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Biomedical subjects

E Radwanska

Publications and source records attributed to E Radwanska.

At least 19 recordsLinked to original sources

Evidence for different aetiologies of low estradiol response to FSH: age-related accelerated luteinization of follicles or presence of ovarian autoantibodies.

BACKGROUND: We found granulosa cells of low responders (LR) expressed more LH receptors, suggesting that follicles were more luteinized than normal responders (NR). The objectives were to test the hypothesis that follicles of LR were more luteinized than follicles of NR, and to determine if LR with (LR+) and without (LR-) ovarian antibodies differed. METHODS: Hormone levels and ovarian autoantibodies (OVAB) were measured in follicular fluid from mature follicles (>17 mm), and in serum obtained on the day of oocyte retrieval during controlled ovarian stimulation. The gonadotrophin response was defined as a ratio of peak estradiol/number of FSH ampoules. RESULTS: NR (32.5 +/- 4.6 years; n = 11) were similar in age to LR+ (33.4 +/- 4.2 years; n = 9) and were younger than LR- (37.1 +/- 3.8 years; n = 12) (P = 0.03). Likewise, dehydroepiandrosterone sulphate was lower in LR- compared with LR+ or NR (P < 0.01). FSH, progesterone, inhibin-A and vascular endothelial growth factor levels were higher in follicular fluid of LR than NR. LR- and LR+ differed. For example, the follicular fluid progesterone/estradiol ratio was similar in NR (11.1 +/- 8.9) and LR+ (9.8 +/- 6.6) but was lower than LR- (22.9 +/- 19.6) (P = 0.05). Serum hormone levels did not reflect follicular fluid hormone profiles. CONCLUSIONS: In the absence of ovarian antibodies, low responses are associated with higher age and accelerated luteinization of mature follicles, rather than diminished responsiveness. Ovarian antibody may be an additional tool to predict and individualize treatment regimens in poor responders.

Adult↗

Effects of dioxin, an environmental pollutant, on mouse blastocyst development and apoptosis.

OBJECTIVE: To evaluate the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; dioxin) on mouse embryo development and apoptosis. DESIGN: Controlled animal study. SETTING: Academic research environment. ANIMAL(S): Female mice (CB6F1) at 3 to 6 weeks of age and proven breeders (C578B46). INTERVENTION(S): Mouse embryos were obtained at the morula stage and cultured to the blastocyst stage in a pharmacologic dose of TCDD (3.1 microM) or a control medium. The morphology was assessed, and staining for apoptosis was performed. Immunohistochemistry for the presence of aromatic hydrocarbon receptor (AhR) was performed in another set of morula-stage embryos. MAIN OUTCOME MEASURE(S): The number of embryos developing from the morula to the blastocyst stage and number of apoptotic blastomeres in control vs. TCDD culture conditions. RESULT(S): No statistically significant differences were observed in the percentage of embryos reaching the blastocyst stage: 80.9% (115 of 142) in the TCDD-treated group, vs. 82.9% (121 of 146) in the control group. There was also no difference in the degree of apoptosis: 22.6 +/- 7.3% apoptotic cells (TCDD) vs. 25.3 +/- 9.7% (controls). Staining indicated the slight presence of aromatic hydrocarbon receptor in the morula-stage mouse embryos. CONCLUSION(S): TCDD at 3.1 microM did not alter the development of early mouse morula to blastocysts and did not significantly induce apoptosis in vitro.

Animals↗

Gonadotrophin receptor expression on human granulosa cells of low and normal responders to FSH.

The aim of this study was to determine if follicle stimulating hormone receptor and luteinizing hormone receptor (FSH-R and LH-R) expression is altered on granulosa cells (GC) of women with low oestradiol responses to FSH. Cells were obtained from mature follicles (>17 mm) following controlled ovarian stimulation. For comparison, chinese hamster ovary (CHO) cells transfected with FSH-R or LH-R were also assessed. FSH-R and LH-R expression were detected by flow cytometry. Receptors were labelled with FSH-R antibodies, or with excess FSH or human chorionic gonadotrophin (HCG) and anti-FSH or HCG antibodies, and compared to multiple controls. Receptor expression on GCs was more heterogeneous than on CHO cells. Gonadotrophin receptor levels on GCs were not correlated with the number of FSH ampoules administered or peak oestradiol response. Low and normal response groups were defined using a ratio of peak oestradiol/number of FSH ampoules. FSH receptor expression was not different on GCs from low and normal responders. However, LH-R expression was higher on GCs of low responders compared to those of normal responders (P = 0.04 ) suggesting a trend to more advanced luteinization. Access of hormone to follicles was not reduced in low responders. Thus, differences in gonadotrophin receptor expression, hormone binding, and access of hormones to follicles do not appear to account for low oestradiol responses to FSH.

Adult↗

Ovarian autoimmunity: greater frequency of autoantibodies in premature menopause and unexplained infertility than in the general population.

The objective of this study was to: (1) assess the relative prevalence of ovarian, thyroid, nuclear, and cardiolipin antibodies associated with premature menopause and unexplained infertility and (2) compare ovarian and thyroid antibodies in premature menopause, unexplained infertility, and the general population. Autoantibodies were evaluated in women with premature menopause (n = 30), unexplained infertility with (n = 38) or without (n = 15) prior gonadotropin-induced ovulation, and normal cycling controls (n = 12) and in a population of women obtained from a blood bank (n = 53). Antibodies to ovary (OVAB), thyroid (THYAB; thyroid peroxidase and thyroglobulin), cardiolipin, and eight nuclear antigens were assessed by enzyme immunoassay. Organ-specific antibodies (ovary and thyroid) were present with significantly greater frequency than non-organ-specific antibodies (nuclear and cardiolipin) in premature menopause and unexplained infertility (60% (50/83) vs 16% (13/83) respectively; P < 0.0001). OVAB (53%, 44/83) were significantly more frequent than THYAB (30%, 25/83) in premature menopause and unexplained infertility (P = 0.0030). THYAB did not differ among all groups (P = 0.78). In premature menopause and treated or untreated unexplained infertility OVAB frequencies were 53, 61, and 33%, respectively, and were significantly more frequent than in the population (17%) (P = 0.0001). In unexplained infertility, individuals with no prior gonadotropin-induced ovulation had a lower frequency of OVAB than treated individuals (P = 0.07). The frequency distribution of optical density values for OVAB was significantly higher for premature menopause and unexplained infertility than for population or normal cycling women (P < 0.0001). Thus, only ovarian antibodies were significantly more frequent than other antibody markers of autoimmunity in premature menopause and unexplained infertility.

Adult↗

The relationship of human granulosa-lutein cell proliferative index to follicular diameter and serum estradiol.

OBJECTIVE: To examine the relationships of follicular diameter and serum estradiol (E2) to the percentage of granulosa cells undergoing mitosis as reflected by the proliferative index of granulosa cells. METHODS: In this prospective study, we enrolled 44 consecutive women undergoing controlled ovarian hyperstimulation for in vitro fertilization. Deoxyribonucleic acid histograms were generated by flow cytometry from granulosa cells isolated at the time of transvaginal aspiration. Proliferative index was defined as the sum of G2/M and S phases. We assessed the correlation between proliferative index and age, maximum serum E2, number of oocytes retrieved, percent mature oocytes, and follicular diameter. RESULTS: Follicles less than 16 mm had a significantly higher proliferative index (19.9 +/- 3.3%) than follicles 20 mm or greater (14.8 +/- 3.9%, P = .016). However, there was no significant difference between proliferative index of the latter group and proliferative index of follicles 16-19 mm (17.8 +/- 4.7%). An inverse correlation between patient age and proliferative index of granulosa cells was noted (r = -.39, P = .018). There was no significant relationship between serum E2 and proliferative index (P = .97). CONCLUSION: Mitotic activity tends to decrease as follicular diameter increases after a threshold diameter is achieved. Proliferative index of granulosa cells provides insight into the underlying cell biology of a follicle.

Adult↗

The effect of the oviduct, uterine, and in vitro environments on zona thinning in the mouse embryo.

OBJECTIVE: To evaluate the impact of the oviduct, uterine, and in vitro environments on zona pellucida thinning in the mouse embryo. DESIGN: Female mice were stimulated with pregnant mare serum gonadotropin and mated and hCG injection. Unilateral oviduct ligation was performed on day 2 of gestation using the dorsal approach. The mice were divided into equal groups and killed on days 2, 3, 4, 5, and 10 of gestation. In vitro incubated embryos served as controls. Average daily zona thickness measurements were subjected to analysis of variance and paired Student's t-test. SETTING: The laboratory of the assisted reproductive program of Rush University Medical Center. MAIN OUTCOME MEASURE(S): Progressive daily decrease in average zona thickness. RESULT(S): Zona measurements of embryos flushed out of uterine horns, ligated oviducts, and in vitro incubation demonstrated statistically significant decreases in zona thickness, from 9.6 +/- 1.6 microns (day 3) to 6.0 +/- 0.8 microns (day 5), from 11.6 +/- 2.2 microns (day 2) to 6.0 +/- 1.6 microns (day 5), and from 11.1 +/- 2.0 microns (day 2) to 6.0 +/- 1.6 microns (day 5), respectively. There were no differences in average zona thickness for embryos in the same cell stage and same protocol day in all three locations. CONCLUSION(S): Zona thinning seems to be induced primarily by the dividing embryo before implantation. A substantial tubal and uterine contribution to zona thinning was not detected in this mouse embryo model.

Animals↗

Oocyte maturation in rabbits: effects of calmodulin inhibitors.

Oocyte maturation in mammals follows a highly conserved pattern of release from arrest through to the extrusion of the first polar body and formation of the second metaphase spindle. Oscillations in cytoplasmic calcium concentration precede the events of maturation in many species. These calcium ions interact with and activate calcium-binding proteins, including calmodulin, within the cell. Thus, it was of interest to us to examine whether calcium acted through calmodulin in the initial stages of maturation in rabbit oocytes or whether calmodulin was required for continuation through metaphase I no to metaphase II. Using the calmodulin inhibitor W-7 we found a significant (p < 0.05) decrease in the percentage of oocytes that underwent germinal vesicle breakdown. Calmidazolium did not prevent germinal vesicle breakdown; however, it caused a significant (p < 0.05) decrease in the proportion of oocytes with fully elaborated spindles and taxol-induced cytoplasmic asters. Both inhibitors caused a significant (p < 0.05) reduction in the proportion of oocytes that extruded their first polar bodies. The kinase inhibitor 6-DMAP caused a significant reduction in the proportion of oocytes with spindles and condensed chromatin, indicating the necessity for phosphorylation events in the resumption of meiosis. In rabbit oocytes calmodulin may play a role in the release from prophase arrest, and it is necessary for spindle preservation and continuation through metaphase I to metaphase II. The varying effects of the two inhibitor stems from their different binding sites on the calmodulin molecule thus causing a differential effect on its downstream effectors.

Adenine↗

Optimizing tubal epithelial cell growth promotes mouse embryo hatching in coculture.

PURPOSE: This study investigates the relationship between human tubal epithelial cell growth characteristics and mouse embryonic development to determine which cellular requirements should be preferentially provided in a coculture system. METHODS: Cell growth and viability were assessed for 5 days in alpha-minimal essential medium or human tubal fluid supplemented with 10% human serum or 10% synthetic serum. Two-cell mouse embryo development to blastocyst and hatching blastocyst stages was also assessed with or without coculture. RESULTS: Both epithelial cell growth and embryo development were dependent on serum supplementation with better cell viability and growth rates in human serum and better blastocyst development in synthetic serum. The highest proportion of hatching blastocysts was found in alpha-minimal essential medium and human serum with coculture. CONCLUSIONS: Culture conditions which improve tubal epithelial cell growth also improve the hatching rate of mouse embryos in coculture. This indicates that by meeting the metabolic and nutritional demands for epithelial cell growth, the beneficial effects of coculture on embryo development may be optimized.

Animals↗

Morphologic changes in offspring of female mice exposed to ethanol before conception.

OBJECTIVE: Our purpose was to evaluate the mutagenic effects of preconceptional ethanol exposure by use of the mouse model. STUDY DESIGN: Fifty-three adult female CF-1 mice were divided into one of five groups with or without an ethanol diet for 28 days. Mice were then superovulated and mated. On day 14 of gestation all mice were killed and fetuses removed and examined for abnormalities. RESULTS: The mutation index for the study and mutagen groups was similar (48% vs 41%). The percentage of any anatomic abnormality in any treatment group were between 50% and 100%. Anomalies of the abdominal wall were most commonly seen in the mutagenic groups. CONCLUSIONS: Chronic ethanol exposure in the female mouse before conception yields anatomic abnormalities in the offspring. These mutagenic effects may be variable in that they may result in a dominant lethal mutation or a delayed interference with organogenesis. Ethanol consumption, whether before or after conception, poses an unfavorable outcome in the offspring produced.

Abnormalities, Drug-Induced↗

Comparison of mouse embryo development in open and microdrop co-culture systems.

Co-culture with numerous cell lines has been shown to improve in-vitro embryo development. It is usually performed in open culture without an oil overlay, or in relatively large volumes of medium (e.g. 0.5 ml) under oil. We compared the efficacy of open and microdrop co-culture systems using human endometrial and tubal cell lines and mouse zygotes. Although the mean pH values of the media from the tubal cell cultures (both open and oil-covered) decreased significantly over 5 days of culture, this did not appear to impair embryo development. Both co-culture and microdrop culture significantly improved blastocyst and hatching blastocyst formation rates. The combination of the two techniques (microdrop and co-culture) demonstrated the highest blastocyst formation and hatching blastocyst formation rates, as well as the highest mean cell numbers in hatching blastocysts. Co-culture in a microdrop is a superior system for mouse embryo culture.

Animals↗

Sonographically monitored ovarian stimulation for assisted reproduction: a prospective, blind study.

OBJECTIVE: To assess prospectively the appropriateness of follicular sonography alone for monitoring ovarian stimulation and to compare it to ovarian monitoring with both follicular sonography and hormone level determinations. STUDY DESIGN: Prospective, blind, clinical study in which the investigator made cycle management decisions based on follicular sonography only. RESULTS: Follicular sonography alone predicted 88% of the decisions made by the combination of follicular sonography, luteinizing hormone (LH), estradiol (E2) and progesterone measurements. Follicular sonography was unable to predict abnormal E2 patterns in eight (8%) of the patients' scans. Follicular sonography did not detect three (3%) patients with a premature LH surge. CONCLUSION: Follicular sonograms alone performed during ovarian stimulation predicted 88% of cycle decisions. One could argue that hormone measurements could be either reduced or eliminated during ovarian stimulation for assisted reproductive technology and that follicular sonography only would be a cost-effective compromise. The effect of such simplified monitoring on pregnancy rates would require further prospective evaluation.

Female↗

Controlled ovarian hyperstimulation regimens in assisted reproductive technologies.

The goal of assisted reproductive technologies (ART) is to influence the recruitment of multiple, mature ovarian follicles. Several methods, including spontaneous cycle ART, clomiphene-based ART regimens, and gonadotropin regimens with and without adjuncts, are used. The controversies surrounding these techniques and their relative advantages and drawbacks are reviewed.

Female↗

Effects of induced hyperprolactinemia on in vitro fertilization cycles.

OBJECTIVE: To investigate the effect of induced endogenous hyperprolactinemia on the luteinization process, as expressed by the shift in the P:E2 ratio after hCG injection in IVF cycles. PATIENTS AND INTERVENTIONS: Serum PRL, E2, and P levels were measured in 49 IVF patients (leuprolide acetate and hMG protocol) on the day of hCG injection. Estradiol and P also were measured on the day after hCG. Serum P:E2 ratios were calculated for two groups of patients; group I (control): PRL < or = 20 ng/mL (conversion factor to SI unit, 1.00); group II (hyperprolactinemia): PRL > 20 ng/mL. Estradiol and P also were measured in follicular fluid (FF) and the gamete performance was compared between groups. RESULTS: Data analysis showed no significant differences in the mean +/- SD serum peak E2 (pg/mL; conversion factor to SI unit, 3.671) between groups: group I, 1,769 +/- 843; group II, 2,333 +/- 1,194; the mean FF E2 (pg/mL) group I, 351 +/- 221; group II, 370 +/- 186; or the mean FF P (ng/mL; conversion factor to SI unit, 3.180) group I, 8,357 +/- 3,127; group II, 11,354 +/- 12,888. No significant differences were found between groups in the P:E2 ratios on days 1 or 2: group I, 78 +/- 48 and 209 +/- 137; group II, 70 +/- 47 and 224 +/- 197, respectively. The magnitude of the P shift also showed no significant difference between the two groups; the mean +/- SD shift in the P level was 2.9 +/- 2.2 for group I, and 4.3 +/- 5.1 for group II. The serum PRL level had no effect on the fertilization rate (60% for group I and 70% for group II) or on the pregnancy rate (17% for group I and 23% for group II). CONCLUSION: These findings suggest that mild endogenous hyperprolactinemia induced by ovarian stimulation does not affect granulosa cell luteinization and gamete performance in humans.

Chorionic Gonadotropin↗

Evaluation of co-culture and alternative culture systems for promoting in-vitro development of mouse embryos.

The goal of this study was to compare mouse embryo development in a defined synthetic medium (human tubal fluid) against the same medium supplemented with a defined synthetic serum (SS), co-culture on human tubal epithelium (TECC), and culture on human fibronectin (FN) with and without SS. After 48 h, TECC, SS and FN + SS cultures demonstrated accelerated development with > 70% achieving > or = 8-cell stage. After 72 h, these culture conditions also significantly increased the proportion of embryos reaching the blastocyst stage but only TECC significantly increased the number of hatching blastocysts. Nuclei of the trophectoderm of unhatched and hatched blastocysts were stained with propidium iodide before fixing and labelling both the trophectoderm and inner cell mass with bisbenzimide. Blastocysts from the TECC contained a significantly higher total cell number (TCN) and trophectoderm and inner cell mass cell numbers than all other groups. These findings indicate equivalent improvements in mouse embryo development to the blastocyst stage in response to TECC, SS and FN and an enhanced number of cells and rate of hatching found only with TECC.

Animals↗

Luteal phase estradiol and pregnancy outcome in gonadotropin releasing hormone agonist/human menopausal gonadotropin-treated gamete intrafallopian transfer cycles.

To correlate luteal estradiol (E2) levels with pregnancy outcome, 36 consecutive conceptions resulting from gamete intrafallopian transfer in gonadotropin releasing hormone agonist/human menopausal gonadotropin (GnRH-a/hMG) cycles were analyzed. GnRH-a was initiated during the preceding luteal phase. HMG was adjusted individually. Human chorionic gonadotropin (hCG), 5,000 IU, was administered when E2 was > 500 pg/mL and the leading follicle > 17 mm (day 0). The luteal phase was supported by (1) hCG, 1,500 IU in three doses from day 5 and (2) progesterone (P) from day 7. E2 and P levels were analyzed in three groups of patients: normally progressing pregnancy (NPP), abortion (AB) and preclinical abortion (PAB). No significant differences in mean E2 levels were seen between the groups from day 0 through day 5 after hCG. Midluteal E2 levels were significantly different between the groups (P < .05). Late luteal E2 values were significantly higher for NPP than for either AB or PAB (P < .05). There were no significant differences in luteal P values between the NPP, AB and PAB groups. Decreased luteal E2 appears to be associated with early pregnancy wastage; this may be due to inadequate endometrial support.

Abortion, Spontaneous↗

The relationship between total motile sperm count and the success of intrauterine insemination.

OBJECTIVES: To determine if the total motile sperm count is predictive of IUI success and to apply life-table analysis to a large database of IUI procedures. DESIGN: Contingency table and life-table analyses were used to analyze a large group of couples (n = 546) undergoing 1,205 cycles of therapeutic IUI with husband's sperm. SETTING: Clinical infertility practice at a medical school referral center. PATIENTS: Five hundred forty-six couples with varying etiologies of infertility. RESULTS: There was a trend toward an increasing percentage of conception with increasing total motile sperm count. Significance was reached when the total motile sperm count used for IUI exceeded 20 x 10(6). The cumulative pregnancy rate calculated from this data reached 52% after the sixth attempted cycle of IUI.

Adult↗