The SCID mouse: an experimental model for endometriosis.

The purpose of this study was to validate the suitability of the severe combined immunodeficient (SCID) mouse as an experimental model for endometriosis, by defining the morphological and histological features of induced endometrial implants, and characterizing specific biochemical properties of these implants. Human secretory endometrial tissues were injected into the peritoneal cavity of SCID/SCID CB17 mature female mice. Successful peritoneal implantation was observed in 55 of 57 (96.5%) SCID mice and consisted of circumscribed elevated nodules. Haematoxylin-eosin staining of implanting lesions demonstrated the presence of endometrial glandular tissue in a mixed background of stromal and inflammatory cells. When progesterone was administered to mice, epithelial glands underwent well-defined secretory changes. Immunohistochemical analysis using polyclonal human pan-cytokeratin antibodies demonstrated selective positive staining in the glandular epithelium of the human implants with none in the surrounding stroma. In-situ hybridization analysis using complement component 3 cDNA radiolabelled riboprobes yielded significantly more intense signals in glands compared to stroma. As human endometrial implants in SCID mice were shown to retain specific histological, functional and biochemical properties, we conclude that the SCID mouse is an attractive animal model for the study of endometriosis.

Impact of hormone replacement therapy on the body mass and fat compositions of menopausal women: a cross-sectional study.

OBJECTIVE: The purpose of the study was to compare the body mass and fat compositions of menopausal women who were taking conventional doses of hormone replacement therapy (HRT) with that of menopausal women who were not taking any hormones. DESIGN: The body fat composition of 169 healthy postmenopausal women was measured using a noninvasive handheld machine, the Electrolipograph (BioAnalogics ELG, Beaverton, OR, USA). Impedance to electrical flow in tissues is lower with increasing water content of the tissue. Information on HRT, lifestyle, diet, smoking, and alcohol was obtained from the medical record and by a telephone interview before women were invited to participate. HRT and non-HRT groups were compared. Multivariate linear regression, which included age, years since menopause, type of menopause, and use of HRT, was performed for each of the two major outcomes: body mass index (BMI) and percentage of body fat. RESULTS: Comparisons between subgroups showed a large number of significant differences reflecting differences in age since menopause, baseline BMIs, and baseline waist to hip ratios. In the regression model, however, the only factor significantly associated with lower fat and BMI was the use of HRT. Women who were taking HRT had significantly lower percentages of body fat (-4.8%; p < 0.001) and BMI (-2.6 kg/m2; p < 0.001) compared with nonusers. Age and duration and type of menopause were not significant predictors of weight and BMI in this group of postmenopausal women. CONCLUSIONS: In this study, HRT seems to be associated with a significant reduction in postmenopausal weight and fat mass gains. This may be an important mechanism by which HRT exerts its beneficial long-term effects on cardiovascular health.

Masculinizing granulosa cell tumor of the ovary in a postmenopausal woman. A case report.

BACKGROUND: A total of 32 cases of virilizing granulosa cell tumors of the ovary have been reported. The current case has some unique features not previously reported. CASE: A 78-year-old woman presented with symptoms and signs of masculinization. A large, painless abdominal mass was discovered. Exploration revealed the mass to be originating in the left ovary, and surgical resection resulted in prompt reversal of the clinical and biochemical hyperandrogenic manifestations. Morphologic studies demonstrated a homogeneous granulosa cell tumor. CONCLUSION: This is the oldest patient on record with a masculinizing granulosa cell tumor and also the only masculinizing tumor presenting with advanced, stage III disease. Such tumors, although rare, should be considered in the differential diagnosis in postmenopausal women presenting with masculinizing symptoms.

Elevated expression of the proapoptotic BCL-2 family member, BAK, in the human endometrium coincident with apoptosis during the secretory phase of the cycle.

OBJECTIVE: To characterize the distribution of BAK (BCL-2 homologous antagonist/killer) protein in the human endometrium relative to the occurrence of apoptosis. DESIGN: A prospective, controlled study. SETTING: An academic hospital. PATIENT(S): Premenopausal women with histologically normal endometrium who were undergoing hysterectomy and curettage. INTERVENTION(S): Endometrial tissues were collected. MAIN OUTCOME MEASURE(S): Detection of BAK protein by immunohistochemical and immunoblot analyses, and of apoptosis by in situ DNA end-labeling. RESULT(S): BAK protein was detected in secretory endometrium and was confined to the glandular epithelial cells of the functionalis layer. Immunoreactive BAK was absent from most of the cells of the proliferative endometrium. By immunoblot analysis, we confirmed the immunohistochemical data by demonstrating that a 30-kD protein corresponding to BAK was elevated in lysates prepared from secretory, as compared with proliferative, endometrium. The elevated levels of BAK coincided with the onset of apoptosis in endometrial glandular epithelial cells during the secretory phase of the menstrual cycle. CONCLUSION(S): BAK protein localizes to glandular epithelial cells on the verge of apoptosis in the human endometrium. Thus, BAK likely functions with other members of the BCL-2 family in the regulation of apoptosis in the human uterus.

Endometrial alpha-2 macroglobulin; localization by in situ hybridization and effect on mouse embryo development in vitro.

alpha-2 macroglobulin (A2M) is a 718,000-kDA broad spectrum plasma protease inhibitor whose production by the human endometrium was recently reported. The multifunctional A2M receptor, also known as low-density lipoprotein receptor-related protein, was also recently immunolocalized to the endometrial stroma. The objective of this study was to further characterize the endometrial site of expression of A2M, and to study its effects on mouse embryo development in vitro, to gain some insight into the functional significance of its endometrial production. Formalin-fixed, paraffin-embedded human endometrium from hysterectomy and endometrial biopsy specimen was used for in situ hybridization analysis, with 35S-labeled riboprobes representing subcloned A2M complementary DNA (cDNA) fragments. Duplicate sections of human endometrium were hybridized with sense and antisense probe and coated with photographic emulsion. Resultant autoradiograms were analyzed qualitatively by light- and darkfield microscopy and quantitatively by a computerized analysis of the signal intensity. Immunohistochemistry and immunoblotting for endometrial tissues were performed using an affinity-purified polyclonal antibody to human A2M. The effect of A2M on mouse embryo development was studied by exposure of one cell mouse embryo in culture to physiological concentrations of biologically active and inactive A2M. Expression signals for A2M were more numerous and intense in the secretory endometrium, compared with proliferative endometrium. Endothelial cells lining the endometrial blood vessels seemed to be the main source of A2M expression. The A2M expression signals in secretory endothelium were 2- to 3-fold stronger than the proliferative endothelium, suggesting transcriptional activation of A2M expression in the secretory endothelium. Glandular expression was observed in secretory endometrium from two patients with endometriosis. Ectopic endometrial tissues also produced A2M. A2M at concentrations of 400-500 mumol/L significantly inhibited blastocyst development of mouse embryos in vitro. A2M is expressed predominantly by the endometrial endothelial cells and may be involved in endometrial physiology. Physiological concentrations of A2M inhibit mouse embryo development in vitro, suggesting that endometrial production of A2M may play a role in regulating preimplantation embryo development.

Increased expression of complement component 3 in human ectopic endometrium compared with the matched eutopic endometrium.

OBJECTIVE: To compare the gene expression of complement component 3 (C3) in human eutopic and ectopic endometrium. DESIGN: A prospective, controlled study. SETTING: Academic hospital. PATIENT(S): Women with documented endometriosis. INTERVENTION(S): Eutopic and ectopic endometrial tissues were collected simultaneously at laparoscopy. MAIN OUTCOME MEASURE(S): Detection of C3 messenger RNA (mRNA) by in situ hybridization and C3 protein by immunohistochemistry and Western blot. RESULT(S): Expression of C3 mRNA increased in ectopic endometrium compared with that in the matched eutopic endometrium. The quantitative analysis of C3 mRNA by grain count (mean +/- SE) showed 175.60 +/- 40.02 and 39.97 +/- 8.17 grains per micron2 in ectopic and eutopic glands, respectively, and 67.65 +/- 29.82 and 15.02 +/- 5.80 grains per micron2 in ectopic and eutopic stroma, respectively. Expression of C3 mRNA in ectopic glands was significantly higher than that in eutopic glands. The pattern of immunoreactive staining of C3 protein was consistent with that of C3 mRNA. A higher level of C3 protein in ectopic endometrium than eutopic endometrium was detected by immunohistochemistry and Western blot. CONCLUSION(S): Expression of C3 mRNA and protein significantly increased in human ectopic endometrium compared with that in the matched eutopic endometrium.

Localization of the expression of complement component 3 in the human endometrium by in situ hybridization.

C3 production by the human endometrium has been previously described. The objective of the current study was to localize the site of expression and regulation of the third component of complement, C3, in the endometrium. Eight secretory and eight proliferative archival endometrial samples from hysterectomy and endometrial biopsy specimens were used for in situ hybridization analysis. This analysis was performed with a radiolabeled riboprobe synthesized from a 736-bp template representing sequence 1944-2680 of the human C3 complementary DNA. Duplicate sections were hybridized with sense and antisense riboprobes. Resultant autoradiograms were analyzed qualitatively by light- and darkfield microscopy. In proliferative endometrium, minimal expression of C3 was observed and was limited to a few stromal patches and glands throughout the section. In the secretory samples, prominent C3 expression was observed in both the glands and stroma of the basalis layer. Endometrial lymphocytes did not express C3. Endometrial stromal and glandular cells express the C3 gene. Endometrial lymphocytes did not express C3, but other nondistinct lymphoid elements scattered in the stroma may be expressing C3. There was a visibly more intense expression of C3 in the basalis layer of the secretory endometrium than in proliferative endometrium. The spatial and temporal pattern of C3 expression may have implications in normal menstrual physiology and in the immunological response of the endometrium to the invading trophoblast during placentation.

Immunohistochemical localization of alpha-2 macroglobulin receptor/low-density lipoprotein receptor-related protein, receptor-associated protein, and Gp330 in the human endometrium.

OBJECTIVE: We investigated the expression of alpha-2 macroglobulin receptor/low-density lipoprotein receptor-related protein (LRP), an analogous receptor Gp330, and the 39-kd receptor-associated protein (RAP) in the human endometrium. METHODS: Specific polyclonal and monoclonal antibodies against LRP, Gp330, and RAP were used in standard immunohistochemical analyses of normal secretory and proliferative archival endometrial tissue. RESULTS: There was prominent and uniform stromal staining for LRP in secretory and proliferative endometrium. In both phases, 10-25% of glands stained positive for Gp330, with no appreciable stromal staining for Gp330. Also in both phases, 15-30% of glands stained positive for RAP, with apical staining in proliferative glands and uniform staining in secretory glands. Stromal staining for RAP was patchy and appeared to be more intense in the secretory samples than in the proliferative ones. CONCLUSIONS: The human endometrium expresses LRP, RAP, and Gp330. These receptor proteins are known to be involved in endocytosis of multiple ligands. Some of these ligands, such as proteases, plasminogen activators, and cytokines, are produced by the endometrium and play a role in endometrial remodeling and receptivity to implantation. By virtue of their in vitro properties, it is conceivable that endometrial LRP, Gp330, and RAP are involved in endometrial physiology.

Perinatal outcome of singleton pregnancies conceived by in vitro fertilization: a controlled study.

To determine whether conception by in vitro fertilization and embryo transfer (IVF) predisposes to perinatal complications, the obstetric records of 54 women delivered of singleton pregnancies after conception by IVF were examined. Control women were matched for age, parity, race, year of delivery, diethylstilbestrol exposure and medical problems; another group of women who conceived after infertility treatment was matched in similar fashion. IVF patients showed a longer first stage of labor than previously infertile women, experienced a greater intrapartum blood loss than control or previously infertile women, and showed a trend toward a higher cesarean delivery rate than control women. The differences noted probably do not arise from the physiology of IVF, and although some differences are statistically significant, they are of minimal clinical significance. Singleton pregnancies arising after IVF should not be considered as high risk in the absence of other predisposing factors.

Delivery of the low birthweight and the very low birthweight breech: cesarean section or vaginal delivery?

We studied neonatal survival rates, APGAR scores, and length of hospital stay in 199 singleton breeches weighing 1000-2500 grams at birth. We found that in the birthweight range of 1000-1750 grams, breeches who were delivered by cesarean section had a significantly higher survival rate (74%) than those who were delivered vaginally (36%, p less than 0.01), however, in the birthweight range of 1751-2500 grams, there was no significant difference in the survival rates between breeches delivered abdominally and those delivered vaginally. The 1-minute and the 5-minute APGAR scores and the length of the hospital stay were not significantly different between the abdominal and the vaginal delivery groups in either birthweight range. The data indicate that the very low birthweight breech (less than or equal to 1750 grams) may benefit from a prophylactic cesarean section.