Characterization of the Antarctic sea urchin (Sterechinus neumayeri) transcriptome and mitogenome: a molecular resource for phylogenetics, ecophysiology and global change biology.

This is the first de novo transcriptome and complete mitochondrial genome of an Antarctic sea urchin species sequenced to date. Sterechinus neumayeri is an Antarctic sea urchin and a model species for ecology, development, physiology and global change biology. To identify transcripts important to ocean acidification (OA) and thermal stress, this transcriptome was created pooling, and 13 larval samples representing developmental stages on day 11 (late gastrula), 19 (early pluteus) and 30 (mid pluteus) maintained at three CO2 levels (421, 652, and 1071 µatm) as well as four additional heat-shocked samples. The normalized cDNA pool was sequenced using emulsion PCR (pyrosequencing) resulting in 1.34M reads with an average read length of 492 base pairs. 40,994 isotigs were identified, averaging 1188 bp with a median coverage of 11×. Additional primer design and gap sequencing were required to complete the mitochondrial genome. The mitogenome of S. neumayeri is a circular DNA molecule with a length of 15 684 bp that contains all 37 genes normally found in metazoans. We detail the main features of the transcriptome and the mitogenome architecture and investigate the phylogenetic relationships of S. neumayeri within Echinoidea. In addition, we provide comparative analyses of S. neumayeri with its closest relative, Strongylocentrotus purpuratus, including a list of potential OA gene targets. The resources described here will support a variety of quantitative (genomic, proteomic, multistress and comparative) studies to interrogate physiological responses to OA and other stressors in this important Antarctic calcifier.

Immunohistochemical localization of epidermal growth factor in human endometrium, decidua, and placenta.

Epidermal growth factor (EGF) was localized immunohistochemically in human endometrium throughout the menstrual cycle, in gestational decidua, and in first, second, and third trimester placenta using two polyclonal antihuman EGF antisera. In proliferative phase endometrium, moderate EGF immunostaining was localized to the cytoplasm of stromal cells, with absent to light staining of glandular epithelium. In the secretory phase, EGF immunostaining was intense and localized predominantly to stromal cells, particularly those surrounding spiral arterioles. There was absent to light EGF immunostaining within epithelial cells; however, there was no staining of subnuclear vacuoles. In addition, the luminal surface of exhausted secretory glands demonstrated moderate EGF immunostaining. In gestational decidua, EGF immunostaining was light to moderate in the stromal cells, but was intense in the surface epithelium. Intense EGF immunostaining was noted in the syncytiotrophoblast layer of first trimester placenta, with light to moderate staining of the cytotrophoblast. Immunostaining decreased in both layers of trophoblast as pregnancy progressed. Immunoreactive EGF is found in endometrium and trophoblast and may have a physiological role in endometrial and placental function.

Transforming growth factor-alpha in human implantation trophoblast: immunohistochemical evidence for autocrine/paracrine function.

Epidermal growth factor (EGF) and its receptor (EGF-R) have been demonstrated in human implantation sites. Transforming growth factor-alpha (TGF-alpha), a protein with extensive sequence homology to EGF and with equal affinity for the EGF-R, was localized immunohistochemically in early intrauterine and ectopic pregnancies. Within the same experiments, TGF-alpha immunostaining was more intense in ectopic than intrauterine pregnancies. In both groups, TGF-alpha immunostaining was moderate to intense in the syncytiotrophoblast (ST), light to moderate in the cytotrophoblast (CT), and moderate to intense in intermediate trophoblast (IT). In ST, TGF-alpha immunostaining localized to the cytoplasm and plasma membranes, including microvilli. No nuclear associated TGF-alpha was noted in ST. In CT, differential TGF-alpha immunostaining was noted between the villous and nonvillous CT. Villous CT demonstrated light to absent cytoplasmic TGF-alpha immunostaining with intense nuclear staining. In contrast, nonvillous CT revealed moderate to intense cytoplasmic staining without demonstrable nuclear staining. These results demonstrate the presence of immunoreactive TGF-alpha in all forms of trophoblast. The known presence of the EGF-R suggests an autocrine/paracrine role for TGF-alpha during human implantation.

Prostaglandin E and F2 alpha receptors in human uterine leiomyomas.

9uman uterine leiomyomas specifically bound less (P less than 0.01) [3H]prostaglandin E1 ([3H]PGE1) and [3H] PGF2 alpha than adjacent normal myometria [leiomyomas: mean [3H]PGE1, 16.4 (range, 11.1-25.2) fmol/mg protein; mean [3H]PGF2 alpha, 4.7 (range, 0.8-12.1) fmol/mg protein; adjacent normal myometria: mean [3H]PGE1, 41.7 (range 27.1-60.7) fmol/mg protein; mean [3H]PGF2 alpha, 7.8 (range, 4.3-16.3) fmol/mg protein]. The lower binding of both [3H]PGs by leiomyomas was due to lower numbers of available high and low affinity sites. Leiomyomas and normal adjacent myometria bound 4-7 times more [3H]PGE1 than [3H]PGF2 alpha, and this appears to be due to high affinity and high numbers of low affinity PGE sites. The smooth muscle content was lower (P less than 0.01) in leiomyomas (mean, 28.0%; range, 9.7-45.5%) than that of adjacent normal myometria (mean, 58.9; range, 51.4-71.2%). In summary, this is the first demonstration of PGE and PGF2 alpha receptors in human uterine leiomyomas. Lower receptor numbers in leiomyomas appear to be due to the lower smooth muscle content of the tissue.

Binding sites for epidermal growth factor in human uterine tissues and leiomyomas.

There is no published data regarding whether human uterine tissues and leiomyomas contain binding sites for epidermal growth factor (EGF). The present study was undertaken with 33 myometria, 13 leiomyomas, and 4 endometria. All myometria (fundus) and leiomyomas and 3 of 4 endometria specifically bound 125I-labeled mouse EGF (myometria: mean, 1.1; range, 0.1-3.9 fmol/mg protein; leiomyomas: mean, 1.1; range, 0.2-2.6 fmol/mg protein; endometria: mean, 1.0; range, 0.0-3.1 fmol/mg protein). [125I]EGF binding to myometrium was ligand specific in that only unlabeled EGF, but not unlabeled insulin, hCG, human PRL, prostaglandin E1, or prostaglandin F2 alpha, competed with [125I]EGF for binding. The apparent dissociation constants and specific binding capacities for myometrium and leiomyoma were: 0.7 nM, 1.9 fmol/mg protein; and 0.1 and 3.7 nM, 0.1 and 4.4 fmol/mg protein, respectively. While smooth muscle content decreased from the fundus to the cervical end of uteri, [125I]EGF binding did not correspondingly decrease (r = 0.5; n = 4). The binding of [125I]EGF to myometrium did not vary with the phase of the menstrual cycle or the patients' diagnosis before hysterectomy (P greater than 0.1). In summary, these results demonstrate that human uteri and leiomyomas contain specific, high affinity binding sites for EGF, but the binding neither exhibited topographical changes nor varied with the phase of the menstrual cycle or benign pathological state of the tissue.

Immunohistochemical localization of transforming growth factor-beta in human implantation sites.

Transforming growth factor-beta (TGF beta), a protein known to antagonize many of the functions of the epidermal growth factor-receptor system, was localized immunohistochemically in unruptured ectopic pregnancies (EP) removed by salpingectomy (n = 8), uterine decidua from EP (n = 4), and decidua and trophoblast from electively terminated first trimester pregnancies (ETP; n = 8). Two rabbit polyclonal antisera that recognize both TGF beta 1 and beta 2 were used. Immunostaining for TGF beta was identified in all three forms of trophoblast, cytotrophoblasts, intermediate trophoblasts, and syncytiotrophoblasts, which were differentiated histologically and immunohistochemically. Moderate cytoplasmic immunostaining was found in villous cytotrophoblasts in both EP and ETP. Nonvillous (anchoring) cytotrophoblasts in these same tissues demonstrated moderate immunostaining adjacent to the villous and light immunostaining distal to the villous. In intermediate trophoblasts, moderate to intense immunostaining was seen in EP and ETP. Syncytiotrophoblasts demonstrated moderate cytoplasmic immunostaining in EP and ETP as well as moderate to intense staining of plasma membranes and microvilli. Nuclear staining was not evident in any form of trophoblast. TGF beta immunostaining was demonstrated in both glands and stroma of decidua from both EP and ETP; however, staining was more intense in decidua from ETP. With the known presence of TGF beta receptors and mRNA in placenta, these results suggest an autocrine/paracrine role for TGF beta regulation of endometrial-trophoblast function during human implantation.

Epidermal growth factor and its receptor in human implantation trophoblast: immunohistochemical evidence for autocrine/paracrine function.

Epidermal growth factor (EGF) and its receptor (EGF-R) were immunohistochemically localized in trophoblast during human implantation from intrauterine and ectopic pregnancies. EGF immunostaining was absent to light in the cytotrophoblast (CT), light to moderate in intermediate trophoblast (IT), and intense in the syncytiotrophoblast (ST). In ST, EGF immunostaining was found mostly in the cytoplasm; however, staining of the plasma membrane was also noted. Immunostaining for the EGF-R was absent to light in the CT and moderate to intense in the IT. Immunostaining for the EGF-R was intense in the ST, with moderate staining in the cytoplasm and intense staining in the plasma membrane. Staining was most intense on the microvilli of the ST. Additionally, EGF-R immunostaining could be demonstrated on nuclear membranes. The increase in the intensity of the immunostaining for both EGF and EGF-R noted in CT, IT, and ST suggests a differentiated expression of this receptor-ligand system in human trophoblast and provides evidence for an autocrine/paracrine role for EGF in trophoblast function. The presence of this receptor-ligand system during early human implantation strongly supports a role for EGF and the EGF-R in embryo-uterine signalling and the implantation process.

Topography of human uterine prostaglandin E and F2 alpha receptors and their profiles during pathological states.

The topography of prostaglandin (PG) E and F2 alpha receptors in uteri of premenopausal women was investigated by dividing uteri into six equal longitudinal strips and further dividing each strip into approximately 1-cm segments. Tissue for determination of smooth muscle content using the Trichrome stain was taken from each section, and the remainder was homogenized for binding studies with 3H-labeled PGs. The [3H] PGE1 binding (mean, 41.5 fmol/mg protein; range, 23.1-58.3) was about 8-fold greater in the fundus than [3H]PGF2 alpha binding (mean, 4.8 fmol/mg protein; range, 1.3-13.0), and this trend was found in most uterine sections. The binding of both 3H-labeled PGs decreased from fundus to cervix, and this decrease was similar to the decrease in smooth muscle content. Scatchard analysis revealed apparent dissociation constants (Kds) of 1.4 and 76 nM and apparent specific binding capacities (Ns) of 25 and 488 fmol/mg protein for [3H]PGE2, and Kd values of 11.5 and 81 nM and Ns values of 19.4 and 58 fmol/mg protein for [3H]PGF2 alpha in the uterine fundus. The Kd values for [3H]PGE2 were similar in other sections of the uterus, but the Ns values were smaller in the lower uterine body and cervical end. While the phase of the menstrual cycle did not influence [3H]PG binding, the diagnosis of abnormal uterine bleeding compared to dysmenorrhea was associated with an increase in [3H]PGE1 binding (P less than 0.05).

Immunohistochemical localization of transforming growth factor-alpha in human endometrium, decidua, and trophoblast.

Transforming growth factor-alpha (TGF-alpha) was localized immunohistochemically in human proliferative and secretory endometrium, decidua, and trophoblast from first, second, and third trimester pregnancies. In proliferative endometrium, TGF-alpha immunostaining was moderate to intense and localized predominantly to stromal cells, whereas glandular staining was absent to light. After ovulation, TGF-alpha staining was light within the stroma, but moderate to intense around spiral arterioles. Moderate to intense staining was also detected in glandular and surface epithelium in secretory endometrium, with no staining noted in subnuclear vacuoles. In hypersecretory endometrium, staining was predominantly epithelial. In decidua, TGF-alpha was detected in intermediate trophoblast and on the surface epithelium. In first trimester trophoblast, TGF-alpha was detected in both cytotrophoblast (CT) and syncytiotrophoblast. Cytoplasmic staining was light in CT and moderate to intense in ST, with particular staining of plasma membranes. Intense TGF-alpha staining of nuclear membranes in CT was noted. TGF-alpha staining was light to absent in second and absent in third trimester trophoblast. This study demonstrates immunoreactive TGF-alpha in tissues known to be responsive to epidermal growth factor, and also demonstrates the presence of immunoreactive TGF-alpha associated with nuclear membranes. Thus, TGF-alpha may play an autocrine/paracrine role in endometrial development and trophoblast function.

An insight into early reproductive processes through the in vivo model of ovum donation.

To gain insight into early reproductive processes we have prospectively designed ovum donation protocols to elucidate several phenomena relating to embryo implantation and pregnancy sustenance. Artificial endometrial cycles with variable follicular phases were induced in 60 recipients by sequential estrogen and progesterone. A total of 964 oocytes were retrieved throughout 43 ovum donation attempts, for an average of 22.4 (range, 16-41) eggs/retrieval. The overall delivery rate per egg retrieval (donors and recipients combined) was 72.1% (31 of 43). The shortest estrogen stimulation (short follicular phase) resulting in ongoing pregnancies was 5 days in duration, while the longest (long follicular phase) was 35 days in duration before progesterone initiation. Utilization of variable length follicular phases, artificially extended the stage of endometrial receptivity to over 4 weeks. To assess the window of implantation, same age embryos were transferred onto endometrium of different maturational stages. Pregnancies were documented with embryo transfers between luteal day 1 (day 15) to luteal day 6 (day 20), extending the window of implantation in the human to at least 6 consecutive days. To evaluate the relative contribution of oocyte quality and endometrial receptivity to pregnancy outcome, common source ova were transferred onto endometrium with variable hormonal exposure. Despite the drastically different follicular phase estradiol levels and periods of exposure, similar delivery rates were attained in donor cycles (29.4%) and recipient cycles during short follicular phases (29.6%). Slightly higher delivery rates (39.4%) were observed with long follicular phases. The comparable pregnancy rates in donors and recipients are attributed to the common source oocytes regardless of endometrial stimulation.

Epidermal growth factor in urine of nonpregnant women and pregnant women throughout pregnancy and at delivery.

Human epidermal growth factor (EGF) concentrations were measured by a specific solid phase RIA in random urine samples collected throughout the menstrual cycle of normal menstruating women (n = 8), women with tubal sterilization (n = 6), women taking a low dose oral contraceptive (n = 5), and women throughout pregnancy (n = 52) and delivery (n = 35). There were no differences in EGF concentrations between the proliferative and secretory phases of the menstrual cycle (P greater than 0.05). Normal menstruating women had higher urinary EGF concentrations [mean +/- SE, 37.2 +/- 6.0 micrograms/g creatinine (4.23 +/- 0.68 ng/mumol)] than women with tubal sterilization [32.7 +/- 4.0 (3.71 +/- 0.45)] or women taking a low dose oral contraceptive [19.5 +/- 6.0 (2.21 +/- 0.68)], but the differences were not significant (P greater than 0.05). During pregnancy, urinary EGF concentrations increased linearly from 6-20 weeks gestation (r = 0.76; P less than 0.001), then declined toward term (r = -0.71; P less than 0.001). EGF concentrations in early pregnancy (less than 12 weeks) or at term did not differ significantly from those in normal menstruating women (P greater than 0.05). For women delivering normal, appropriate for gestational age (AGA) infants, there was no correlation between urinary EGF concentrations and fetal weight or sex (P greater than 0.05). Urinary EGF concentrations in women delivering normal AGA infants [52.7 +/- 2.5 (5.98 +/- 0.28); n = 16] did not differ significantly (P greater than 0.05) from those in women with class A/B diabetes [41.9 +/- 2.8 (4.76 +/- 0.31); n = 6] or women delivering twins [45.6 +/- 2.6 (5.18 +/- 0.29); n = 8] with a greater fetoplacental mass. However, women delivering an intrauterine growth-retarded fetus with decreased fetoplacental mass had lower urinary EGF concentrations (24.9 +/- 2.2 (2.83 +/- 0.25); n = 5] than women with normal AGA infants (P less than 0.01). The significance of the rise in the urinary EGF concentration late in the second trimester and lower urinary EGF concentrations in women delivering intrauterine growth-retarded infants is not known, but may reflect an important physiological role for EGF in fetal-maternal hormonal interaction and development.

EPR investigation of the Mn(II) binding sites in glutamine synthetase (Escherichia coli W). II. Intermediate-affinity binding sites.

The nature of the intermediate-affinity (n2) Mn(II) binding sites in glutamine synthetase [EC 6.3.1.2] has been studied as a function of adenylylation in a variety of enzyme-metal complexes by EPR. In the absence of nucleotide the n2 Mn(II) environment is nearly isotropic, the Mn(II) bonds are highly ionic, and the interaction distance R greater than or equal to 12-14 A. Nucleotide binding at the n2 Mn(II) site renders the n2 Mn(II) signal unobservable and causes a reduction in signal amplitude (approximately 30%) and line broadening (approximately 6 G) at the high-affinity (n1) Mn(II) site. This behavior indicates that nucleotide binding induces a conformational change in the enzyme which brings the previously distant n1 and n2 sites into closer proximity (R less than or equal to 8-11 A), possibly for the purpose of activating the nucleotide for direct phosphoryl transfer to L-glutamate. In line with this suggestion, the broad, unresolved resonances in complexes containing both L-methionine SR-sulfoximine (MSOX) and nucleotide may result from the phosphorylation of MSOX. The n2 Mn(II) site is not affected by adenylylation in all the enzyme-metal complexes studied, which suggests that the regulatory effects of adenylylation may only act at the n1 Mn(II) sites.

Hysterosalpingographic diagnosis of uterine adenomyoma.

True adenomyomas (encapsulated) are uncommon tumors of the uterus. At hysterosalpingography, detection of a network of fine channels in a very well-circumscribed area of the myometrium, connected with the uterine cavity, allows a preoperative diagnosis.

A prospective evaluation of clomiphene citrate challenge test screening of the general infertility population.

OBJECTIVE: To assess the results of screening an unselected general infertility population for diminished ovarian reserve with the clomiphene citrate challenge test. METHODS: Two hundred thirty-six couples were followed prospectively and studied for the relationship between clomiphene citrate challenge test screening and final diagnoses and long-term fertility rates. RESULTS: Abnormal clomiphene citrate challenge tests were found in two of 61 (3%) of the patients younger than 30 years, in five of 72 (7%) aged 30-34, in seven of 68 (10%) aged 35-39, and in nine of 35 (26%) aged 40 or older. An abnormal test predicted lower pregnancy rates; conception occurred in 92 of 213 (43%) of patients with normal results, but only two of 23 (9%) of patients with abnormal results (P < .004). Unexplained infertility (not considering the clomiphene citrate challenge test) was a common finding in patients with abnormal clomiphene citrate challenge test results (12 of 23). This incidence was significantly higher than that in patients with normal clomiphene citrate challenge test results (20 of 213) (P < .001). CONCLUSIONS: Approximately 10% of the patients in the general infertility population had abnormal clomiphene citrate challenge tests. The incidence of abnormal results increases with age beginning in the early 30s, occurs with a higher frequency in patients who would otherwise be diagnosed with unexplained infertility, and prognosticates decreased long-term pregnancy rates.

Prognostic assessment of ovarian reserve.

OBJECTIVE: To review the literature regarding diminished ovarian reserve, the screening techniques that are currently available, and their appropriate application in clinical practice. DATA RESOURCES: Directed Medline searches. RESULTS: Ovarian reserve screening identifies women with greatly diminished chances of achieving pregnancy. The screening techniques include the clomiphene citrate challenge test, basal day 3 FSH measurements, and the GnRH agonist stimulation test. All have been evaluated in assisted reproduction programs and the predictive values of an abnormal test for failing to conceive is very high. When abnormal, these tests allow physicians to counsel patients that their prognosis for conception is poor. Although the presence of a normal result does indicate better long-term chances for conception, an age-related decline in fecundity remains and patient age should still be considered when counseling patients with normal screening results. Clinicians are urged to validate the threshold values with the assay system used in their own laboratory before the application of these tests. CONCLUSION: The literature consistently demonstrates the value of diminished ovarian reserve screening.

Recurrent pregnancy loss and diminished ovarian reserve.

OBJECTIVE: To determine the incidence of diminished ovarian reserve (OR) in patients with recurrent pregnancy loss (RPL). DESIGN: Retrospective chart review. SETTING: Tertiary fertility center. PATIENT(S): Six hundred ninety-two women undergoing a fertility evaluation. INTERVENTION(S): Clomiphene citrate challenge test (CCCT). MAIN OUTCOME MEASURE(S): FSH concentrations measured on menstrual days 3 and 10. RESULT(S): Forty-four women were diagnosed with RPL (+RPL), and 648 women had non-RPL diagnoses (-RPL). Compared with -RPL women, women with +RPL were younger (following statistics are listed as +RPL vs. -RPL, respectively; 34 +/- 5 vs. 35 +/- 4 y) but had similar menstrual cycle length (29 +/- 4 vs. 28 +/- 4 d), and lower day 3 FSH levels (8.9 + 7 vs. 11 +/- 9 mIU/mL) and similar day 10 FSH levels (11 +/- 8 vs. 12 +/- 11 mIU/mL). Eight of 44 women with +RPL (18%) had an abnormal CCCT, compared with 117/648 (18%) of women in the -RPL group. For women with normal OR, delivery rates were similar for -RPL and +RPL patients. For women with an abnormal CCCT, delivery rates were < 5%. CONCLUSION(S): Women with RPL have a similar incidence of diminished OR as the general infertile population. Reproductive outcome for patients with an abnormal CCCT is equally poor for both groups. Ovarian reserve screening should be considered in the work-up of RPL before initiation of anticoagulant or immunotherapy.

Intercycle variability of day 3 follicle-stimulating hormone levels and its effect on stimulation quality in in vitro fertilization.

Prior studies have demonstrated that gonadotropin stimulation quality and pregnancy rates are better in in vitro fertilization (IVF) patients with low basal cycle day 3 follicle-stimulating hormone (FSH) levels. The records of 81 patients who had undergone three or more IVF attempts during a 2-year period were studied to determine the degree and potential impact of intercycle variability in basal FSH concentrations. The mean of the individual standard deviations for all 81 patients was 4.2 +/- 0.4 mIU/mL. However, the patients with a mean basal FSH of less than 15 mIU/mL had a mean deviation of only 2.6 +/- 0.2 mIU/mL, whereas those with a mean basal FSH of greater than or equal to 15 mIU/mL had a mean deviation of 7.3 +/- 0.7 mIU/mL. Intercycle variability in basal FSH values did not predict changes in ovarian response to gonadotropin stimulation and thus may not be used to select an optimal cycle in which to stimulate an individual patient. Furthermore, patients with large intercycle variation responded poorly to gonadotropin stimulation independent of their basal FSH concentration. This information allows more precise counseling of patients regarding their appropriateness for assisted reproduction.

Inhibin-B: the physiologic basis of the clomiphene citrate challenge test for ovarian reserve screening.

OBJECTIVE: To determine inhibin-B concentrations during ovarian reserve screening in women with normal and diminished ovarian reserve as determined by the clomiphene citrate challenge test. DESIGN: Retrospective. SETTING: Tertiary fertility center. PATIENT(S): Women undergoing ovarian reserve screening for a routine fertility evaluation. INTERVENTION(S): Clomiphene citrate challenge test. MAIN OUTCOME MEASURE(S): Inhibin-B concentrations on menstrual days 3 and 10. RESULT(S): Nineteen patients with normal ovarian reserve and 15 with diminished ovarian reserve had serum inhibin-B concentrations determined during ovarian reserve screening. For all patients, day 10 inhibin-B concentrations were higher than day 3. Women with normal ovarian reserve had higher inhibin-B concentrations on both days 3 and 10 than women with diminished ovarian reserve. Inhibin-B concentrations demonstrated a negative correlation with FSH levels on both cycle days 3 and 10 and a positive correlation with E2 on cycle day 10. CONCLUSION(S): Women with diminished ovarian reserve during ovarian reserve screening had reduced granulosa cell inhibin-B production compared with women with normal ovarian reserve. The lower inhibin-B concentrations may be responsible for the elevated FSH concentrations and may be indicative of the aging follicular apparatus.

Endometrial thickness is predictive of histologic endometrial maturation in women undergoing hormone replacement for ovum donation.

OBJECTIVE: To determine if ultrasonographic endometrial pattern or thickness is predictive of histologic endometrial maturation in women undergoing hormone replacement for ovum donation. DESIGN: Ultrasonographic endometrial thickness and pattern were determined and compared with histologic assessment of endometrial maturation. PATIENTS: Forty-six women underwent 52 preparatory cycles for ovum donation. Transvaginal ultrasound (US) was performed after 14 days of E2 replacement and, after 12 days of P, an endometrial biopsy was performed. In 12 cycles, a continuous dose of 2 mg/d E2 was administered. In cycles with out-of-phase biopsies (dated earlier than day 24) and in the last 34 cycles, all women received an escalating dose of E2 before initiation of P. Additionally, the 46 women underwent 55 ETs with USs performed on cycle day 15. RESULTS: Six women had abnormal biopsies in their first preparatory cycle on the continuous E2 protocol, which normalized with the escalating protocol. All other women had normal biopsies. Women with abnormal biopsies had significantly thinner endometrium (< or = 6 mm) but similar endometrial patterns compared with women with normal biopsies. In women having US in preparatory and transfer cycles, there were no differences in endometrial thickness or pattern between examinations. CONCLUSIONS: Endometrial thickness > or = 7 mm in hormone replacement cycles predicts in phase endometrial histology and can replace the endometrial biopsy.

Efficacy of selection criteria for ovarian reserve screening using the clomiphene citrate challenge test in a tertiary fertility center population.

OBJECTIVE: To examine selection criteria for ovarian reserve screening. DESIGN: Retrospective study. PATIENTS: Two hundred nineteen women underwent testing for ovarian reserve for woman's age > 35 years, any age with unexplained infertility, one ovary, or a poor response to hMG. INTERVENTIONS: Clomiphene citrate challenge test. MAIN OUTCOME MEASURES: Frequency of abnormal ovarian reserve screening, menstrual cycle parameters, response to hMG, and pregnancy outcome by screening criteria. RESULTS: One hundred eighty-four (84.0%) women had a normal ovarian reserve screening test; 35 (16.0%) had an abnormal ovarian reserve screening test. Twenty-six had abnormal ovarian reserve screening when screened by age, 14 for unexplained infertility, 5 for poor response to hMG, and 6 for one ovary. Fifteen women with abnormal ovarian reserve screening had more than one indication for screening. For women attempting pregnancy (n = 182), 49 of 148 (33.1%) with normal ovarian reserve screening became pregnant compared with 2 of 34 (5.9%) with abnormal ovarian reserve screening. Within each screening category, women with abnormal ovarian reserve had menstrual cycle parameters associated with a short follicular phase, required more hMG, and responded poorly to hMG. CONCLUSIONS: One of six women undergoing ovarian reserve screening had an abnormal test, which was associated a poor reproductive outcome. Age was the most important single criteria. Selected ovarian reserve screening is simple and inexpensive and should be offered to all fertility patients meeting the specific screening criteria listed above.