Days Of E2 Research Articles

Effect of Short‐Term Estradiol Exposure on Endothelin‐B Receptor‐Mediated Vasodilation in Postmenopausal Women

Endothelin (ET)‐1 contributes to endothelial dysfunction in postmenopausal women (PMW) via the ETB receptor. We have previously shown that the ETB receptor mediates vasodilation in young women, an effect that is lost in PMW. However, it is unclear if ETB receptor‐mediated dilation can be restored in PMW exposed to E2.PURPOSETo test the hypothesis that E2 exposure restores ETB receptor‐mediated vasodilation in PMW.METHODSWe tested 9 PMW (55±2 yrs, 24±3 kg/m2, 91±5 mmHg) who self‐reported natural menopause and were at least 1 year without menses. We measured vasodilation in the cutaneous microcirculation via local heating using laser Doppler flowmetry coupled with intradermal microdialysis during perfusions of lactated Ringer’s (control), ETB receptor antagonist (BQ‐788, 300nM) and ETA receptor antagonist (BQ‐123, 500nM). Cutaneous vascular conductance (CVC) was calculated during the plateau phase of local heating (42°C), and normalized to maximal vasodilation achieved by perfusion of sodium nitroprusside (28mM) and heating to 43°C. All women were tested at baseline (BL) and after 7 days of E2 (Vivelle‐Dot, 0.1mg/day, transdermal patch) administration (+E2). A two‐way repeated measures ANOVA was performed to compare %CVCmax in PMW before and after E2 exposure. Data presented are mean ± standard deviation.RESULTSE2 administration increased serum estradiol (BL: 45±29 vs +E2: 71±42 pg/mL, P<0.01) and reduced follicle stimulating hormone (BL: 71±13 vs +E2: 57±17 pg/mL, P<0.01), but there were no changes in serum progesterone (BL: 0.57±0.93 vs +E2: 0.53±0.88 pg/mL, P=0.14) nor plasma ET‐1 (BL: 1.62±0.66 vs +E2: 1.54±0.54 pg/mL, P=0.42). E2 administration had no effect on cutaneous microvascular responses (Time P=0.41, Drug P=0.17, Interaction P=0.71). At BL, vasodilatory responses to local heating were not different during control and ETB receptor antagonism (control: 83±8 vs. BQ‐788: 88±6 %CVCmax). Similarly, vasodilation was not altered at BL during ETA receptor antagonism in a subset of women (n=6, control: 83±8 vs. BQ‐123: 87±8 %CVCmax). E2 administration did not improve vasodilation in the control site (BL: 83±8 vs. +E2 85±13 %CVCmax). Furthermore, there was no effect of E2 exposure on cutaneous vasodilatory responses during perfusion of ETB (BL: 88±6 vs. +E2: 89±6 %CVCmax) nor ETA (n=6, BL: 87±8 vs +E2: 92±7 %CVCmax) receptor antagonists.CONCLUSIONThese preliminary data suggest that short‐term E2 exposure does not modulate ETB receptor function in PMW. These findings are in contrast to our observations in young women demonstrating that ETB receptors mediate vasodilation in the presence of E2. The role of alternative therapies to modulate ETB receptor function should be considered in future studies.

TO EVALUATE IVF OUTCOME BETWEEN FLARE-UP VERSUS ANTAGONIST AMONG POOR RESPONDERS’ PROGNOSIS PATIENTS AND RELATED FACTORS AT NATIONAL CENTER FOR ASSISTED REPRODUCTIVE TECHNOLOGY

Objectives: To assess the outcome of Flare-up versus Antagonist protocol and to determine factors related to poor responders.Methodology: this is a random control trial among 834 patients who were predicted to ovarian poor response from 2014 to 2018.Results: The rate of biochemical pregnancy was 4.5% with Flare-up versus 8,1% with Antagonist (p<0.05). The rate of fertilization, implantation and clinical pregnancy were not significant differentbetween the 2 protocols. Age and number of AFC were significant factors to predict poor ovarian response. However, with Flare-up, there were 2 more factors could be used for the purpose whichwas basal FSH and E2 day 7.Conclusion: The outcome of treatment between Flare-up and Antagonist among predicted poor ovarian response was comparable. Age and AFC were valuable factors in prediction of poor ovarianresponse.

Assessment of Estradiol-Induced Gonadotropin Surge Generation: Preliminary Results From a Protocol of Graded Transdermal Estradiol Dosing

In normally-cycling women, high midcycle estradiol (E2) concentrations (> 200-300 pg/ml) derived from the dominant follicle trigger an approximately 10-fold increase in LH and 4-fold increase in FSH levels—the gonadotropin surge—over about 48 hours. In prenatally-androgenized female rats and sheep (animal models with polycystic ovary syndrome [PCOS]-like features), high-dose estrogen fails to initiate gonadotropin surges. However, it remains unclear whether E2-induced gonadotropin surge generation is defective in PCOS. Baird et al., induced LH surges in 12 women with PCOS and 6 normally-cycling controls (studied in the early follicular phase) with 3 days of high-dose oral ethinyl E2, and the LH peak magnitude was similar in both groups (JCEM 1977;45:798-801). However, the dose of ethinyl E2 in this study (200 mcg/day) was markedly supraphysiological, and the dosing schedule did not mimic the normal, gradual increase in E2 levels observed in normal follicular phases. To further assess potential impairments in E2 augmentation of gonadotropin secretion in PCOS, we developed a study protocol involving graded transdermal E2 dosing (with dose adjustments as needed) to gradually achieve serum E2 concentrations approximating 300-400 pg/ml. Herein we present our early experience with this protocol in six normally-cycling, non-hyperandrogenic women: median (range) age 23.1 (19.8–29.0) years, body mass index 20.7 (19.2–27.2) kg/m2, calculated serum free testosterone 2.7 (2.4–5.3) pg/ml. Study subjects began transdermal E2 on cycle day 4 with the following dose-escalation protocol: 0.2 mg/d x 1 day, then 0.3 mg/d x 1 day, then 0.4 mg/d x 1 day, then 0.5 mg/d. Daily serum E2 measurements informed dose adjustments (e.g., a measured E2 level > 400 pg/ml prompted a dose reduction; a measured E2 level < 250 pg/ml while on the 0.5 mg/d dose prompted a dose increase to 0.6 mg/d). In these six women, daily E2 values (pg/ml) were as follows: baseline 31 (17–36) (median [IQR]) on cycle day 4 (the morning of E2 initiation); 205 (135–240) on day 5; 159 (135–270) on day 6; 263 (202–363) on day 7; 276 (239–351) on day 8; 328 (307–367) on day 9; 333 (269–402) on day 10; and 328 (253–354) on day 11. Median transdermal E2 doses (mg/d) on days 5 through 11 were 0.2, 0.3, 0.4, 0.35, 0.4, 0.4, and 0.4, respectively. Morning serum LH concentrations (median [IQR]) were 5.2 (3.5–6.1) IU/L immediately before E2 initiation (cycle day 4), decreased to a nadir of 1.5 (1.0–2.4) IU/L after 2 days of E2 (cycle day 6), thereafter increasing 8.1-fold to a peak of 12.1 (8.1–18.3) IU/L after 5 days of E2 (cycle day 9). FSH similarly changed from baseline median 3.5 (3.4–4.2) IU/L to nadir median of 1.6 (1.2–1.8) IU/L after 2 days of E2, thereafter increasing again to 3.5 (2.6–4.0) IU/L after 5 days of E2. We conclude that this experimental protocol may be useful to investigate potential defects in E2-induced LH surge generation in PCOS.

Prediction of severe ovarian hyperstimulation syndrome in women undergoing in vitro fertilization using day 3 estradiol levels, collected ova, and the number of follicles

Ovarian hyperstimulation syndrome (OHSS) is a potentially life-threatening iatrogenic condition that can occur during in vitro fertilization (IVF). The worst outcome is hydrothorax, hypovolemia, higher risk of deep venous thrombosis and oliguria. With severe OHSS patients are required to postpone embryo transfer for an undetermined amount of time. Predicting which patients are a high risk of developing OHSS is mainly based on serum estradiol (E2) levels, but other factors, such as female age, BMI, ovarian volume, antral follicle count (AFC), and polycystic ovary syndrome, are speculated to be predictive. Here, we aimed to determine if E2 levels at Day 3 and its fold change at day ten as well as antral follicle count and ova collected are predictive factors for severe OHSS. Retrospective case-control study. Patient chart review was performed between January 2008 and December 2017 at Ingenes in Mexico City. Three hundred twenty-seven women were selected. E2 >3000 ng/L usually on the last day of stimulation (day 10 with three 18 mm-follicles) was defined as OHSS (n=151). Culdocentesis was performed on a patient when upon clinical assessment patient presented features such as nausea, vomiting, oral intolerance and ascites identified by endovaginal ultrasound and abdominal ultrasound (renal and hepatic areas with visible ascites) that do not respond to conservative management (n=55 severe OHSS). Predictability was evaluated by measuring the area under the receiver-operating characteristic (AUC). Differences between groups were determined by t-test. The OHSS positive group, when compared to the non-OHSS group, was higher with respect to E2 Day 3 levels (150±230 v 250±177 ng/L), E2 fold change (20.1±23.8 v 32.2±29.1), AFC (11.6±8.3 v 18.2±9.1), and Ova collected (10.1±6.4, 21.1±9.0, p<0.001). E2 Day 3 levels (AUC=0.76, 95%CI: 0.71-0.82), E2 fold change (AUC=0.71, 95%CI: 0.65-0.77), AFC (AUC=0.75, 95%CI: 0.70-0.81), and Ova collected (AUC=0.85, 95%CI: 0.81-0.89) were predictive of OHSS. For Culdocentesis, E2 Day 3 levels (190±221 v 232±158 ng/L) were not different between the subjects who received culdocentesis, whereas the E2 fold change (24.5±26.6 v 32.9±28.8, p=0.038), AFC (13.7±9.0 v 19.8±8.9, p<0.001), and Ova collected (13.7±8.9, 23.3±8.1, p<0.001) were higher. Interestingly, all variables were predictive of subjects who would qualify for culdocentesis (E2 Day 3 levels: AUC=0.63, 95%CI: 0.55-0.70; E2 fold change: AUC=0.63, 95%CI: 0.55-0.71; AFC: AUC=0.74, 95%CI: 0.68-0.80; and Ova collected: AUC=0.80, 95%CI: 0.75-0.85). Here, we demonstrate the E2 levels, as well as the ova production parameters, are indicators of IVF patients who could develop severe OHSS and may require culdocentesis.

Effect of randomized serum progesterone concentration on secretory endometrial histologic development and gene expression.

What doses of secretory phase progesterone (P) in women are associated with altered endometrial structure and/or function? Consistently delayed histological maturation was seen at the lowest tested daily P dose (2.5 mg), whereas consistently altered functional response, as reflected by microarray analysis of gene expression was seen at both the 5 and 2.5 mg doses. Progesterone is absolutely required for normal embryo implantation and pregnancy survival. Progesterone supplementation is beneficial in ART cycles. In this case-control experimental trial, 46 healthy young female volunteers (age 19-34) underwent a single modeled endometrial cycle after GnRH down-regulation or monitored in natural cycles. In a university hospital, modeled cycles were obtained by GnRH agonist down-regulation, transdermal estradiol (E2) (0.2 mg/d), and daily injections of P in oil for 10 days: 2.5 mg (n = 6), 5 mg (n = 6), 10 mg (n = 12) or 40 mg (n = 12), after the 10th day of E2. Ten healthy, ovulatory women were used as controls. Endometrial biopsies were obtained on the 10th day of P exposure, or urinary LH surge (in controls). Analysis included histological dating, serum progesterone levels, microarray analysis of the whole genome, RT-PCR, western blot and comparison with the GEO database. In endometrial biopsies, a morphological delay appears in the 2.5 mg/day of P group. Higher sub-physiological levels of P (≥5 mg/day) resulted in normal histology, but aberrant gene expression. P levels required for consistent histological delay were lower than those in all ovulatory women. Gene expression abnormalities occurred at higher sub-physiological P concentrations, without a change in histology, a functional-morphological disassociation. The expression of some endometrial receptivity-associated genes appeared multiphasic, with peak or nadir of mean or median expression levels between the lowest and highest doses, suggesting sustained supraphysiological doses seen in ART treatment cycles may not be optimal. GEO DataSets ID: 200056980; GSE 56980. These results were obtained in fertile women, who may respond differently from infertile subjects. The dose of P required for normal endometrial structure (5 mg/day) corresponds to a P concentration well below that seen in ovulatory women, suggesting that persistently delayed mid-secretory histology cannot be solely due to inadequate P concentrations in an ovulatory cycle. Endometrial gene expression is differentially regulated by different doses of progesterone. The apparent multiphasic response of some genes to P dose suggests the possibility that P concentration kinetics may play a role in normal endometrial preparation for receptivity. These findings strongly confirm that histologic development is not a reliable measure of endometrial P action. Supported by The Eunice Kennedy Shriver National Institute for Child Health and Disease, National Institute of Health, USA (NICHD/NIH) (R01HD067721 and U54HD30476; SLY and BAL) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) 240239/2012-1 (RFS). All authors have no competing interests.

Requirement of heart and neural crest derivatives–expressed transcript 2 during decidualization of human endometrial stromal cells in vitro

To investigate the role of heart and neural crest derivatives-expressed transcript 2 (HAND2) during decidualization of human endometrial stromal cells (ESCs). In vitro experiment. Research laboratory. Twenty-six patients undergoing hysterectomy for benign reasons. ESCs were cultured for 12 days with HAND2 small interfering RNA (siRNA) or nonsilencing RNA during decidualization by medroxyprogesterone acetate (MPA) and E2. Decidualization was monitored by changes in cellular morphology and the expression of several decidual-specific genes. HAND2 siRNA effectively suppressed HAND2 levels in ESCs after 12 days of E2 + MPA treatment. ESCs cultured with HAND2 siRNA retained a long fibroblast-like shape, whereas the cells cultured with control siRNA transformed into enlarged polygonal cells. Silencing of HAND2 expression significantly reduced connexin-43 involved in the morphologic changes. HAND2 silencing significantly reduced the mRNA levels of fibulin-1, prolactin, tissue inhibitor of metalloproteinase 3, interleukin-15, and forkhead box O1A (FOXO1A), but had no effect on the mRNA levels of dickkopf-1, serum glucocorticoid kinase 1, and insulin-like growth factor-binding protein 5. HAND2 siRNA effectively suppressed the levels of nuclear FOXO1A protein as a regulator of decidualization. These results suggest that HAND2 plays a key role in the regulation of progestin-induced decidualization of human ESCs.

Motherhood after Malignancy: Embracing Oocyte Banking (OB) as a Means to Preserve Fertility between Diagnosis and Cure

and outcomes Age (y) (mean) E2 Day of ov trigger (pg/ml) Total no. retrieved eggs Total no. MII eggs had such a prediction based on age alone. Compared with age-based controls, PredictIVF-D showed 75% improved predictive power, ability to discriminate cases with differential prognoses (10% improvement in area-underthe-curve in a receiver operating characteristic analysis), while extending the dynamic range from four discrete, age-based probabilities to a continuous spectrum ranging from 7 to 60%. Table 1 shows the population distribution and error for five different prognostics groups. CONCLUSION(S): Using a validated, multi-center IVF prediction model, we showed that a large portion of patients with prior IVF would receive a significantly different predicted probability compared with agebased counseling, at improved predictive power and discrimination, and reliable accuracy.

Prophylactic cerclage in twin pregnancies from art: obstetric outcomes

To compare the obstetric outcomes of twin pregnancies after ART with and without prophylactic cerclage (McDonald technique). Cohort prospective study. A total of 129 patients were included, all of them carrying a twin pregnancy resulted from ART between years 2007-2010. Prophylactic cerclage was performed in 46 while the remaining 83 served as controls. The decision about performing or not the cerclage was taken randomly by each attending physicians and all the patients were followed up to delivery. All the demographic characteristics between groups (cerclage and no cerclage) were comparable: age (33.1 ± 5.4 vs 33.8 ± 5.21), weight (68.1 ± 13.7 vs 67.5 ± 12.32kg), height (1.59 ± .066 vs 1.56 ± .077mts), BMI (26.4 ± 5.00 vs 23.0 ± 1.94), FSH day 3 (7.6 ± 6.04 vs 10.49 ± 12.2 IU), LH day 3 (4.03 ± 2.94 vs 5.81 ± 5.23 IU), E2 day 3 (57.6 ± 47.9 vs 57.07 ± 60.6 pg/dL) and PRL (17.46 ± 7.60 vs 16.96 ± 12.32 ng/ml). Similarly there were no differences in day of retrieval (12.67 ± .93vs12.59 ± 6.11), aspirated oocytes (12.83 ± 5.3vs11.70 ± 7.23), MII oocytes (10.32 ± 5.30vs9.77 ± 6.73), injected/inseminated (10.86 ± 5.43vs9.65 ± 5.79), fertilization rate (64%vs74%), day of transfer (2.96 ± 0.81vs2.95 ± 0.72), embryos transferred (2.87 ± 0.65vs3.02 ± 0.62). Among the obstetric variables, a significant difference was observed in gestational weeks (35.65 ± 1.96 vs. 33.79 ± 5.28,P<0.05), average weight (2358.8 ± 462.73 vs.2103.90 ± 711.78, P<0.05). Patients with twin pregnancy and prophylactic cerclage had a better gestational age and better birth weight compared to those without cerclage. Prophylactic cervical cerclage should be considered as routine in twin pregnancies from ART.

Profound pituitary suppression following oral contraceptive pretreatment in GnRH antagonist cycles does not impact outcome

To evaluate the effect of profound pituitary suppression after OCP pretreatment in women undergoing IVF using gonadotropin releasing hormone (GnRH) antagonists. Retrospective cohort study in an academic IVF center. 318 women underwent an IVF cycle utilizing OCP pretreatment with GnRH antagonist protocol from 1/2008 to 1/2010. Patients were assigned to groups based on endogenous LH level on day 1 of stimulation (4 days after OCP discontinuation). Chi-square, Students t-tests, and Pearson's correlation test were used for statistical analysis, p<0.05 considered significant. 75 women had LH levels < 1.5 mIU/mL and 243 women had LH levels > 1.5 mIU/mL. Fertilization, implantation, pregnancy, and spontaneous loss rate were similar in both groups. Significantly greater numbers of oocytes were retrieved in the LH <1.5 mIU/mL group. Longer OCP treatment was correlated with lower LH levels on first day of gonadotropin stimulation (r = −0.175, P=0.002). Greater LH suppression was correlated with increased total gonadotropin dose (r = −0.227, P<0.0001) and days of stimulation (r = −0.445, P<0.0001).Tabled 1LH < 1.5mIU/mLLH > 1.5mIU/mLP valueAge35.9 ± 4.236.5 ± 4.50.31No. days on OCPs22.4 ± 4.521.3 ± 4.20.05Cancellation rate (%)16 (12/75)12.4 (31/243)0.37Total gonadotropin use (IU)4328 ± 12663543 ± 1283<0.0001No. days of stimulation10.9 ± 0.89.5 ± 1.1<0.0001Peak E2 day of hCG (pg/mL)2091 ± 8272049 ± 9900.75Oocytes retrieved (n)17.7 ± 9.414.9 ± 7.80.02Fertilization rate/oocyte(%)56.554.90.38Implantation rate/embryo (%)26.825.60.33Clinical PR/ET(%)49.246.20.79SAB rate (%)19.312.20.48 Open table in a new tab Women with profound LH suppression following OCP pretreatment demonstrate a comparable prognosis to women who did not, despite requiring longer stimulations and a higher total gonadotropin dose. Longer OCP pretreatment results in greater LH suppression. It is possible that OCP pretreatment contributed to synchronization of the early follicular cohort leading to a greater number of oocytes retrieved.

Assessment of Daily Menstrual Blood Loss in Rhesus Macaques.

Rhesus macaques exhibit 28-day menstrual cycles similar to those of women. Menstruation can be experimentally induced by progesterone (P) withdrawal in estradiol (E2) plus P primed macaques and women. However, little is known regarding daily menstrual blood loss (MBL) in hormone controlled subjects. To fill this gap, we tested the effect of artificially short and extended menstrual cycles on daily MBL in macaques. Our goal was to establish baseline daily MBL in macaques for future studies on novel therapies to suppress menses. Twelve ovariectomized rhesus macaques were used in this study. Animal protocols were reviewed and approved by the ONPRC Institutional Animal Care and Use Committee. The animals were treated with an E2-filled Silastic capsule that produced 100-130 pg E2 /ml in serum. After 14 days of E2 priming the animals received a P-filled capsule that produced 5-7 ng P/ml in serum. Removing the P capsule (leaving the E2 capsule in place) on day 28 induced menses (“normal length” cycles). Short 21-day cycles were created by treating animals with E2 for 14 days followed by E2 + P for 7 days. Extended 35 day cycles were induced by treating animals for 14 days with E2, followed by E2 plus one P implant for 7 days and then E2 plus two P implants for 14 days (producing 10-14 ng P/ml). Daily MBL was quantified by a modified alkaline haematin assay. Menstrual blood was collected daily by tampon. On light menstrual flow days Tampex Pearl "Lite" tampons (1.2 cm diameter), cut transversely at 2.5 cm in length, were placed in the vagina to collect menstrual blood. On heavy menstrual flow days Tampex Pearl "Regular" tampons, 3 cm in length, were used. The cut end of the tampons was sealed with Dow Corning Type A Silastic Medical Adhesive and the original tampon applicators were used to insert the tampons into the animals. The tampons were replaced twice each day on normal flow days and 3 times each day on heavy flow days. After collection the tampons were vacuum dried and mixed with 5% sodium hydroxide in a Stomacher Lab Blender. The resulting alkaline haematin extract was quantified spectrophotometrically at 564nm on Elisa plates. MBL was determined by comparing the alkaline haematin values with levels from similar extracts of whole blood standards (20-600 µl blood). Results were expressed as milliliters (ml) of blood lost. The alkaline haematin technique was highly reproducible with intraassay and interassay coefficient of variation estimates at 4.3 and 6.7, respectively (n=8 assays). MBL values were log transformed to correct for heterogeneity of variance and then subjected to ANOVA. The average total MBL during the menstrual cycle in 11 animals was 5.87 ± 0.6 ml (over 6 days). Menstruation peaked in all the animals on day 3 after P withdrawal. Short menstrual cycles had no significant effect on total MBL compared to normal-length cycles. However, treatment with artificially amplified secretory phases resulted in greater MBL (8.8 ± 2.2 ml/cycle). These animals had a 1 day delay of frank menses and then compensated with significantly heavier MBL on days 3, 4, and 5 after P withdrawal (P<0.05). We conclude that very accurate daily MBL measurements can be obtained from hormone controlled, artificially cycled rhesus macaques and that these animals can provide a useful animal model for evaluating factors that modulate menstrual bleeding. Supported by NIH grants HD18185 and RR000163. (poster)

Estrogen Reduces BDNF Level, but Maintains Dopaminergic Cell Density in the Striatum of MPTP Mouse Model

ABSTRACTDegeneration of dopaminergic (DA) axons in the striatum triggers upregulation of striatal trophic activity and striatal DA neuronal number in animal models of Parkinson's disease (PD). The present study investigated the effects of 17β-estradiol (E2) on brain-derived neurotrophic factor (BDNF) expression and the density of DA neurons in the striatum of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model in correlation with nigrostriatal DA innervation. Adult male C57Bl/6 mice were treated with E2 or vehicle for 11 days. Following 5 days of E2 or vehicle pretreatment, animals were injected with MPTP on day 6. On day 11, all mice were sacrificed, and the striatum were collected and processed for tyrosine hydroxylase (TH) and BDNF immunohistochemistry. Striatal TH-immunoreactive (IR) neurons were counted. Extent of DA innervation and BDNF expression in the striatum were assessed by measuring optical density of TH and BDNF immunoreactivity, respectively. Pretreatment with E2 partially prevented DA denervation and decreased striatal BDNF upregulation triggered by MPTP, but maintained the density of striatal TH-IR neurons to that observed in MPTP group. These findings suggest that estrogen protection of nigrostriatal DA axons against MPTP as well as preservation of the striatal TH-IR cell density in MPTP/E2 mice may be not mediated by BDNF.

Outcomes of medically-indicated (MED) oocyte cryopreservation (OC) cycles performed for fertility preservation (FP) compare favorably to oc cycles of infertile women who have completed a thaw cycle

OBJECTIVE: As more reproductive-age women are diagnosed with and survive cancer, OC use is increasing as a means of FP, particularly in those without a male partner. We compared outcomes of medically-indicated (MED) OC cycles to those of infertile women completing an OC thaw cycle (THAW), including their meiotic spindle evaluation (MSE). All MII oocytes cryopreserved in our lab undergo MSE 2 h post-harvest. DESIGN: Retrospective. MATERIALS AND METHODS: 31 MED (520 retrieved eggs) and 28 THAW (512 retrieved eggs) OC cycles were reviewed. Tumors in the MED group included 13 GYN, 10 breast, 3 leukemia, 2 colon, 2 CNS and 1 sarcoma. All patients underwent standard ovarian hyperstimulation followed by retrieval, MSE 2 h post-harvest, then OC of all MII oocytes. Both slow freezing and vitrification were used. 4 MED pts froze ½ as zygotes. RESULTS: In the THAW cycles, there were 16 (57%) pregnancies: 12 women delivered 17 liveborns (5 twins), 3 have ongoing gestations and 1 miscarried. 90% of eggs survived and there was a 94% pre-cryo to post-THAW MSE correlation. 2 MED thaw cycles resulted in 1 pregnancy. Cycle comparisons for THAW vs. MED are shown in the Table.TableComparison of medically-indicated (MED) OC cycles and OC cycles of infertile women who completed a thaw (THAW).THAWMEDPMean age at retrieval (y)31±1 (range 21-38)31±1 (range 20-40).9Mean E2 day hCG (pg/ml)3086±3001788±176<.001Mean no. eggs retrieved18±2 (range 6-61)17±2 (range 5-38).8Mean no. MII for OC14±211±1.1Mean no. MII with pre-cryo spindle10±19±1.08 Open table in a new tab CONCLUSIONS: The majority of oocytes in both groups were mature (MII) and exhibited a spindle. Women diagnosed with cancer can achieve adequate ovarian stimulation, egg production and MSE despite limited time to complete OC treatment and multiple outside stressors, including their underlying disease and its management. Based on our preliminary THAW data, we believe cancer pts should have a reasonable chance for pregnancy using their frozen eggs.

Laboratory evaluation in oocyte cryopreservation (OC) cycles suggests retrieved oocytes are comparable whether frozen for medical indications (MED), deferred reproduction (DR) or oocyte donation (OD)

OBJECTIVE: The use of OC is increasing. We know the human oocyte meiotic spindle (MS) is crucial to chromosome segregation and embryo development while the zona pellucida (ZP) aids in sperm binding and fertilization. Today, polarized microscopy (PM) affords non-invasive MS and ZP evaluation. All MII oocytes cryopreserved in our lab undergo PM MS and ZP assessment 2 h post-harvest. We compared OC cycle outcome in cases performed for MED vs. DR vs. OD. DESIGN: Retrospective. MATERIALS AND METHODS: All women were <40 y at time of OC. Tumors in the MED group included 13 GYN, 10 breast, 3 leukemia, 2 colon, 2 CNS & 1 sarcoma. All patients underwent standard ovarian stimulation followed by retrieval, PM for MS and ZP, then OC of MII oocytes. RESULTS: Cycle outcome is shown in the Table.TableCycle Comparison of MED vs. DR and OD.MED (n=31)DR (n=73)OD (n=8)Mean Age (y)31±1a (range 20-40)37±0.3b (range 29-40)31.1±1(range 24-37)Mean E2 day hCG1788±1761974±1253396±527Mean no. oocytes retrieved17±1 (range 5-38; total 520)14±1 (range 2-43; total 1056)19±3 (range 9-34; total 179)Mean no. MII eggs cryo'd11±1c (total 342)10±1d(total 515)15±3e (total 132)Mean no. eggs spindle positive9±1 (range 2-33)8±1 (range 0-30)13±3 (range 3-26)Mean MS retardance1.3±0.03 (range 0.5-2.3)1.5±0.2 (range 0.3-3.0)1.4±0.05 (range 0.6-3.0)Mean ZP optical density2.2±0.1 (range 0.7-6.6)1.9±0.02 (range 0.6-4.5)2.3±0.1 (range 0.7-4.1)MED relative to DR and OD, P<0.001:a-b,c-d; P=0.01: c-e. Open table in a new tab MED relative to DR and OD, P<0.001:a-b,c-d; P=0.01: c-e. CONCLUSIONS: The majority of retrieved oocytes in all groups were MII. Women diagnosed with malignancy achieved adequate ovarian stimulation, egg production and a similar number of spindle positive eggs compared to DR and OD patients, despite limited time for treatment and multiple outside stressors, including underlying disease. MS & ZP measurements were clinically comparable whether eggs were frozen for cancer, DR or OD.

Percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) for men with obstructive azoospermia: a review of 6 years

OBJECTIVE: For patients with confirmed diagnosis of obstructive azospermia (OA) there are two methods to retrieve spermatozoa for ICSI, percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE). In this study we assessed six years of ART outcomes for patients using epididymal or testicular sperm for their ICSI cycle.DESIGN: Retrospective analysis of 6 years of PESA/TESE procedures.MATERIALS AND METHODS: A total of 67 patients treated from January 2002 to December 2007, were evaluated. The patients were divided into Group I (PESA n=36) and Group II (TESE n=31). The ovarian stimulation protocols were not different between groups with majority consisting of late luteal phase GnRH-agonists or follicular phase GnRH antagonist. The data were statistically analyzed by chi-square and ANOVA.Table 1Group I vs Group IIFactorGroup IGroup IIP ValueFemale age31.8±4.230.±5.60.317FSH6.6±2.16.2±2.30.512LH5.07±4.064.3±1.60.322E242.7±28.940.2±25.80.714Plolactine17.3±9.219.2±10.50.458Male Age38.5±9.438.3±8.80.939Follicles>15 Day 107.4±4.28.5±5.40.377E2 day 102281±21612355±13300.881Day Oocyte retrieval12.5±1.0812.±.7650.643Total oocytes11.2±6.513.4±6.10.163IVF/ICSI9.03±510.9±4.90.128Fertilized Oocytes6.06±3.76.2±3.70.825Fertilization rates65.9±20.359.1±22.80.2Embryo transfer day2.8±.782.8±.710.78Number of embryo transferred2.8±.822.8±.710.96Number of Pregnancies20100.084Gestacional sacs.760±.89.5±.70.212Implantation rate26.80±30.118.30±30.40.265 Open table in a new tab The implantation rate was 27% and 17.6% respectively (p=0.15).The fertilization rate was significantly higher for the group I 67% versus 57.3% (p<0.01). The pregnancy rate, although higher in group I (55.5% versus 32.2%, respectively), was not statistically difference (p=0.08).CONCLUSIONS: Epididymal sperm from men with OA have a higher fertilization rate than testicular sperm and a trend for higher pregnancy and implantation rate. PESA should be preferred to TESE since has also additional advantages as low cost and minimal postoperative discomfort. OBJECTIVE: For patients with confirmed diagnosis of obstructive azospermia (OA) there are two methods to retrieve spermatozoa for ICSI, percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE). In this study we assessed six years of ART outcomes for patients using epididymal or testicular sperm for their ICSI cycle. DESIGN: Retrospective analysis of 6 years of PESA/TESE procedures. MATERIALS AND METHODS: A total of 67 patients treated from January 2002 to December 2007, were evaluated. The patients were divided into Group I (PESA n=36) and Group II (TESE n=31). The ovarian stimulation protocols were not different between groups with majority consisting of late luteal phase GnRH-agonists or follicular phase GnRH antagonist. The data were statistically analyzed by chi-square and ANOVA. The implantation rate was 27% and 17.6% respectively (p=0.15).The fertilization rate was significantly higher for the group I 67% versus 57.3% (p<0.01). The pregnancy rate, although higher in group I (55.5% versus 32.2%, respectively), was not statistically difference (p=0.08). CONCLUSIONS: Epididymal sperm from men with OA have a higher fertilization rate than testicular sperm and a trend for higher pregnancy and implantation rate. PESA should be preferred to TESE since has also additional advantages as low cost and minimal postoperative discomfort.

Effect of ovarian hormones on serum adiponectin and resistin concentrations

To investigate the effect of ovarian hormones on adiponectin and resistin levels in women. Experimental study. University hospital. Thirteen normally cycling women (7 in group 1 and 6 in group 2) and 8 postmenopausal women (group 3). Women of group 1 were investigated in a control cycle and in a subsequent cycle in which total abdominal hysterectomy plus bilateral salpingooophorectomy (TAH+BSO) was performed on day 3. In both cycles, the women received increasing doses of E(2) from days 3 to 5. Women of group 2 underwent TAH+BSO on day 3 without receiving any hormonal treatment. Women of group 3 received increasing doses of E(2) for 15 days. Adiponectin, resistin, and E(2) concentrations. In group 1, serum adiponectin and resistin levels did not show any significant changes for the week following day 3 and were similar in the two cycles. In group 2, adiponectin and resistin levels were similar before and after TAH+BSO and remained stable during the first postoperative week. In group 3, no significant changes in adiponectin and resistin levels were seen during the 15 days of E(2) administration. Adiponectin and resistin values were not affected either by estrogen treatment or after ovariectomy in women. It is suggested that ovarian hormones are not involved in the regulation of adiponectin and resistin secretion in women.

The Number of Small Antral Follicles (2-5 Mm) Determines the Ovarian Reserve

Age and antral follicle count (AFC) are widely used as markers of ovarian reserve in subfertile women. Ovarian reserve tests (FSH, Inhibin B, Clomiphene Citrate Challenge Test (CCCT), Exogenous FSH Ovarian Reserve Test (EFORT)) have been introduced to predict spontaneous pregnancy chances and ovarian response in IVF. The relation between the size of the antral follicles and the results of ovarian reserve tests is not extensively studied. We studied which size of antral follicles determines the response of the various ovarian reserve tests in a general subfertile population. Prospective cohort study, 1999-2003. Setting: outpatient fertility clinic of UMC Groningen and Martini Hospital, the Netherlands. Patients: subfertile women. Inclusion criteria: regular ovulatory cycles, subfertility for more than twelve months, at least one ovary and at least one patent tube. Exclusion criteria: severe male subfertility (a total motile sperm count < 1 x106) and endometriosis AFS III/IV. The number of antral follicles (sized 2-10 mm) and ovarian volume were measured by transvaginal ultrasound on cycle day three to five. Serum basal FSH and Inhibin B were measured, the CCCT was performed and a subgroup underwent the EFORT. CCCT: 100 mg clomiphenecitrate was applied from cycle day three till seven. The sum of serum FSH on day three and day ten is the outcome of the CCCT. EFORT: 300 IU subcutaneous injection of Puregon® was applied on day two of the consecutive cycle. Serum Estradiol (E2) was measured on cycle day two and 24 hours later. Delta-E2 is the result of E2 day three minus day two. Statistical analysis was performed by SPSS 11.0 for Windows. Non-parametric tests were used to determine Spearman’s correlations. A total of 490 subfertile women participated in this study. Mean age was 32.7 years (19.9-44.7), mean duration of subfertility was 26.4 months (12-153), mean cycle length was 28 days (21-42), 70% of the women had a primairy infertility. AFC was significantly correlated with ovarian volume (r=0.49; p<0.01), Inhibin B (r = 0.16; p<0.01), basal FSH (r=-0.31; p<0.01) and CCCT (r=-0.31; p<0.01). In the subgroup (n=62) the correlation of AFC with the EFORT was strong (r=0.54; p<0.01). The correlations after dividing the AFC in two groups (2-5 and 6-10mm) are shown in the table: Tabled 1Spearman’s coefficient * P-waarde<0,05 ** P-waarde<0,01 Spearman’s coefficient * P-waarde<0,05 ** P-waarde<0,01 The correlations of the number of small antral follicles (2-5mm) with basal FSH, CCCT and EFORT were statistically significant in contrary to the correlations of the larger follicles (6-10mm). Moreover the EFORT showed a strong correlation with AFC and can easily be used in clinical practice as a marker of ovarian reserve. The CCCT has no additional value to basal FSH. We conclude that the number of small follicles is responsible for the endocrine response and can be seen as a marker for the functional ovarian reserve.

What are determinants of the oocyte degeneration rate after intracytoplasimic sperm injection (ICSI)?

Background and Significance: Interest has evolved in determining the factors associated with oocyte degeneration after ICSI. Limited reports indicated that oocyte degeneration was associated with the ICSI technique. Objective: This purpose of this study was to determine whether oocyte degeneration was technician dependent in an experienced laboratory. Baseline characteristics and physician-adjustable ovarian stimulation variables were also evaluated. Design: Prospective study. Materials and Methods: Six thousand seven hundred eighty-nine intracytoplasmic injection procedures were analyzed to determine whether the oocyte degeneration rate was technician dependent. 230 independent, down-regulated cycles were examined to assess which predictors may be associated with the oocyte degeneration rate. Multivariate analyses were performed using the generalized linear model routines in Stata, Ver 7.0 (Stata Corporation, College Station, TX) to perform logistic regression allowing for clustering by cycle. The models were determined by backward elimination in a guided fashion, starting with a model including all the predictors (ICSI technician, stripping technician, baseline characteristics (age, day 3 FSH/ E2, AFC), physician-adjustable variables (start dose, total dose, number of mature oocytes retrieved, E2 on day of hCG, predicted E2/large follicle, number of 13–17mm (medium) and ≥18mm (large) follicles on the day of hCG, size of the lead follicle), then eliminating those that were not statistically significant one at a time guided by the previous literature and clinical experience. Tests were declared statistically significant for a two-sided p-value < 0.05. Results: Neither the ICSI technician nor the stripping technician was a determinant of the oocyte degeneration rate. However, the day 3 FSH, number of mature oocytes retrieved, and E2/large follicle were significant independent predictor of oocyte degeneration rate. The day 3 FSH was positively associated with the degeneration rate, and the number of mature oocytes retrieved and the predicted E2/large follicle were negatively associated with the degeneration rate. Physician-adjustable variables were not associated with the degeneration rate. Tabled 1numdegCoef.Std. Err.zP>|z|[95% Conf. Interval]Age.0065215.02394280.270.785−.0404056.0534486AFC.0019983.01526470.130.896−.0279199.0319165Day 3 E2.0036408.00231791.570.116−.0009023.0081838#MII Oocytes−.0335009.0142233−2.360.019−.0613781−.0056237Day 3 FSH.0437193.02188112.000.046.0008332.0866053Start Dose.0255645.14112290.180.856−.2510313.3021603Total Dose−.0081025.0112635−0.720.472−.0301787.0139736Pred E2−.0023453.0008457−2.770.006−.0040029−.0006878_cons−2.003593.8960944−2.240.025−3.759906−.2472804Pred E2 = E2 day of hCG/(#medium follicles * 0.75 + #large follicles) (Standard errors scaled using square root of Pearson X2-based dispersion). Open table in a new tab Pred E2 = E2 day of hCG/(#medium follicles * 0.75 + #large follicles) (Standard errors scaled using square root of Pearson X2-based dispersion). Conclusion: The ICSI or stripping technician did not impact the oocyte degeneration rate. The data suggests that oocyte degeneration is a function of oocyte aging and physicians cannot alter the inherent occyte degeneration rate within a down-regulated cycle.

Coordination of early antral follicles by luteal estradiol administration provides a basis for alternative controlled ovarian hyperstimulation regimens

ObjectiveTo investigate whether luteal E2 administration reduces size discrepancies of early antral follicles. DesignProspective, crossover study. SettingART unit, Clamart, France. Patient(s)Sixty women and 120 cycles. Intervention(s)On cycle day 3 (baseline day 3), all women underwent measurements of early antral follicles by ultrasound and serum FSH and ovarian hormones. From day 20 until the next cycle day 2, 30 of them received oral 17β-E2, whereas the remaining women served as controls. The day after E2 discontinuation (E2 day 3) or on subsequent cycle day 3 (control day 3), participants were reevaluated as on baseline day 3. Main outcome measure(s)Magnitude of follicular size discrepancies. Result(s)Follicular size discrepancies and follicular diameters were significantly attenuated on E2 day 3 (3.7 ± 0.5 mm) as compared with baseline day 3 (4.9 ± 1.0 mm), but not in controls (5.0 ± 0.8 vs. 4.9 ± 0.8 mm). FSH (4.3 ± 1.9 vs. 7.3 ± 3.3 mIU/mL) and inhibin B (34 ± 28 vs. 71 ± 32 pg/mL) levels were consistently lower on E2 day 3 than on baseline day 3 but remained unchanged in controls. Conclusion(s)Luteal E2 administration reduces the size and improves the homogeneity of early antral follicles on day 3. This approach may be instrumental in synchronizing follicular development during controlled ovarian hyperstimulation.

Improved controlled ovarian hyperstimulation in poor responder in vitro fertilization patients with a microdose follicle-stimulating hormone flare, growth hormone protocol

To assess the efficacy of a novel protocol-microdose GnRH agonist (GnRH-a), FSH, and GH, for the stimulation of IVF patients who were canceled previously on a standard luteal GnRH-a, FSH, GH protocol. Prospective evaluation using the patient's previous IVF stimulation attempt as historic controls. Private practice assisted reproductive technology center. Thirty-two patients who had prior ovulation induction cycles canceled using luteal phase GnRH-a suppression followed by exogenous gonadotropins and GH. Precycle treatment with oral contraceptives followed by follicular phase administration of 40 micrograms leuprolide acetate every 12 hours beginning on cycle day 3 and FSH supplemented with GH beginning on cycle day 5. Paired analysis of E2 day 5, number of follicles, ampules of FSH required, and cancellation rate. The number of oocytes, embryos, embryo quality, implantation rate, and pregnancy rate (PR) were determined for completed cycles on the microdose GnRH-a, FSH, GH protocol. Controlled ovarian hyperstimulation was superior during microdose GnRH-a, FSH, GH stimulation when compared with the prior luteal GnRH-a cycle. Specifically, there was a higher E2 response, more oocytes, fewer cycle cancellations, and no premature LH surge or luteinization. The microdose GnRH-a, FSH, GH protocol produced an average of 10 oocytes and a 50% ongoing PR. The microdose GnRH-a, FSH, GH protocol is superior to standard protocols for the treatment of patients with decreased ovarian reserve undergoing controlled ovarian hyperstimulation for IVF.

Ultrasonographic predictors of implantation after assisted reproduction

To assess the utility of endometrial thickness, echogenic endometrial pattern, and uterine artery impedance measured as pulsativity index in predicting implantation. Prospective case-controlled study of infertile patients undergoing assisted reproductive technologies (ARTs). Four hundred five women undergoing ARTs were studied: 100 women after ET of thawed embryos in natural cycles (frozen ET), 107 women after standardized IVF-ET, 99 women receiving donor oocytes after controlled endometrial development with estrogen and P, and 99 women undergoing IUI with various ovarian stimulation regimens (none, 16; GnRH, 7; clomiphene citrate [CC], 29; hMG, 47). Transvaginal ultrasonographic examination performed on the day of hCG administration during stimulated cycles; on E2 day 15 during controlled endometrial cycles; and on the day of ovulation during natural, CC, and GnRH pump cycles. The endometrial thickness, echogenic pattern, and pulsativity index results in 170 conception cycles were compared with 235 nonconception cycles. When conception and nonconception cycles were compared, no difference in mean endometrial thickness and significant differences in the frequency of nonmultilayered pattern and pulsativity index > 3.3 were observed. Forty-seven percent of nonconception and 9% of conception cycles were associated with at least one of these factors. Ultrasonic measurements of pulsativity index, resistance index, and echogenic pattern are useful in predicting implantation after assisted reproduction.