for steroid receptors after mifepristone. The changes in immunostaining for cyclooxygenase-2 and prostaglandin 15 dehydrogenase strongly support the hypothesis that an increase in the local concentration of prostaglandins in the endometrium is involved in the mechanism of bleeding induced by mifepristone in the luteal phase.

Endometrial dating revisited: a randomized systematic study of secretory phase histologic characteristics in normally cycling fertile women

Objective: More than five decades have elapsed since publication of the classical work of Noyes, Hertig, and Rock that established histologic criteria for dating the endometrium. In more recent years, the accuracy, reliability, and clinical utility of those criteria have been seriously challenged. To address the existing controversies and to more rigorously define the histologic characteristics of the normal secretory phase endometrium, we studied a group of healthy, normally cycling, proven fertile women. Design: Randomized observational study. Materials/Methods: A total of 130 volunteers (age 20–35 yr) were recruited to participate. All reported regular, spontaneous menses (intermenstrual interval 25–35 d) and had at least one liveborn child and a normal uterus as observed by transvaginal ultrasound examination. Each subject monitored urinary LH excretion daily from cycle day 10 until detection of the midcycle surge (luteal day 0) and was randomized to return 1–14 days thereafter for endometrial sampling and measurement of serum estradiol and progesterone concentrations. After processing, adjacent sections of each tissue specimen were examined by three independent gynecologic pathologists masked to the day of sampling and scored for 32 different histologic features that were defined prior to initiation of the study. For each histologic feature, mean scores and the extent of between tissue and inter-observer variation were determined for each luteal day (1–14). Features were ranked according to the strength of inter-specimen correlation with luteal day, score change over time, and inter-observer variation. A final rank order was determined based on consideration of all three performance measures. Results: A total of 121 tissue specimens were adequate for evaluation. Our observations were in many ways consistent with the classical description, but in others, very different. Whereas some of the traditional histologic dating criteria were among the features exhibiting the best overall performance in our objective analysis, others clearly were not. Inter-observer and inter-specimen variation for the best performing features were greatest during the early secretory phase and the variation decreased as luteal day increased. The most dramatic, consistent, and therefore discriminating changes in score for the best performing features were observed during the putative implantation window (luteal days 6–10). Conclusions: The characteristics of secretory phase endometrial histology exhibit greater individual variations than indicated in the classical description. We were unable to identify any one or combination of histologic features that could effectively define a specific luteal day. Nevertheless, carefully timed endometrial sampling during the implantation window may yet provide a means to objectively confirm normal endometrial maturation. Supported by: University of North Carolina, Division of Reproductive Endocrinology and Infertility.

Urinary follicle-stimulating hormone peak as a biomarker for estimating the day of ovulation

Objective: To evaluate urinary follicle-stimulating hormone (FSH) as a biomarker for the day of ovulation. Design: Prospective observational study. Setting: Clinical research center. Patient(s): Thirteen women were monitored with measurements of serum and urinary hormones and ovarian ultrasonography during 20 menstrual cycles. Data on urinary hormones and ultrasound evaluations from a total of 65 menstrual cycles from 42 women were analyzed. Intervention(s): Blood and/or urine samples were collected daily. Daily transvaginal ultrasonography was used to detect follicular collapse. Main Outcome Measure(s): LH, FSH, and E 2 were measured in serum. FSH, estrone conjugates (E1C), and pregnanediol-3-glucuronide (PdG) were analyzed in urine. The day of luteal transition (DLT) was calculated using two algorithms. Result(s): In 20 cycles, the urinary FSH peak was closer to the day of follicular collapse (−0.85 days) than was the peak day of serum E 2 and the day of luteal transition, as calculated by one algorithm. The FSH peak was not closer to the day of follicular collapse than the peak values of urinary LH, serum FSH, or the day of luteal transition as calculated by a second algorithm. The most consistent correspondence between a hormone peak and ovulation was for serum E 2, serum FSH, serum LH, and urinary FSH. In 65 cycles for which urinary hormone data and ultrasound evaluations were available, the urinary FSH peak occurred within 1 day of follicular collapse in 97% of cycles. Conclusion(s): Urinary FSH is a useful biomarker for estimating the day of ovulation in population-based studies.

The Clinical Usefulness of Salivary Progesterone Measurement for the Evaluation of the Corpus luteum Function

The present study was designed to construct reliable daily salivary progesterone profiles throughout the luteal phase to accurately evaluate the corpus luteum function. Furthermore, we investigated the clinical relevance of a simple midluteal salivary progesterone estimation for the diagnosis of luteal phase insufficiency by determining the diagnostic efficiency and cutoff values. A total of 121 women were divided into 3 groups; normal luteal function, luteal phase insufficiency and unclassified group, based on basal body temperature recordings and serum progesterone levels at 3 sampling points during the midluteal phase. Salivary progesterone values across the luteal phase of the normal luteal function group were significantly increased from day 1 to day 4, remained constant from day 5 to day 9 (mean ± SD, 318 ± 170 pmol/l on day 5, 287 ± 169 pmol/l on day 9; urinary LH surge = day 0) and decreased thereafter. Salivary progesterone concentrations in the luteal phase insufficiency group showed significantly lower values compared with those in the normal group between days 3 and 10. The cutoff values of 189 pmol/l in the midluteal phase yielded a sensitivity of 78.0% and a specificity of 76.5%. Our results suggest that daily salivary progesterone profiles during the luteal phase and a simple estimation of midluteal salivary progesterone appeared to be useful for the diagnosis of luteal phase defects.

The effect of nonsteroidal anti-inflammatory drugs on ovulation: a prospective, randomized clinical trial

Objective: To assess the effect of ibuprofen, a nonspecific inhibitor of prostaglandin synthesis, on ovulation. Design: Prospective, randomized, double-blind, placebo-controlled cross-over study. Setting: University Medical Center. Patient(s): Twelve normally cycling women between ages 20 and 40. Intervention(s): Subjects were randomized to either oral ibuprofen (800 mg) or placebo three times per day, beginning when the maximum diameter of the leading follicle reached 16 mm by ultrasound, and continuing for 10 days total. The second cycle was a washout period, and in the third cycle, the subjects were crossed over to the alternate regimen from the first cycle. The probability of delayed follicular collapse was determined using the binomial distribution, and changes in P levels were compared using the paired t test. Main Outcome Measure(s): Urinary LH surge, follicular collapse by serial transvaginal ultrasonography, and serum midluteal P levels. Result(s): Eleven of 12 subjects detected an LH surge with both ibuprofen and placebo. Five of 11 women demonstrated a ≥2-day increase in time interval from detection of the LH surge to follicular collapse, and 3 of those 5 had been randomized to ibuprofen. This represents a 27% (3 of 11; 95% confidence limits: 1%, 53%) rate of delay for follicular collapse for ibuprofen. There was no difference in average midluteal P levels for ibuprofen or placebo. Conclusion(s): If ibuprofen inhibits follicular collapse, this effect is seen in a small group of study subjects, and this information should be clinically reassuring to patients who take nonsteroidal anti-inflammatory drugs. Serum midluteal P levels were unaffected by administration of ibuprofen.

Metabolism of gonadotropins: comparisons of the primary structures of the human pituitary and urinary LH beta cores and the chimpanzee CG beta core demonstrate universality of core production.

The gonadotropins are a family of closely related heterodimeric glycoprotein hormones homologous in structure to disulfide-knot growth factors. Metabolic proteolytic processing in vivo of this disulfide cross-linked region results in urinary excretion of a residual highly stable core structure. The primary structure of the pituitary form of the hLH beta core was reported earlier, but it has proved difficult to isolate the urinary core, although antibodies to the pituitary core demonstrated its presence. By conventional and immunoaffinity methods, the urinary core has been isolated and its structure determined by both chemical and mass spectrometric methods. The urinary hLH beta core is the same as the pituitary-extracted hLH beta core, beta 6-40 disulfide bridged to beta 55-93, except that the pituitary core is more heterogeneous containing also beta 49-93. These findings imply a dual origin of urinary cores, both directly from a secreting tissue and by kidney processing of circulating hormone. We also found that pregnant chimpanzees excrete a CG beta core with a primary structure identical to that of the human CG beta core of pregnancy. In conclusion, gonadotropin core generation and urinary excretion of nearly identical gonadotropin metabolites is common among primates. Although possible biological functions of these core fragments remain unproven, they have diagnostic utility because of their stability and abundance.

Hypothalamic-pituitary-ovarian dysfunction after prepubertal chemotherapy and cranial irradiation for acute leukaemia.

We assessed adult hypothalamic-pituitary-ovarian function following treatment with chemotherapy and cranial irradiation for childhood acute lymphoblastic leukaemia. The patients (n = 12) had median age at diagnosis of 4.7 years, and at assessment of 20.8 years. They collected a daily urine sample over two to five consecutive menstrual cycles (total of 41 cycles) for analysis of LH and steroid excretion. Blood sampling and ovarian ultrasound examination was performed in the early follicular phase. Sixteen healthy women with regular menstrual cycles were recruited as controls. Urinary LH excretion was significantly lower in patients throughout the cycle, particularly during the LH surge (P < 0.0001). The length of the luteal phase was significantly shorter in patients than in normal controls (12.2 +/- 0.3 versus 13.6 +/- 0.4 days, P = 0.01) with a high prevalence of short (< or =11 days) luteal phases (15/39 cycles). Luteal phase pregnanediol excretion was slightly but not significantly lower. Follicular and luteal phase excretion of oestrone was lower in patients than in controls (P = 0.01). Early follicular phase plasma oestradiol was also lower in the patient group (P = 0.032) although LH, FSH, inhibin A and B concentrations were similar. These data indicate that treatment for childhood leukaemia results in subtle ovulatory disorder in some patients, probably related to cranial irradiation. Follow-up of these women is required to detect any effect on reproductive potential.

Comparison of several one-step home urinary luteinizing hormone detection test kits to OvuQuick®

Objective: To compare clinical accuracy and ease of use for several of the new rapid one-step home urinary LH detection kits compared with the preexisting OvuQuick brand LH detection kit (designated as the standard).Design: Prospective cohort study.Setting: University-based infertility clinic.Patient(s): All women undergoing intrauterine inseminations at the clinic, regardless of infertility diagnosis, were offered enrollment during a 28-month study period.Intervention(s): Each participant was supplied three one-step test kits (OvuQuick One-Step, ClearPlan Easy, and SureStep) in addition to a multistep OvuQuick test kit and instructed to run the tests in parallel on the same urine sample and to record the results. Urine testing was performed every 12 hours, beginning 3 days before the anticipated onset of an LH surge, and continued with the one-step kits every 12 hours for 48 hours after the surge was first detected by OvuQuick. Subjects also completed questionnaires evaluating the use of each test kit.Main Outcome Measure(s): Correlation of LH surge detection by one-step kits in comparison to surge detection by OvuQuick. Satisfaction and ease of use questionnaires.Result(s): Sixty-three volunteers returned useable data, from which 81 evaluable cycles were analyzed. The majority of patients found the newer one-step kits to be easier to use and less time consuming than OvuQuick. The three one-step kits detected the LH surge within ± one testing period (±12 hours) of detection by OvuQuick 68%–84% of the time. OvuQuick One-Step, with modified instructions allowing for an equal color intensity in the test and reference areas, had the highest correlation with OvuQuick (84%). However, with a study power (α = 0.05, β = 0.10) sufficient to detect a 10% difference between Ovuquick and each one-step kit, all one-step kits were statistically equivalent to each other. There was no cycle in which a one-step kit detected a positive LH surge but OvuQuick did not.Conclusion(s): One-step urinary LH kits are easier for patients to use than a multistep home urinary LH kit and have reasonable correlation with the multistep kit when used clinically for timing artificial inseminations.

Reliability of Ovulation Tests in Infertile Women

In Brief Objective To assess the reliability of the most widely used clinical methods for predicting or confirming ovulation. Methods We monitored spontaneous cycles in 101 infertile women using basal body temperature (BBT), transvaginal ultrasound, a urinary stick system for LH surge, and three serum progesterone measurements in the midluteal phase. Transvaginal ultrasound monitoring was standard for ovulation detection and sensitivity. We calculated specificity and accuracy of each method compared with that standard. Results Follicular development and ultrasound evidence of ovulation were confirmed in 97 of 101 cycles (96%). Urinary LH surge preceded follicular rupture assessed by ultrasonography in all cycles and showed concordance with ultrasound-evidenced ovulation in 98 of 101 cases. The timing of BBT nadir had wide variability, and BBT and ultrasonography agreed in a similar percentage of cases (74%). Midluteal serum progesterone assessments showed ovulatory values in 93 subjects, and ovulation was concordant with ultrasonography in 90 subjects. Conclusion Urinary LH was accurate in predicting ovulation with ultrasonography as the standard for detection, but time varied widely. The nadir of BBT predicted ovulation poorly. The BBT chart was less accurate for confirming ovulation, whereas a single serum progesterone assessment in midluteal phase seemed as effective as repeated serum progesterone measures. Ultrasonography, urinary LH surge detection, and serum progesterone assessment seem to be the most effective methods for predicting and confirming ovulation.

Reliability of ovulation tests in infertile women

Objective: To assess the reliability of the most widely used clinical methods for predicting or confirming ovulation. Methods: We monitored spontaneous cycles in 101 infertile women using basal body temperature (BBT), transvaginal ultrasound, a urinary stick system for LH surge, and three serum progesterone measurements in the midluteal phase. Transvaginal ultrasound monitoring was standard for ovulation detection and sensitivity. We calculated specificity and accuracy of each method compared with that standard. Results: Follicular development and ultrasound evidence of ovulation were confirmed in 97 of 101 cycles (96%). Urinary LH surge preceded follicular rupture assessed by ultrasonography in all cycles and showed concordance with ultrasound-evidenced ovulation in 98 of 101 cases. The timing of BBT nadir had wide variability, and BBT and ultrasonography agreed in a similar percentage of cases (74%). Midluteal serum progesterone assessments showed ovulatory values in 93 subjects, and ovulation was concordant with ultrasonography in 90 subjects. Conclusion: Urinary LH was accurate in predicting ovulation with ultrasonography as the standard for detection, but time varied widely. The nadir of BBT predicted ovulation poorly. The BBT chart was less accurate for confirming ovulation, whereas a single serum progesterone assessment in midluteal phase seemed as effective as repeated serum progesterone measures.

Luteal progesterone relates to histological endometrial maturation in fertile women.

To examine the relationship between endometrial histological maturation and reproductive hormones, we studied 11 fertile women, aged 18-37 yr. All participants had had at least 1 previous pregnancy and cycled regularly, every 25-35 days. Women collected daily, first morning voided urine for measurement of estradiol and progesterone metabolite excretion, estrone conjugates (E1c), and pregnanediol glucuronide (Pdg), respectively, throughout the cycle of study. Hormones were normalized for creatinine. Between 7-9 days after home detection of a LH surge (Sure Step), participants underwent an endometrial biopsy using a small bore (Pipelle) catheter. Tissue was prepared for histological and biochemical analyses. The histological analysis is reported herein. Endometrium was dated by 3 authors (N.S., D.H., and S.P.), all of whom were blinded to the participant's identity or timing of biopsy within her cycle. Final dating was agreed upon based upon the method of Noyes et al. E1c and Pdg were integrated throughout the cycle using the trapezoidal rule, and correlations were sought between deviation from expected histology (based upon urinary hormones and LH surge) and integrated hormone values. E1c varied over a 2-fold range in these normal women, from 1196-2040 ng/cycle. Pdg excretion was much more variable, ranging from 22-119 microg/cycle. No relationship could be found between histological lagging of endometrial maturation and lower excretion of E1c. A moderate correlation was observed (Spearman's r = 0.6; P < 0.05) between degree of histological maturation and integrated Pdg. Of two women with evidence of a disparity between gland and stromal development (glands lagging behind stroma by >2 days), one excreted 24 microg Pdg/cycle, the next to lowest value. We conclude that normal fertile women experience a wide range of hormone concentrations in the face of normal endometrial maturation. Progesterone appears to exert a dose-related effect on endometrial maturation, and the techniques we used, although relatively crude clinical measures, appeared to be sufficient to detect this relationship.

Results of In Vitro Fertilization (IVF) and Intracytoplasmic Sperm Injection (ICSI) with Single Oocyte in Unstimulated Cycles

Results of In Vitro Fertilization (IVF) and Intracytoplasmic Sperm Injection (ICSI) with Single Oocyte in Unstimulated Cycles

Gonadal Source of Testosterone Metabolites in Urine of Male Cotton-Top Tamarin Monkeys (Saguinus oedipus)

Examining gonadal function in the small excitable cotton-top tamarin monkey (Saguinus oedipus) requires noninvasive sampling techniques. Two studies were performed to identify the quantifiable urinary metabolites of testosterone in cotton-top tamarins and which of the measurable metabolites would best reflect a gonadal source of testosterone secretion. In the first study, we injected unlabeled testosterone i.m. in males at either 500-ng or 1-μg levels. Urine samples were analyzed for androgens and estrogens. Testosterone and dihydrotestosterone (DHT) increased significantly following the injections in test males but not in control males. No significant increases in androstenedione occurred. Mean levels of estradiol and estrone did not consistently increase during the 5 days following injection. In the second study, a gonadotropin-releasing hormone antagonist, Antide, was used to block LH stimulation of gonadal steroidogenesis. Males given Antide at either a 6 mg/kg dose or an 18 mg/kg dose showed significantly lower levels of urinary LH than controls. At the higher Antide dose, testosterone levels were significantly reduced during weeks 1 and 2 posttreatment, whereas DHT levels significantly declined during the 2nd week posttreatment. Estradiol levels were highly variable prior to treatment but decreased significantly following treatment, whereas estrone levels remained variable throughout. These results indicate that measurement of urinary testosterone and possibly DHT reflect gonadal function in male cotton-top tamarins. Other sources of urinary estrogens may occur for the male cotton-top tamarin, but these data suggest that a substantial part of urinary estradiol is from gonadal sources, whereas urinary estrone appears to be mainly from extragonadal sources.

Use of integrins to date the endometrium

Objective: To compare traditional histologic dating criteria of the endometrium with immunohistochemical criteria based on epithelial integrin expression during the menstrual cycle. Design: Prospective clinical study. Setting: An academic teaching hospital. Patient(s): Fertile and infertile women undergoing endometrial biopsy. Main Outcome Measure(s): Immunohistochemical staining intensity and distribution (HSCORE) of three integrins and traditional histologic endometrial dating. Result(s): In 1,501 endometrial specimens, phase assignment-based integrin staining was 95% and 85% concordant with histology for the proliferative and early secretory phase, respectively, but only 54% and 49% concordant for the middle and late secretory phase, respectively. The greatest disagreement occurred during the midluteal phase. Of 1,090 patients who underwent sampling 6–10 days after detection of a urinary LH surge (corresponding to cycle days 20–24), multiple logistic regression analysis revealed that endometriosis was positively correlated and male factor infertility was negatively correlated with absent β3 integrin subunit expression. Diagnosis and absent epithelial α4β1 expression were not related. Patient age was not correlated with the incidence of abnormalities in integrin expression. Conclusion(s): Traditional histologic dating of the endometrium has remained the gold standard for nearly 50 years. Although the use of marker proteins provides additional information and may reflect endometrial function or receptivity, such markers cannot yet replace traditional methods of endometrial assessment.

Efficacy of methods for determining ovulation in a natural family planning program

Objective: To evaluate the efficacy in ovulation detection of methods used in natural family planning in comparison with pelvic ultrasonography. Design: Prospective analysis of ovulation detection by natural family planning methods and ultrasonography. Setting: Natural family planning clinic, Department of Obstetrics and Gynecology, University of Naples “Federico II”. Patient(s): Forty healthy women who were highly motivated to use natural family planning. Intervention(s): None. Main Outcome Measure(s): Transvaginal ultrasonographic findings, urinary LH levels, salivary β-glucuronidase activity, salivary ferning levels and characteristics of cervical mucus, and BBT. Result(s): Urinary LH level determination yielded a 100% correlation with the simultaneous ultrasonographic diagnosis of ovulation. Mucus sensations and characteristics yielded a 48.3% correlation when simultaneously evaluated with ovulation. β-Glucuronidase levels yielded a 27.7% correlation. The salivary ferning test had a 36.8% ovulation-detection rate the day of ovulation, but 58.7% of results were uninterpretable. Body temperature measurements yielded a 30.4% correlation with the simultaneous ultrasonographic diagnosis of ovulation. Conclusion(s): Measuring urinary LH levels is an excellent method for determining ovulation. Although variations in mucus characteristics and basal body temperature correlate somewhat with ovulation, the length of the fertile period is overestimated with these methods. The salivary ferning test and measurement of β-glucuronidase levels are not good methods for home ovulation testing.

Anovulation after precocious pubarche: early markers and time course in adolescence.

Adolescent girls with a history of precocious pubarche (PP) are known to be at increased risk for ovarian hyperandrogenism, an endocrinopathy related to reduced fetal growth, but the characteristics of their ovulatory function have not been fully documented. We assessed ovulatory function by weekly urinary LH and salivary progesterone measurements over 3 consecutive months in 85 adolescent girls with known weight and gestational age at birth: 49 girls had no history of PP (age, 14.7+/-1.7 yr), and 36 had a history of PP (age, 14.4+/-2.0 yr); 55 girls were in the early postmenarcheal phase (0-3 yr after menarche), and 30 were in the late postmenarcheal phase (> 3 yr after menarche). In girls with PP, the 17-hydroxyprogesterone (17-OHP) response to ACTH was determined at prepubertal diagnosis of PP, and serum androgen and gonadotropin concentrations were measured in adolescence together with insulin responses to an oral glucose load. Early postmenarche, the fraction of girls with ovulations was similar in the non-PP and PP subgroups (61% vs. 62%), as was the fraction of ovulatory cycles (25% vs. 22%). Late postmenarche, however, the fractions of ovulating girls and ovulatory cycles were strikingly higher (P < or = 0.001) in the non-PP than in the PP subgroup (91% vs. 20% and 47% vs. 12%). Within the PP subgroup, anovulatory girls were found to have a lower weight SD score at birth (mean+/-SEM) than ovulatory girls (-1.22+/-0.3 vs. -0.36+/-0.3; P = 0.03), a higher 17-OHP response to ACTH before puberty (333.1+/-31 vs. 203.8+/-26 ng/dL; P < 0.002), and, in adolescence, lower serum sex hormone-binding globulin levels and higher circulating LH, free androgen indexes, and insulin responses. In conclusion, these findings indicate that girls with PP are at increased risk for anovulation from late (not early) adolescence onward, particularly those girls with a low weight at birth and/or a high 17-OHP response to ACTH at prepubertal diagnosis of PP.

Prospective, randomized, crossover study to evaluate the benefit of human chorionic gonadotropin–timed versus urinary luteinizing hormone–timed intrauterine inseminations in clomiphene citrate–stimulated treatment cycles

Objective: To compare two methods of timing IUI, urinary LH monitoring and transvaginal ultrasonography/hCG timing of ovulation, in patients receiving clomiphene citrate. Design: Prospective, randomized, crossover study. Setting: Yale University Reproductive Medicine Center. Patient(s): Infertile couples undergoing IUI because of unexplained infertility, anovulation, or male factor infertility. Intervention(s): Patients received clomiphene citrate on days 3–7 of the menstrual cycle and were randomized initially to one of two monitoring protocols. In protocol A, urinary LH monitoring was used to time IUI. Urinary LH levels were determined daily with the use of commercial kits, starting on day 10 of the cycle. When urinary LH was detected, IUIs were performed daily for the next 2 days. In protocol B, ultrasound monitoring of folliculogenesis was performed until a leading follicle of ≥18 mm was noted, at which time hCG (10,000 IU) was given intramuscularly and IUIs were performed daily for the next 2 days. If no pregnancy occurred, the couple crossed over to the alternate protocol for the next cycle and continued this alternating therapy for a total of four cycles. Main Outcome Measure(s): Pregnancy rate per cycle. Result(s): One hundred forty-one cycles were completed. In these cycles, six pregnancies occurred, for an overall pregnancy rate of 4.26% per cycle. The pregnancy rate with LH-timed IUI was 4.29% (3/70) and that with hCG-induced ovulation was 4.23% (3/71); the difference was not statistically significant. Conclusion(s): Timing IUI with the use of a relatively expensive and time-consuming method such as ultrasound monitoring of folliculogenesis and hCG induction of ovulation does not appear to produce an increased pregnancy rate over urinary LH monitoring of ovulation.