Analysis of WT1 in granulosa cell and other sex cord-stromal tumors

The molecular genetic events involved in the etiology of granulosa cell, Sertoli cell, and Leydig cell tumors are unknown. The expression of the Wilms' tumor suppressor gene WT1 in granulosa and Sertoli cells prompted us to analyze this gene for mutations in 11 granulosa cell tumors, three Leydig cell tumors, and one Sertoli/Leydig cell tumor. Although most of these tumors express WT1 mRNA, none harbors a WT1 mutation in the zinc finger domains where > 90% of WT1 mutations in sporadic Wilms' tumors have been found. In addition we were able to exclude tumor-specific loss of heterozygosity in 13 of 15 cases. Taken together these results suggest that the WT1 gene is unlikely to play an important role in the development of sex cord-stromal tumors.     

Use of a long-acting gonadotropin-releasing hormone agonist for treatment of steroid cell tumors of the ovary

OBJECTIVE: To report a complete serologic response in a 50-year-old women who received long-acting gonadotropin-releasing hormone agonist (GnRH-A) therapy for steroid cell tumor of the ovary, not otherwise specified. DESIGN: Case report. SETTING: University hospital-based reproductive biology unit. PATIENT(S): A 50-year-old female patient exhibited persistent elevation of T (>2.0 ng/mL) after surgery for steroid cell tumor of the ovary, not otherwise specified, stage IIA for 3 months. This elevation suggested the presence of some residual active tumor. INTERVENTION(S): All tumor evaluations, including those for tumor markers, a thorough physical examination, imaging studies, and evaluations of nuclear medicine studies were negative except for elevated serum T levels. The patient was treated with GnRH-a between the fourth month and sixth month postoperatively. MAIN OUTCOME MEASURE(S): Serum levels of T and tumor survey. RESULT(S): The serum T levels returned to normal limits after administration of the first dose of GnRH-a. Follow-up of tumor survey was negative. The patient was alive and free of disease 26 months after treatment with GnRH-a. CONCLUSION(S): GnRH-a may be an alternative choice as adjuvant therapy for managing a persistent or recurrent hormone-producing steroid cell tumor of the ovary.     

Management of malignant germ cell tumors of the ovary

A high sensitive method for detecting the change of microsomal membrane surface oligosaccharides was developed to study the regulatory role of lipid- or peptide-linked mannoside of endoplasmic reticulum in synaptic functions. The binding of concanavalin A to the microsomal membrane surface was measured quantitatively using a microgram-order of rat brain microsomal proteins. The fluorescence polarization of concanavalin A (Con A)-fluorescein isothiocyanate (FITC) conjugate bound to the membrane was analyzed to quantitate the change of binding constant and the number of binding sites. As a control, the non-specific binding of bovine serum albumin-FITC conjugate was measured by the same technique. We measured the change of fluorescence intensity of membrane-bound FITC conjugates by the flow cytometry and found that the intensity of FITC conjugate bound to the membrane increased more than that of free form of the probe. We observed that the alpha-mannosidase-treatment of rat brain microsomes resulted in the increase of binding constant of Con A to the microsomal surface without significant loss of binding sites.     

Inhibin A is a sensitive and specific marker for testicular sex cord-stromal tumors

We compared the expression of inhibin A, chromogranin, synaptophysin, S-100 protein, cytokeratins AE1/AE3, 7, and 20, and estrogen and progesterone receptors in testicular sex cord-stromal tumors: 11 Sertoli cell tumors, 3 Sertoli cell adenomas (nodules), 26 benign Leydig cell tumors, 7 malignant Leydig cell tumors (defined clinically by metastatic behavior), and a variety of germ cell tumors. Inhibin was the most sensitive marker, expressed in 91% of the Sertoli cell tumors and 100% of the Sertoli cell adenomas and Leydig cell tumors. The non-neoplastic Sertoli and Leydig cells invariably stained for inhibin. Conversely, no germ cell tumors were immunoreactive. One testicular tumor of the adrenogenital syndrome was immunoreactive. Neuroendocrine marker immunoreactivity was variable. Chromogranin was expressed in the non-neoplastic Sertoli and Leydig cells, 82% of the Sertoli cell tumors, 92% of the benign Leydig cell tumors, and 43% of the malignant Leydig cell tumors. Synaptophysin was expressed in the non-neoplastic Sertoli and Leydig cells, 45% of the Sertoll cell tumors, and 70% of the Leydig cell tumors, in approximately similar proportions between the benign and malignant Leydig cell tumors. S-100 protein was expressed in 64% of the Sertoli cell tumors, 8% of the benign Leydig cell tumors, and none of the malignant Leydig cell tumors. Cytokeratins AE1/AE3 were expressed in 64% of the Sertoli cell tumors and 42% of the Leydig cell tumors, with similar proportions in the benign and malignant cases. Estrogen and progesterone receptor expression were identified in 24 and 39% of benign and malignant Leydig cell tumors, respectively. We conclude that inhibin is a characteristic marker for Sertoli and Leydig cells and that it serves to differentiate testicular sex cord-stromal tumors from germ cell tumors.     

Immunohistochemistry of unclassified sex cord-stromal tumors of the testis with a predominance of spindle cells

Onpi-to (TJ-8117) is a herbal medicine composed of five crude drugs (Rhei Rhizoma, Glycyrrhizae Radix, Ginseng Radix, Zingiberis Rhizoma and Aconiti Tuber). Our previous experiments have demonstrated that TJ-8117 suppressed the development of glomerulosclerosis and retarded the deterioration of renal function in 5/6 nephrectomised rats. In the present study, the effects of TJ-8117 and (-)Epicatechin-3-O-gallate (ECG), which is a component of Rhei Rhizoma, on glomerular cell proliferation, extracellular matrix accumulation and glomerular hypertrophy were investigated in 5/6 nephrectomized rats. Male Wistar rats (170-180 g) were subjected to 5/6 nephrectomy, and TJ-8117 (0.32%, 0.64%) or angiotensin-converting enzyme inhibitor, captopril (CAP 0.08%) was administered daily by mixing in normal chow and ECG (2 mg, 8 mg/100 ml) by drinking water from the day after 5/6 nephrectomy. Following 5/6 nephrectomy, glomerular cell poliferation was increased and reached a maximum at 1 week in the untreated control rats, but was suppressed significantly at 1 and 2 weeks after treatment with TJ-8117 and at only 1 week after treatment with CAP. Extracellular matrix accumulation was detected after 1 week and increased gradually until 4 weeks in the control rats, whereas it was significantly inhibited in both the TJ-8117- and CAP-treated rats. In addition, immunohistochemistry revealed that TJ-8117 significantly inhibited the increase of fibronectin, and tended to reduce type I and type IV collagen at 4 weeks. Furthermore, TJ-8117 suppressed glomerular hypertrophy at 4 weeks. Systolic blood pressure (SBP) and urinary protein excretion (UP) were higher in the control rats than sham-operated rats. TJ-8117 prevented this increase of SBP and UP at 1 week. ECG also suppressed glomerular cell proliferation and the increase of SBP and UP at 1 week after 5/6 nephrectomy. These findings suggest that ECG was one of active components of TJ-8117. These results suggest that TJ-8117 suppressed proliferating changes in glomeruli at an early stage in 5/6 nephrectomized rats, and inhibited the development of glomerulosclerosis.     

Sex and androgenic steroid receptor expression in hepatic adenomas

Sex hormones and anabolic-androgenic steroids are implicated in the development and progression of hepatic adenomas (HA). We studied the expression of their receptors in HA and adjacent liver. Archival tissue sections of 27 HA (16 resections, four needle biopsies, seven aspirations) from 18 patients, and the adjacent liver, were immunostained with monoclonal antibody to estrogen receptor (ER, 1/80) (Dako, Carpinteria, CA), progesterone receptor (PR, 1/50) (BioGenex, San Ramon, CA), and androgen receptor (AR, 1/80) (BioGenex). An avidin-biotin complex technique was used with microwave antigen retrieval. Nuclear expression was assessed as 1+ to 3+ intensity, with semiquantitation of the percentage of nuclei immunopositive. Five percent or more nuclei immunopositive was regarded as positive. The 18 patients included 16 females of 34 years mean age (range, 16 to 49) with an available history of oral contraceptives in five; the two men were 24 and 30 years, with no history of androgenic steroids. ER, PR, and AR were present in seven (26%) (1+/-2+ intensity, 5% to 10% of nuclei) of HA, seven (26%) (1+/-2+ intensity, 5% to 30% of nuclei) and nine (33%) (1+/-3+ intensity, 5% to 80% of nuclei), respectively. In the adjacent liver in 11 cases, there were one (9%) ER, (2+ intensity, 5% of nuclei), four (36%) PR (1+/-2+ intensity, 5% to 20% of nuclei), and two (18%) AR (2+/-3+ intensity, 10% of nuclei). Receptors are present and may mediate the action of sex hormones or androgenic steroids on HA and adjacent liver, but in less than one third of patients. This may have therapeutic implications.     

Mural nodules in common epithelial tumors of the ovary

The occurrence of mural nodules in cystic common epithelial tumors of the ovary is well established, but differences in terminology and difficulties in histopathologic interpretation have hampered adequate understanding of their differential diagnoses and prognoses. Using immunohistochemistry and ultrastructural analyses supplementally, we studied two cases, one a serous cystadenocarcinoma with nodules of undifferentiated sarcoma, the other a mucinous cystadenocarcinoma with mural nodules of anaplastic carcinoma. Based on these cases and a review of 48 cases in the literature, we propose a standardized terminology. Mural nodules may be either reactive or neoplastic. Neoplastic mural nodules may be composed of benign elements, carcinoma, carcinoma with reactive elements, sarcoma, or an admixture of carcinoma and sarcoma; the latter category has some similarities to malignant mixed mesodermal tumors. The prognosis of patients with malignant mural nodules is poor, with 50% mortality. Strict morphologic criteria supplemented by immunohistochemistry aids in the sometimes difficult differential diagnosis among these types of mural nodules.     

Sex steroid hormones and genetic susceptibility to breast and prostate cancer

In SPECT, both the noise affecting the data and the discretization of the inverse Radon transform are responsible for the ill-posed nature of the reconstruction. To constrain the problem, we propose a regularized backprojection method (RBP) which takes advantage of the relationships existing between the continuity properties of the projections and those of the reconstructed object. The RBP method involves two stages: first, a statistical model (the fixed-effect model) is used to estimate the noise-free part of the projections. Then, the filtered projections are reconstructed using a backprojection algorithm (spline filtered backprojection) which ensures that the reconstructed object belongs to a space consistent with that containing the projections. The method is illustrated using analytical simulations, and the RBP approach is compared to the conventional filtered backprojection. The effect on the reconstructed slices of the parameters involved in RBP is studied in terms of spatial resolution, homogeneity in uniform regions and quantification. It is shown that appropriate combinations of these parameters yield a better compromise between homogeneity and spatial resolution than conventional FBP, with similar quantification performances.     

Value of CT scan and MRI in primary tumors of the ovary

Characterisation of an ovarian mass is of utmost importance in the preoperative evaluation of an ovarian neoplasm. It enables the surgeon to anticipate carcinoma of the ovary before the operation, so that adequate procedures are planned. Although, ultrasonography (US) remains the foremost imaging modality for screening patients with adnexal lesions, computed tomography (CT) recently has proved to be of value in the characterisation and management of tumors of the ovary. Magnetic resonance (MR) imaging has also been shown to have a high degree of diagnostic specificity for certain types of ovarian masses, such as dermoid cysts, and endometriomas. However, the potential of MR imaging to characterize tumors of the ovary has not yet been established. This study assesses the value of MR imaging and CT for the purpose of predicting the malignancy of ovarian lesions, and comparing findings of MR imaging with those of CT.     

Sex steroid metabolism and menstrual irregularities in the exercising female. A review

This article aims to clarify why, and by which mechanisms, exercise may influence the normal menstrual cycle. Therefore, the vast amount of literature on this subject is reviewed and a critical appraisal of the most widespread hypotheses if offered. The strikingly low body mass which frequently accompanies exercise-related menstrual irregularities (ERMI) has led some authors to develop a hypothesis which postulates that a critical percentage of body fat is essential to trigger normal menstruation. The relevance of any reference to anorexia nervosa to support this view lacks consistency: female athletes differ in many ways from patients with anorexia nervosa, not least in their excellent physical status which is essential to deliver first-class performances. ERMI is not identical to the so-called female athlete triad, a complicated pathology that involves ERMI, premature osteoporosis and disordered eating. ERMI itself does not seem to have any substantial pathological effects as long as attention is paid to preventing osteoporosis or stress fractures which may result from prolonged hypo-estrogenaemia. In the female athlete with ERMI who wishes to conceive, the accompanying subfertility may necessitate a response other than a prompt reduction in training intensity, as this is hardly a first choice for any top athlete. During recent years, a number of prospective studies have greatly contributed to our understanding of the complexity of the mechanisms involved in ERMI. Older hypotheses, such as those considering hyperprolactinaemia as the cornerstone of ERMI, have now been firmly rejected. The present hypotheses emphasise the importance of caloric deficiency and limited energy availability, although they still fail to identify the actual mechanism that causes ERMI. There is, however, evidence that ERMI is produced by a disturbance of the hypothalamic gonadotrophin-releasing hormone oscillator. This disturbance is caused by either an insufficient estrogen or progesterone feedback or by an imbalance of local opioid peptide and catecholamine activities mediated by gamma-aminobutyric acid (GABA), corticotrophin-releasing hormone and insulin-like growth factor-1. More recent experiments have also linked ERMI with changes in steroid metabolism, in particular, an increasing activity of catecholestrogens possibly leading to enhanced intracerebral noradrenaline (norepinephrine) levels that may interfere with normal gonadotrophin release. This article demonstrates that the outcome of the many studies of ERMI is characterised by much controversy and numerous methodological flaws. The importance and complexity of some recent findings necessitate a comprehensive study which links older and newer findings within a critical perspective.     

Asymmetric effects of acute hemiovariectomy on steroid hormone secretion by the in situ ovary

The acute effects of hemiovariectomy on progesterone, testosterone, estradiol, and luteinizing hormone (LH) concentrations in serum were studied in rats under the following experimental conditions: control, shamoperated (left or right), hemiovariectomized, bilateral adrenalectomized, and hemiovariectomized plus bilateral adrenalectomized. One-hour after surgery, the concentration of progesterone and testosterone in the serum of right-side sham-operated rats was significantly higher than in control animals. Testosterone concentration in serum in rats with the right ovary in situ was higher than in sham-operated animals; injecting atropine sulfate 1 h before surgery blocked such increase, while the same treatment to rats with the left ovary remaining in situ resulted in a significant increase of testosterone concentration. Adrenalectomy resulted in an increase of testosterone concentration, which was higher when atropine sulfate was injected before surgery. Our results support the idea that left and right ovaries play different roles in the regulation of hormone secretion, and that such differences are related to ovarian innervation.     

Management of germ cell tumours of the ovary

Malignant ovarian germ cell tumours (OGCT) comprise only 2–5% of all ovarian cancers but are significantly different to epithelial ovarian cancers. They affect women of child bearing age and are much more curable than their epithelial counterparts. In addition, the majority of patients will retain their fertility after multimodal treatment. The small numbers of patients mean that randomised controlled trials of chemotherapy, the gold standard test of treatment effectiveness in other malignancies, have proved impossible to perform. The different types of OGCT have variable degrees of chemosensitivity and differing prognoses. Treatment outcomes are also dependent on the stage of disease at diagnosis. In this article, dysgerminomas and non-dysgerminomas are analyzed separately, as there are notable differences in their behaviour and outcomes. It is difficult to think of many diseases in which prognosis has improved as greatly as ovarian germ cell tumours and this is due to modern combination chemotherapy. Like the treatment of testicular cancer, this represents one of the successes of modern medicine.     

P53 protein expression and DNA ploidy in common epithelial tumors of the ovary

OBJECTIVE: To investigate p53 protein expression and DNA content in imprints from surgical biopsies of common epithelial tumors of the ovary. STUDY DESIGN: The study was based on 60 cases of epithelial tumors of the ovary (15 benign, 3 border-line and 42 malignant). For the demonstration of p53 protein, immunocytochemical staining with the avidin-extravidin technique was performed using monoclonal antibody p53 DO-7. DNA content was measured by image cytometry after Feulgen staining. RESULTS: There was a strong correlation between p53 expression and aneuploidy, with the difference between diploid and aneuploid tumors statistically significant (P < .001). A correlation was found between DNA ploidy, histologic grade and clinical stage (P < .001 and P < .05), respectively. There was no correlation between DNA ploidy and histologic type (P = .89). No correlation was observed between p53 protein expression and grade or clinical stage of the tumors. Nevertheless, a correlation of p53 expression between early (I, II) and advanced stages (III, IV) (P < .05) was observed. All benign and borderline tumors were diploid and did not express p53 protein. CONCLUSION: The results of the present study and the data in the literature stress the value of p53 expression and DNA ploidy in assessing the malignant potential of common epithelial ovarian cancers. However, the clinical application of these data requires further study.     

Tumor cell procoagulant and urokinase expression in carcinoma of the ovary

BACKGROUND: An association between cancer and increased blood coagulation has been observed for many years. Generally, there is an equilibrium between the coagulation system (fibrin deposition) and the fibrinolytic system (degradation of fibrin by enzymes). However, in malignant disease such as ovarian carcinoma, this equilibrium is disrupted, resulting in the abnormal activation of coagulation or hypercoagulability. Also, evidence indicates that various components of these pathways may contribute to the disorderly characteristics of malignancy, such as proliferation, invasion, and metastasis. PURPOSE: Our purpose was to define the mode of interaction of tumor cells in ovarian carcinoma with both the coagulation (procoagulant-initiated) and fibrinolysis (urokinase-type plasminogen activator-initiated) (u-PA) pathways. METHODS: Studies were performed on acetone-methylbenzoate-xylene-fixed tissue prepared from fresh resected primary tumor specimens from 15 patients with cystic epithelial ovarian carcinoma. None of the patients had received prior treatment. Antibodies were tested on control and tumor tissues in concentrations that provided maximum staining intensity with minimum background staining. Laboratory immunohistochemical techniques used purified, monospecific antibodies to detect coagulant antigens. Tests were performed utilizing antibodies to recombinant human tissue factor; factor VII; factor X; factor XIIIA; high-molecular-weight and low-molecular-weight forms of u-PA; tissue-type plasminogen activator; plasminogen; and the plasminogen activator inhibitors 1, 2, and 3. Monoclonal antibodies used for specific antigen detection included 1-8C6 (fibrinogen), T2G1 (fibrin), and EBM-11 (macrophage-specific). RESULTS: The ovarian tumor cells expressed urokinase-type plasminogen activator in a pattern that was variable in intensity and distribution. Tumor cell plasminogen was not detected. Tumor cells also expressed tissue factor and coagulation pathway intermediates that resulted in local thrombin generation as evidenced by the conversion of fibrinogen (present in tumor connective tissue) to fibrin that was found to hug the surfaces of tumor nodules and individual tumor cells. Detected fibrin could not be accounted for on the basis of necrosis or a local inflammatory cell infiltrate. CONCLUSIONS: These results are consistent with the existence of a dominant tumor cell-associated procoagulant pathway that leads to thrombin generation and hypercoagulability in carcinoma of the ovary. IMPLICATIONS: In ovarian carcinoma the procoagulant pathway may contribute to tumor progression. Clinical trials of therapeutic drugs capable of limiting local coagulability (anticoagulants, protease inhibitors) are indicated in this tumor type.