The role of selective oestrogen receptor modulators in the treatment of endometrial bleeding in women using long-acting progestin contraception.

This paper explores the concept that endometrial breakthrough bleeding results from the stimulatory effects of oestrogen in the endometrium. Though 'progestin-only' contraceptive regimens have long been associated with user dissatisfaction because of unpredictable vaginal bleeding, it is likely that the substantial contribution of endogenous ovarian oestradiol during such treatments predisposes the bleeding problems. Oestrogen causes endometrial proliferation, hyperplasia and neoplasia if unopposed. Oestrogen allows production of growth factors supporting angiogenesis which results in an abundance of dilated or fragile endothelial surface blood vessels, predisposing this tissue to bleeding when these vessels lose competence.

A rapid immunoassay predicts BRCA1 and BRCA2 mutations in buccal cells.

We developed a rapid, non-invasive, and inexpensive, assay capable of identifying BRCA1 and BRCA2 (BRCA) mutations in buccal cells. To determine the predictive value of this immunoassay, a double blind study of 13 high risk individuals was conducted by two independent teams. As greater than 90% of BRCA mutations result in protein truncations, a diminished anti-carboxy immunoreactivity relative to anti-amino immunoreactivity was scored as predictive for mutation. Comparison to BRCA DNA analysis was undertaken. The positive and negative predictive values were 90% and 100% respectively (p<0.02), suggesting great promise as an inexpensive and rapid screen for BRCA mutations.

Immunohistochemical analysis of BRCA2 expression in normal human buccal cells.

Previously we demonstrated that protein coded by the BRCA1 gene was expressed in normal human buccal cells. The present study confirmed that BRCA2 protein was similarly expressed in these cells. Messenger RNA for BRCA2 was detected with sequential use of two primer sets. Pooled cell samples from healthy donors reacted strongly with two commercially available antibodies, I17 and C15. Immunoreactivity was present in both cytoplasmic and nuclear compartments. We conclude buccal cells will provide a suitable model for exploration of normal BRCA function.

An antibody assay predictive of BRCA1 mutations in ovarian tumors and normal tissue.

To determine whether immunohistochemistry can identify BRCA1 mutations, immunohistochemical (IH) analysis was undertaken on paraffin sections of paired ovarian cancer and normal tissue using antibodies against both terminal regions of the BRCA1 protein. Ten patients at risk for BRCA1 mutations were studied. The results of BRCA1 mutation analysis and IH were compared. In tumor, IH correctly identified the presence or absence of loss of heterozygosity in all specimens. In all uninvolved specimens, IH correctly identified the presence or absence of a germline mutation. The sensitivity, specificity, positive and negative predictive values were 100% suggesting promise as a rapid and inexpensive screen.

Immunohistochemical analysis of BRCA1 expression in normal human buccal cells.

While tumor suppressor properties of BRCA1 are well documented, less is understood concerning the normal function of this protein in healthy adult tissues. We demonstrate here that BRCA1 protein is expressed in human buccal cells. Expression of BRCA1 mRNA transcripts in buccal cells was confirmed by PCR of reverse transcribed RNA. Three BRCA1 antibodies, D20, MS110 and MS13, reacted positively with cytospin deposited buccal cells, demonstrating apparent cytoplasmic and nuclear distribution of BRCA1 protein. Double antibody immunofluorescent, binding of D20 and MS110 was differentially distributed. We conclude that buccal cells provide a cell source for exploration of BRCA1 protein function in normal human adults.

Chronic antiprogestin therapy produces a stable atrophic endometrium with decreased fibroblast growth factor: a 1-year primate study on contraception and amenorrhea.

OBJECTIVE: To describe the efficacy of mifepristone in the prevention of menstrual bleeding and ovulation, with similar observations in comparison groups. DESIGN: Prospective experimental study. Thirty-two cynomolgus monkeys were divided equally into four treatment groups (n = 8). Treatment lasted for 1 year. INTERVENTION(S): Group I received GnRH-agonist (GnRH-a) and in-sequence mifepristone, group II received mifepristone only, group III received GnRH-a only, and group IV received vehicle control. MAIN OUTCOME MEASURE(S): Serum estradiol and progesterone, menstrual bleeding, endometrial thickness, and endometrial expression of basic fibroblast growth factor (bFGF) as determined by immunohistochemistry. RESULT(S): Weekly progesterone determinations showed that mifepristone-treated monkeys seldom ovulated (6 ovulations in 8 years), compared with the controls (100 ovulations in 8 years), while maintaining early to midfollicular levels of circulating serum estradiol. The GnRH-a-only group also rarely ovulated, but was chronically and severely hypoestrogenic. The mifepristone-only group showed scant menstrual bleeding (5 days in 8 years) as compared with the menstrual frequency in control animals (422 days in 8 years). Endometrial proliferation, as determined by biopsy, was similarly minimal for both the GnRH-a and mifepristone groups, and statistically less than in control monkeys. Both the mifepristone and GnRH-a treatments suppressed endometrial gland expression of the angiogenesis polypeptide bFGF. CONCLUSION(S): Chronic mifepristone induced anovulation along with virtual amenorrhea, which suggests the worth of this novel hormonal contraceptive.

Pelvic adhesions contain sex steroid receptors and produce angiogenesis growth factors.

OBJECTIVE: To evaluate female pelvic adhesion tissue for the presence of estrogen receptor (ER), progesterone receptor (PR), basic fibroblastic growth factor (basic-FGF), and vascular endothelial growth factor (VEGF). DESIGN: Descriptive study. SETTING: Patients at a tertiary medical center. PATIENTS: Female reproductive age patients undergoing gynecologic surgery who were not receiving hormonal therapy. INTERVENTIONS: Female reproductive tract peritoneal adhesion tissue was excised, frozen, and sent for immunohistologic evaluation. MAIN OUTCOME MEASURE: Presence of ER, PR, basic-FGF, and VEGF in adhesion tissue. RESULTS: Nineteen of 19 specimens were positive for PR; 16 of 19 specimens were positive for ER, which was present in a variety of the different cell types constituting adhesion. Vascular endothelial growth factor and basic-FGF were detected in endothelial cells of blood vessels supplying this tissue as well as in mesothelial cells. CONCLUSION: Adhesion tissue contains ER, PR, and growth factors that may be important in the genesis of the permanent fibrovascular bands between pelvic organs. This supports the theoretical possibility of hormonal manipulation of these tissues to negatively influence postoperative pelvic adhesion formation.