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This investigation examined the hypothesis that release of K+accounts for EDHF activity by comparing relaxant responses produced by ACh and KCl in human subcutaneous resistance arteries.Resistance arteries (internal diameter 244±12 μm, n=48) from human subcutaneous fat biopsies were suspended in a wire myograph. Cumulative concentration‐response curves were obtained for ACh (10−9– 3×10−5M) and KCl (2.5 – 25 mM) following contraction with noradrenaline (NA; 0.1 – 3 μM).ACh (Emax99.07±9.61%; −LogIC507.03±0.22; n=9) and KCl (Emax74.14±5.61%; −LogIC502.12±0.07; n=10)‐induced relaxations were attenuated (P<0.0001) by removal of the endothelium (Emax8.21±5.39% and 11.56±8.49%, respectively; n=6 – 7).Indomethacin (10 μM) did not alter ACh‐induced relaxation whereas L‐NOARG (100 μM) reduced this response (Emax61.7±3.4%, P<0.0001; n=6). The combination of ChTx (50 nM) and apamin (30 nM) attenuated the L‐NOARG‐insensitive component of ACh‐induced relaxation (Emax: 15.2±10.5%, P<0.002, n=6) although these arteries retained the ability to relax in response to 100 μMSIN‐1 (Emax127.6±13.0%, n=3). Exposure to BaCl2(30 μM) and Ouabain (1 mM) did not attenuate the L‐NOARG resistant component of ACh‐mediated relaxation (Emax, 76.09±8.92, P=0.16; n=5).KCl‐mediated relaxation was unaffected by L‐NOARG+indomethacin (Emax; 68.1±5.6%, P=0.33; n=5) or the combination of L‐NOARG/indomethacin/ChTx/apamin (Emax; 86.61±14.02%, P=0.35; n=6). In contrast, the combination of L‐NOARG, indomethacin, ouabain and BaCl2abolished this response (Emax, 5.67±2.59%, P<0.0001, n=6).The characteristics of KCl‐mediated relaxation differed from those of the nitric oxide/prostaglandin‐independent component of the response to ACh, and were endothelium‐dependent, indicating that K+does not act as an EDHF in human subcutaneous resistance arteries.

Summary: An immunocytochemical double horseradish peroxidase-anti-horseradish peroxidase (PAP) technique has been developed for localising estrogen, testosterone, and progesterone binding sites in normal ovaries and in epithelial ovarian neoplasms. Estrogen binding sites were present in 45 of normal ovaries, in 45 of benign epithelial neoplasms, and in 58.5 of ovarian adenocarcinomas. The equivalent figures for progesterone binding sites were 49, 65, and 45.2, whilst those for testosterone binding sites were 43, 40, and 60.5. Steroid binding was related neither to the grade of malignancy in epithelial neoplasms nor to the presence of metastases in cases of ovarian adenocarcinomas. The simultaneous presence of both estrogen and progesterone binding sites or of both estrogen and testosterone binding sites in ovarian adenocarcinomas was, however, associated with good differentiation. Evidence is presented to suggest that the binding sites demonstrated were specific, and it is suggested that the immunohistochemical demonstration of sex steroid hormone binding capacities in ovarian adenocarcinomas may be of value as a predictive marker for response to hormonal therapy

A study of the immunoglobulin content of endometria from women with and without endometriosis has shown that, in women with endometriosis, both 1gG and IgA are more commonly found in the interstitium of the endo-metrial stroma than is the case in endometria from women without this disease. It is thought that the increased stromal content of immunoglobulins in endometriosis is simply a passive reflection of elevated serum IgG and IgA levels. Both the incidence and extent of positive endometrial glandular epithelial staining for IgG and IgA are markedly increased in women with endometriosis: the excess of intraepithelial IgA is probably simply a consequence of the excess of stromal IgA, but the increased epithelial staining for IgG lends support to the concept that many women with endometriosis develop autoantibodies directed against an endometrial epithelial antigen. No relationship could be demonstrated, however, between IgG deposition in the endometrium of women with endometriosis and a history of infertility.

Secretory differentiation of endometrium after multiple follicular stimulation using gonadotrophin releasing hormone and human menopausal gonadotrophin has been studied both histologically and immunohistochemically in 30 women undergoing in-vitro fertilization treatment. None had embryo transfer. Patients were randomly allocated to receive luteal phase support with a single dose of human chorionic gonadotrophin. The latter failed to produce any significant enhancement of endometrial structure or secretions. Appropriate glandular morphology was present in a greater proportion of those who were successfully stimulated than those who responded poorly. However, defective secretion of the cycle-dependent component studied, using monoclonal antibody D9B1, was demonstrated in two-thirds of cases regardless of the ovarian response. Early vascular maturation in the stroma was a common finding, and was thus considered as a feature of structural modulation of these endometria.

The expression of an oestrogen receptor-related antigen has been measured by microspectrophotometry in the glandular epithelium and stroma of tissue sections from endometrial biopsies of women administered exogenous replacement of sex steroids because of premature ovarian failure. We have demonstrated that previously dormant endometrium is capable of expressing an antigen related to oestrogen receptors and that its distribution is influenced by the sex steroid environment. Immunohistochemical staining was more intense (P less than 0.001) in the glandular compared to the stromal cells in tissue exposed solely to oestradiol. With the administration of progesterone, the stromal staining intensity increased (P less than 0.0001) and that of the glandular staining decreased (P less than 0.0001). No assumption, however, has been made to infer that colour is stoichiometrically related to the concentration of antigen present. Further validation of immunohistochemical procedures is required because histological localization and measurement of immunohistochemical staining may be straightforward, but its relationship to antigen concentration remains unproven.

Subinvolution of Uteroplacental arteries is a well-recognized cause of hemorrhage in the postpartum period. Although the physiological changes in these arteries during pregnancy are well documented, the sequence of events in normal involution is largely unknown. A recent immunohistochemical study has raised the possibility of an abnormal interaction between maternal uterine cells and fetal trophoblast in Subinvolution. An indirect immunoperoxidase technique was used to compare deposition of complement components and immunoglobulin in subinvoluted and normally involuted Uteroplacental arteries in 25 cases of postpartum hemorrhage. Deposits of C1q, C3d, C4, and C9 were detected within the walls of normally involuted vessels, whereas deposition of Clq, C3d, and C4 was absent in subinvoluted vessels; C9 was detected only focally. Deposition of immunoglobulins G, A, and M mirrored those described for complement components. The results suggest that immunological factors are necessary for the process of normal involution of Uteroplacental arteries and are deficient in subinvoluted vessels.

An immunohistochemical study was made of cathepsin-D protein expression in each of the three main types of uterine cervical carcinoma (squamous carcinoma, adenosquamous carcinoma and adenocarcinoma) with particular reference to lymph node status and prognosis. Of the 61 cases, 54.1% showed cytoplasmic staining in more than 2.5% of tumour cells counted. Cathepsin-D expression was significantly higher in adenocarcinoma (mean -3.128) than in squamous carcinoma and adenosquamous carcinoma (mean -3.709,P=0.047 using logit transformation). Cathepsin-D had no prognostic value in any of the three tumour types. No relationship was found between cathepsin-D staining and lymph node status and there was no advantage in adding cathepsin-D values to lymph node status. These results suggest that immunostaining for cathepsin-D protein expression is unlikely to be of use as a prognostic marker.

Summary The filamentous components of the cytoskeleton in smooth muscle cells of leiomyomata and normal myometrium were studied by immunohistochemistry and electron microscopy. Fourteen patients hysterectomised for non-malignant disease provided leiomyomata of conventional histological type and histologically normal myometrium: four samples of fetal myometrium were studied by immunohistochemistry alone. All samples of leiomyoma and myometrium were strongly positive for a-smooth muscle actin and desmin, the latter often as paranuclear columns or granules. Vimentin was also stained in most samples but less intensely, while cytokeratin stained in about half the samples with an intensity comparable to that of vimentin. By electron microscopy, myofilaments with focal densities were abundant in both normal myometrium and leiomyomata. Intermediate filaments corresponding to the desmin and vimentin demonstrated by immunohistochemistry were also recognised in a variety of architectural arrangements. At one extreme, comparatively small numbers of filaments were loosely distributed around membranous organelles; at the other, filaments formed conspicuous aggregates, largely excluding other organelles and corresponding to the paranuclear granules seen by immunohistochemistry. A comparison of these findings with those of the literature and comments on the possible significance and origin of these aggregates are provided.

The purpose of this study was to assess the ability of the anti-progestin onapristone administered in the immediate post-ovulatory period to disrupt endometrial differentiation as a potential method of fertility control. In all, 10 healthy female volunteers were given 400 mg onapristone 2 days after the mid-cycle luteinizing hormone (LH) surge in urine (LH+2). An endometrial biopsy was taken 4 or 6 days after the LH surge (i.e. LH+4 or LH+6) in a control cycle and on the corresponding day of the treatment cycle. Biopsies were assessed for histological dating and immunolocalization of oestrogen receptors, progesterone receptors and 15-hydroxyprostaglandin dehydrogenase (PGDH). On day LH+12, blood was taken for the measurement of insulin-like growth factor binding protein-1 (IGFBP-1) and placental protein 14 (PP14). Hormonal measurements in blood and urine were used to monitor the effects on the menstrual cycle. In addition, the concentration of cortisol in plasma was measured to determine if this dose of onapristone exerted significant anti-glucocorticoid activity. Treatment with onapristone retarded the development of secretory changes within the endometrium without affecting the length of the luteal phase. Intense nuclear immunostaining of oestrogen and progesterone receptors was evident in glands and stroma after treatment, suggesting that the progesterone-dependent down-regulation of steroid receptors was inhibited by the anti-progestin. Onapristone also affected the production of luteal phase endometrial proteins, as judged by the pronounced reduction in immunostaining of PGDH within the glands and the significant reduction in plasma concentrations of PP14. However, plasma concentrations of IGFBP-1 did not differ between cycles. Onapristone did not appear to exert significant anti-glucocorticoid activity because concentrations of cortisol were unaffected. These findings suggest that onapristone could potentially be used as a method of postovulatory fertility control.

Following an ovulatory control cycle, six women took 2 mg of mifepristone daily for 30 days. Endometrial biopsies were collected in the control cycle between 7 and 11 days after the plasma luteinizing hormone (LH) surge and on the corresponding day of the treatment cycle (days 19-28). In order to investigate the effects of unopposed oestrogen on the endometrium, persistent proliferative endometrium was obtained from six women with anovulatory infertility due to polycystic ovarian syndrome (PCOS) on a similar cycle day (days 21-23) following a progestogen-induced withdrawal bleed. Endometrium was evaluated for histology and immunolocalization of oestrogen receptors (ER), progesterone receptors (PR) and the cell proliferation markers [proliferating cell nuclear antigen (PCNA) and Ki67]. Treatment with mifepristone inhibited ovulation in four of the six subjects. In the two subjects in whom ovulation did occur, secretory transformation was delayed, suggesting that successful implantation of a blastocyst would be unlikely. In subjects who remained anovulatory during treatment, the histology and pattern of steroid receptor expression was similar to proliferative phase endometrium. In women with PCOS, mitoses and intense immunostaining for ER, PR and cell proliferation markers were observed in both glands and stroma. Although PCNA and Ki67 immunostaining were also present in mifepristone-treated endometrium from subjects who did not ovulate, there were no mitoses and significantly less ER immunostaining in spite of exposure to unopposed oestrogen for a similar duration. Since PCNA and Ki67 detect cells throughout all stages of the cell cycle this would suggest that mifepristone might affect the entry of cells into the mitotic phase of the cell cycle and, therefore, might prevent endometrial hyperplasia. These findings add further evidence to support the contraceptive potential and antiproliferative activity of daily low dose mifepristone.

Irregular bleeding remains a common reason for the discontinuation of progestin-only contraception. The levonorgestrel releasing intrauterine system (LNG-IUS) has profound morphological effects upon the endometrium. Specific features are gland atrophy and extensive decidual transformation of the stroma. Morphological changes in the endometrium may be associated with perturbation of mechanisms regulating normal endometrial function. This study describes endometrial stromal and glandular features prior to and up to 12 months following insertion of the LNG-IUS. Comparison is made with first trimester decidua. In order to elucidate further mechanisms governing endometrial function with local intrauterine delivery of LNG, we here report histological features consistent with decidualization; a significant increase in granulocyte-macrophage colony stimulating factor (GM-CSF) immunoreactivity in decidualized stromal cells; glandular and stromal prolactin receptor expression and an infiltrate of CD56 + large granular lymphocytes and CD68 + macrophages. We are unaware of previous reports which have documented longitudinally both morphological and functional observations in endometrium exposed to local intrauterine levonorgestrel delivery. These studies demonstrate that long-term administration of intrauterine levonorgestrel results in features of altered morphology and function. No correlation was apparent between the end points in the study and the bleeding patterns described by the subjects. Further evaluation of these features in the context of menstrual bleeding experience may contribute to a better understanding of this troublesome side-effect which often leads to dissatisfaction and discontinuation of the intrauterine system.

An immunoperoxidase technique has been utilised for the demonstration of follicle stimulating hormone (FSH), luteinising hormone (LH) and prolactin (PRL) binding sites in normal human ovaries and in a wide range of benign and malignant epithelial tumours of the ovary. The incidence of FSH, LH and PRL binding was, respectively, 32%, 41% and 39% in normal ovaries, 30%, 18.5% and 22.5% in benign epithelial tumours and 51%, 32% and 43% in malignant epithelial neoplasms. The incidence of FSH binding was significantly higher in malignant epithelial neoplasms than in either normal ovaries or benign epithelial tumours but otherwise no correlation was found between hormone binding capacity and the degree of malignancy of epithelial ovarian tumours, the histological type of the tumour, the degree of differentiation of the malignant epithelial tumours or the presence or absence of metastatic disease. Well differentiated malignant tumours did, however, tend to stain more strongly than did poorly differentiated neoplasms, thus suggesting that the number of binding sites per cell tends to decrease with decreasing degrees of differentiation.

Tumour cords have been measured in 33 cases of squamous cell carcinoma (SCC) of the bronchus and 19 cases of SCC of the uterine cervix. The overall mean cord radius for SCC in both sites was 104 microns, similar to the overall mean for various tumours in rodents. For tumour cells adjacent to blood vessels in cords of SCC, the mean Mitotic Index was 2.1% and from this value a rapid potential doubling time could be inferred (approximately 31 to 66 h). The proportion of dead cells within cords of cervical SCC was higher than in animal tumours. Using measured values for cord radius and published equations that describe the diffusion and consumption of oxygen in metabolising tissue, an attempt was made to calculate the oxygen partial pressure in vessels of cords of these SCC.

Data are presented on serum levels of the pregnancy-associated placental protein (PP14) in an XO mother, pregnant after frozen embryo transfer. Steroid replacement before and during early pregnancy was provided with transdermal oestradiol and vaginal micronized progesterone. Characterization of a pre-treatment cycle indicated a physiological response by the endometrium. The pregnancy was associated with normal maternal blood levels of all hormones considered responsible for maintenance of early gestation but with consistently subnormal serum levels of PP14. Hitherto, PP14 has been believed to be an endometrial protein arising from the decidua during pregnancy, under the influence of ovarian steroids. The findings in this case suggest that other factors are involved in its production, possibly under the control of, or originating from, the maternal ovary.

FOCAL nodular hyperplasia of the liver is a non-neoplastic lesion (Edmondson, 1956, 1958) and under ordinary circumstances, it is an innocuous condition discovered incidentally at laparotomy, autopsy or on abdominal examination (Knowles and Wolff, 1976). In patients taking steroid contraceptives or in pregnancy, the lesions may grow rapidly, undergo necrosis and rupture (Mays et al., 1974; Knowles and Wolff, 1976; Christopherson and Mays, 1977; Fechner, 1977; Kent et al., 1978; Kinch and Lough, 1978).