Edmond Confino

Promoter Methylation Regulates Estrogen Receptor 2 in Human Endometrium and Endometriosis

Abstract Steroid receptors in the stromal cells of endometrium and its disease counterpart tissue endometriosis play critical physiologic roles. We found that mRNA and protein levels of estrogen receptor 2 (ESR2) were strikingly higher, whereas levels of estrogen receptor 1 (ESR1), total progesterone receptor (PGR), and progesterone receptor B (PGR B) were significantly lower in endometriotic versus endometrial stromal cells. Because ESR2 displayed the most striking levels of differential expression between endometriotic and endometrial cells, and the mechanisms for this difference are unknown, we tested the hypothesis that alteration in DNA methylation is a mechanism responsible for severely increased ESR2 mRNA levels in endometriotic cells. We identified a CpG island occupying the promoter region (−197/+359) of the ESR2 gene. Bisulfite sequencing of this region showed significantly higher methylation in primary endometrial cells (n = 8 subjects) versus endometriotic cells (n = 8 subjects). The demethylating agent 5-aza-2′-deoxycytidine significantly increased ESR2 mRNA levels in endometrial cells. Mechanistically, we employed serial deletion mutants of the ESR2 promoter fused to the luciferase reporter gene and transiently transfected into both endometriotic and endometrial cells. We demonstrated that the critical region (−197/+372) that confers promoter activity also bears the CpG island, and the activity of the ESR2 promoter was strongly inactivated by in vitro methylation. Taken together, methylation of a CpG island at the ESR2 promoter region is a primary mechanism responsible for differential expression of ESR2 in endometriosis and endometrium. These findings may be applied to a number of areas ranging from diagnosis to the treatment of endometriosis.

Nonsurgical Fallopian Tube Recanalization for Treatment of Infertility

Fluoroscopic transcervical fallopian tube recanalization was performed in 100 consecutive patients with infertility and proximal tubal obstruction documented with hysterosalpingography. In 86 patients, the procedure enabled at least one tube to be opened. Twenty-six intrauterine pregnancies resulted from the successful recanalization. A well-defined subset of 20 patients were evaluated to better define the treatment effect of fallopian tube recanalization. All 20 had bilateral proximal tubal obstruction without other tubal disease, and all had been recommended for tubal microsurgery or in vitro fertilization. Recanalization of one or both tubes was successful in 19 of these women (95%). Nine patients conceived (47%) without receiving any other therapy, and the average time from procedure to conception was 4 months. All pregnancies were intrauterine. Eight of the 10 patients who did not conceive underwent follow-up hysterosalpingography an average of 6 months following the procedure; four (50%) demonstrated reocclusion of both tubes. The authors conclude that nonsurgical fallopian tube recanalization is an effective treatment for infertility caused by proximal tubal obstruction.

Histopathological Findings in 226 Women with Post-Menopausal Uterine Bleeding

Histopathological findings in 226 post‐meno‐pausal bleeding women were reviewed retrospectively. Ade‐nocarcinoma of the endometrium was diagnosed in 7% and hyperplastic endometrium in 15%. The incidence of malignancy showed a definite rise with advancing age, increasing amount and duration of bleeding, prolonged time interval between the menopause and onset of bleeding, and enlarged uterus. Adenocarcinoma of the endometrium was associated in 40% of the patients with either obesity, diabetes mellitus, or hypertension. The most frequent histopathological finding was atrophic endometrium (45%).

The predictive value of hCG β subunit levels in pregnancies achieved by in vitro fertilization and embryo transfer: an international collaborative study

Serial human chorionic gonadotropin (hCG) beta subunit measurements of 300 pregnancies achieved by in vitro fertilization (IVF) were obtained by 15 IVF centers worldwide. Hormonal curves were established for 164 normal singleton pregnancies, 25 normal multiple gestations, 60 chemical pregnancies, 41 first-trimester spontaneous abortions, and 10 ectopic pregnancies. In comparison to the normal singleton pregnancy curve, chemical pregnancies and spontaneous abortions showed statistically lower hCG levels. hCG levels of ectopic pregnancies, compared with normal singleton gestations, were lower from days 7 to 14. It is concluded that beta-hCG determinations of in vitro fertilized pregnancies allow pregnancy diagnosis as early as 7 to 9 days after embryo transfer (ET) and will permit early discrimination between normal and abnormal IVF pregnancies. A single beta-hCG determination on day 9 after ET may discriminate chemical, ectopic pregnancy or impending miscarriage from a normal gestation. An hCG determination on day 17 will predict early normal development of an IVF pregnancy.

Intrauterine inseminations with washed human spermatozoa

The success of intrauterine inseminations with washed human spermatozoa was evaluated in four groups of patients. The first two groups were couples in whom either the man or the woman had detectable circulating sperm-agglutinating antibodies. The third group included couples where the woman exhibited hostile or absent cervical mucus, whereas the fourth group represented couples with oligozoospermic males. The couples underwent 3.7 +/- 2.1 insemination cycles, with a 30% overall pregnancy rate for all four groups. Intrauterine inseminations of women with hostile cervical mucus yielded a 68% conception rate. Couples with either male or female serum sperm-agglutinating antibodies showed 25% and 40% pregnancy rates, respectively. However, none of the couples with oligozoospermic males achieved pregnancy. Intrauterine inseminations with washed human spermatozoa thus represent a satisfactory method of achieving pregnancy in women with hostile cervical mucus and in couples with circulating sperm agglutinating antibodies. Couples in whom oligozoospermia has been identified as the principal cause of infertility do not seem to benefit from this therapy.

Relationship between peak serum estradiol levels and treatment outcome in in vitro fertilization cycles after embryo transfer on day 3 or day 5

OBJECTIVE To examine the relationships between peak serum estradiol (E(2)) levels and treatment outcome in in vitro fertilization (IVF) cycles after embryo transfer (ET) on day 3 or day 5. DESIGN Retrospective analysis of 697 IVF-ET cycles between January 1999 and December 2001. SETTING A university-affiliated assisted reproduction program. PATIENT(S) Infertile patients undergoing IVF-ET cycles. INTERVENTION(S) Peak E(2) concentration in serum was determined on the day of human chorionic gonadotropin (hCG) administration. The IVF-generated embryos were cultured for 2 days until transfer on day 3. If more than four 8-cell embryos were present on day 3, embryo culture was continued until day 5 for blastocyst transfer. MAIN OUTCOME MEASURE(S) Clinical pregnancy rates. RESULT(S) High peak E(2) levels did not adversely affect treatment outcome. After the cycles were divided according to the day of ET, high peak E(2) levels were associated with improved pregnancy rates after ET on day 5 but not on day 3. CONCLUSION(S) Increasing peak E(2) levels in IVF cycles are associated with improved pregnancy rates after ET on day 5.

Transcervical balloon tuboplasty.

TBT is a new technique that by use of a balloon catheter technique in a fashion similar to that of balloon angioplasty allows reestablishment of tubal patency in selected cases of tubal occlusion. The case presented here describes the first transcervical dilatation and recanalization of a proximally occluded fallopian tube in a patient with infertility.

Peritoneal fluid and serum autoantibody levels in patients with endometriosis

Patients with endometriosis demonstrate autoantibody abnormalities in their peripheral blood. Whether such abnormalities can also be detected in the peritoneal cavity, has not been established. We therefore investigated autoantibodies to 6 phospholipid antigens, 5 histones and histone subfractions and 4 polynucleotides in 14 laparoscopically diagnosed endometriosis and 9 control patients, undergoing laparoscopic tubal occlusion, in both serum and peritoneal fluid. Endometriosis patients demonstrated significantly lower total immunoglobulin G (IgG) levels in peritoneal fluid than controls. In contrast, specific IgG autoantibody levels were higher in peritoneal fluids of endometriosis patients. Mean antiphospholipids and antihistones autoantibodies reached significance when serum/peritoneal fluid ratios were compared with controls. Significant differences between endometriosis and control patients were restricted to IgG isotypes and were not observed for either IgM or IgA isotypes. These data suggest an abnormal concentration of IgG antiphospholipids and antihistones antibodies within the peritoneal cavity of endometriosis patients, which may play a contributing role in the peritoneal pathology of this condition.

Steroidogenic factor-1 and endometriosis

Endometriosis is a common and chronic disease characterized by persistent pelvic pain and infertility. Estradiol is essential for growth and inflammation in endometriotic tissue. The complete cascade of steroidogenic proteins/enzymes including aromatase is present in endometriosis leading to de novo estradiol synthesis. PGE(2) induces the expression of the genes that encode these enzymes. Upon PGE(2) treatment, coordinate recruitment of the nuclear receptor SF-1 to the promoters of these steroidogenic genes is the key event for estradiol synthesis. SF-1 is the key factor determining that an endometriotic cell will respond to PGE(2) by increased estradiol formation. The presence of SF-1 in endometriosis and its absence in endometrium is determined primarily by the methylation of its promoter. The key steroidogenic enzyme in endometriosis is aromatase encoded by a single gene because its inhibition blocks all estradiol biosynthesis. Aromatase inhibitors diminish endometriotic implants and associated pain refractory to existing treatments in affected women.

Transcervical tubal cannulation, past, present, and future

OBJECTIVE To evaluate publications that introduced novel diagnostic and therapeutic transcervical procedures on the fallopian tubes. DESIGN Major studies that conceptually changed the therapeutic approach to the fallopian tubes were reviewed. Minor publications were also included if they introduced a new concept or contributed to the topic. Clinical publications were selected if they involved transcervical diagnosis and treatment of fallopian tubes. RESULTS Transcervical tubal catheterization procedures for diagnosis of tubal disease, tubal obliteration, tubal recanalization, and tubal medication are minimally invasive procedures that can improve our understanding and diagnostic accuracy of tubal disease. These procedures allow transcervical treatment of proximal tubal occlusion. Further improvements in equipment and methodology are promising. Transcervical tubal occlusion, gamete and embryo deposition, and treatment of ectopic pregnancy may all be performed using the transcervical approach. CONCLUSION Transcervical tubal catheterization can replace microsurgery and IVF in selected patients with proximal tubal occlusion, improve the diagnostic accuracy of tubal disease, and deliver medications to the fallopian tubes. Cumulative knowledge suggests that transcervical tubal catheterization should become a universally accepted, taught, and practiced approach in the diagnosis and treatment of the fallopian tubes.