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Morphological study of ovarian follicular cysts induced by chorionic gonadotropin (300 U) and insulin (Protafane HM; 2.5 U) in adult rats showed that superovulatory dose of chorionic gonadotropin and moderate hyperinsulinemia induced the development of ovarian cysts on days 3-5 in 100% cases. Dynamic study of ovarian morphology showed that changes were reversible and the cysts regressed within 60 days, which confirmed their functional nature.

Vascular endothelial growth factor (VEGF) serum levels and VEGF and cellular expression were prospectively analyzed in 60 patients (group A consisted of asymptomatic patients or patients presenting mild dysmenorrhea; 30 women comprised group B severe dysmenorrhea and/or chronic pelvic pain and/or dyspareunia) who underwent surgery for cystic ovarian endometriosis to asses whether a correlation exists among VEGF serum levels, VEGF cellular expression, and pelvic pain. No differences were found in VEGF serum levels and VEGF cellular expression between both groups. Therefore, we conclude that pain symptoms in ovarian endometriosis are not correlated with VEGF serum levels and VEGF cellular expression.

This study investigated cortisol inactivation by 11beta-hydroxysteroid dehydrogenase (11beta HSD) enzymes in porcine granulosa cells from antral follicles at different developmental stages and in ovarian cysts. In granulosa cells, cortisol oxidation increased threefold with antral follicle diameter (P < 0.001). This trend was paralleled by a threefold increase in NADP(+)-dependent 11beta-dehydrogenase activity in granulosa cell homogenates with follicle diameter. Intact granulosa cells from ovarian cysts exhibited significantly lower enzyme activities than cells from large antral follicles. Neither intact cells norcell homogenates displayed net 11-ketosteroid reductase activities. Since porcine follicular fluid (FF) from large antral follicles and ovarian cysts contains hydrophobic inhibitors of glucocorticoid metabolism by type 1 11beta HSD, this studyalso investigated whether levels of 11beta HSD inhibitors changed during follicle growth and could affect cortisol metabolism in granulosa cells. The extent of inhibition of 11beta HSD1 activity in rat kidney homogenates decreased progressively from 50 +/- 8% inhibition by FF from small antral follicles (P < 0.001) to 23 +/- 6% by large antral FF (P < 0.05). Cyst fluid inhibited 11beta HSD1 activity by 59 +/- 4% (P < 0.001). Likewise, net cortisol oxidation in granulosa cells was significantly decreased by large antral FF (35-48% inhibition, P < 0.05) and cyst fluid (45-75% inhibition, P < 0.01). We conclude that inactivation of cortisol by 11beta HSD enzymes in porcine granulosa cells increases with follicle development but is significantly decreased in ovarian cysts. Moreover, changes in ovarian cortisol metabolism are accompanied by corresponding changes in the levels of paracrine inhibitors of 11beta HSD1 within growing ovarian follicles and cysts, implicating cortisol in follicle growth and cyst development.

The distribution of intermediate filaments (vimentin, cytokeratins, desmin) and microfilaments (alpha-smooth muscle actin and muscle specific actin) was studied immunohistochemically in bovine ovaries, with and without cystic ovarian disease. The immunohistochemically stained area (IHCSA), was quantified by image analysis, to evaluate the expression of these cytoskeletal proteins in the follicular wall of healthy antral, atretic, and cystic follicles. The granulosa cell layer of cystic follicles and atretic follicles had a significantly larger IHCSA for vimentin than did healthy antral follicles. Cytokeratins reacted lightly in the granulosa cells of antral follicles of normal ovaries, whereas granulosa cells of atretic and cystic follicles showed significantly higher IHCSA values. Immunohistochemical localization of desmin, muscle specific actin, and alpha-smooth muscle actin was restricted to the theca externa. This study supports earlier suggestions that strongly positive reactions with vimentin and cytokeratin antibodies observed in the granulosa cells of cystic follicles are due to the reorganization that occurs in the follicle during the process of cystic development, and are associated with changes in the expression of cytoskeletal proteins that are essential to proper cellular functioning.

Background: Female CD-1/Swiss Webster mice subjected to incessant ovulation for 8 months and 12-month breeder mice both developed ovarian inclusion cysts similar to serous cystadenomas. The majority of cysts appeared to be dilated rete ovarii tubules, but high ovulation number resulted in more cortical inclusion cysts. We hypothesized that comparison of inclusion cyst pathology in animals of the same age, but with differences in total lifetime ovulation number, might allow us to determine distinguishing characteristics of the two types of cyst., Methods: Ovaries from breeder mice (BR) or females subjected to incessant ovulation (IO) were compared at 6-, 9- and 12-months of age. Ovaries were serially sectioned and cysts characterized with regard to location and histology, E-cadherin immunoreactivity and rates of BrdU incorporation., Results: Inclusion cysts developed with age in BR and IO ovaries. The majority of cysts were connected to the ovarian hilus. Two cortical inclusion cysts were observed in ten IO ovaries and one in ten BR ovaries. Low or no E-cadherin immuno-staining was seen in the OSE of all mice studied. Conversely, strong membrane immuno-staining was observed in rete ovarii epithelial cells. Variable E-cadherin immunoreactivity was seen in cells of hilar inclusion cysts, with strong staining observed in cuboidal ciliated cells and little or no staining in flat epithelial cells. Two of the three cortical cysts contained papillae, which showed E-cadherin immuno-staining at the edge of cells. However hilar and cortical cysts were not distinguishable by morphology, cell type or E-cadherin immunoreactivity. BrdU incorporation in cyst cells (1.4% [95% CI: 1.0 to 2.1]) was greater than in OSE (0.7% [95% CI: 0.4 to 1.2]) and very few BrdU-labeled cells were observed in rete ovarii at any age. Incessant ovulation significantly increased BrdU incorporation in OSE of older animals., Conclusion: These experiments confirm ovarian inclusion cysts develop with age in the CD-1 mouse strain, irrespective of total ovulation burden. We conclude longer periods of incessant ovulation do not lead to significant changes in inclusion cyst formation or steroidogenesis in CD-1 mice and inclusion cyst type can not be distinguished by morphology, cell proliferation rate or E-cadherin immunoreactivity.

We hypothesized that the special hormonal environment present in animals with cystic ovarian disease (COD) interferes with cellular production of growth factors (GFs). The objective of the present study was to characterize the expression of insulin-like growth factor (IGF)-I, fibroblast growth factor (FGF)-2 and vascular endothelial growth factor (VEGF) in induced COD using immunohistochemistry. We used an experimental model based on the exposure to constant light of adult rats during 15 weeks. We quantified the expression of GFs in cystic and normal ovaries by the Immunohistochemical Stained Area (IHCSA). In animals with COD, a significant reduction in the IHCSA of IGF-I in the follicular fluid, theca and granulosa layers of cysts occurred; and an increase in the interstitial tissue with regard to the control group. We found moderate immunoreactivity of FGF-2 in granulosa and theca layers of secondary and tertiary follicles and lower expression in the granulosa and theca interna layers of cystic follicles. Immunoexpression of VEGF was found in granulosa and theca cells of secondary and tertiary follicles. This study shows changes in the ovarian expression of IGF-I, FGF-2 and VEGF in induced COD. We can propose that an alteration in the control of the follicular dynamic, through the GFs, added to other features, could be involved in the ovarian cyst pathogenesis.

Objective: To investigate the factors related to activation of transforming growth factor-beta 1 (TGF-beta1) at sites of endometriosis., Study Design: TGF-beta1 is activated by plasmin, which is formed when plasminogen is activated by urokinase-type plasminogen activator (uPA). We studied these factors by immunohistochemistry or immunoassay., Results: TGF-beta1 protein was localized mainly in the cytoplasm of glandular epithelial cells in both endometriotic cysts and normal endometrium, but strongly positive immunostaining was significantly more common in cysts. The levels of TGF-beta1, uPA and plasmin/alpha2-plasmin inhibitor complex were all higher in cyst fluid than in peritoneal fluid. There was little uPA protein expression in the glandular epithelium of normal endometrium, but it was prominent in the cytoplasm of glandular epithelial cells from endometriotic cysts, and strongly positive immunostaining was significantly more common in cysts., Conclusion: These results suggest that TGF-beta1 activity is increased at sites of endometriosis due to enhanced production of both uPA and TGF-beta1 by glandular epithelium and because plasmin activates TGF-beta1 after being converted from plasminogen by uPA.

Aim: To determine whether serum CA-125 levels, in addition to tumor size and ultrasonographic findings can help in differentiating benign ovarian cysts from malignant disease., Methods: All postmenopausal women who had undergone explorative laporatomy for a preoperative diagnosis of an adnexal cyst between January 1999 and February 2006 were included if serum CA-125 levels were below 50 IU/ml., Results: Ninety-three patients with ovarian cysts and serum CA-125 levels lower than 50 IU/ml were included. Seventy-five (80%) of the patients (53 unilocular, 22 multilocular) had ovarian cysts < 13 cm. Of 18 patients with ovarian cysts > 13 cm, seven had unilocular and 11 had multilocular cysts. All the patients (n = 77) with a serum CA-125 level < 35 IU/ml had benign histopathology regardless of the tumor size or ultrasonic features. Among 16 patients with CA-125 levels between 35 and 50 IU/ml, two with unilocular cysts > 13 cm and nine with multilocular cysts (3 < 13 cm, 6 > 13 cm) had borderline histopathology., Conclusion: We concluded that when unilocular ovarian cyst size is < 13 cm and serum CA-125 levels are below 35 IU/ml in a postmenopausal woman, the possibility of a benign etiology is most likely.

Endometriosis is an estrogen-correlated benign disease characterized by a marked ability of endometrial-like cells to invade and proliferate outside uterine cavity, resembling for some invasive aspect the cancer growth. The molecular mechanisms regulating endometrial cell invasiveness are mostly unknown, although interactions between extracellular matrix (ECM) proteins and their transmembrane receptors, integrins, are likely to play a central role. In particular, laminin (Ln)-5 could be closely involved, as it is in cancer. We have investigated the expression of Ln-1, Ln-5, and collagen IV (Coll IV) ECM proteins and their receptors, alpha3beta1 and alpha6beta4 integrins, in atrophic, proliferative, and secretive endometrium and in endometriosis. The results show that Ln-5, but not Ln-I and Coll IV, is altered in secretive endometrium as well as in endometriosis tissues. No alterations are observed in atrophic or proliferative endometrium. Consistently, the polarization of both integrin subunits alpha3 and beta1, but not alpha6 and beta4, is altered in secretive endometrium and endometriosis tissues, but not in atrophic and proliferative endometrium. These results seem to suggest that Ln-5 and alpha3beta1 could be involved in the invasive mechanism of endometriosis. The altered expression of Ln-5, by upregulating matrix metalloproteases activity, suggest an invading process similar to that of many cancer processes.

The objective of this study was to evaluate the association between the level of milk production on the day of diagnosis of ovarian cysts and treatment response using the Ovsynch protocol. On the day of cyst diagnosis (Day 0), 260 lactating dairy cows with ovarian cysts were treated with gonadotropin-releasing hormone (GnRH), PGF2alpha on Day 7, GnRH on Day 9, and timed inseminated 16-20 h later (Ovsynch protocol). Pregnancy was determined (by transrectal palpation) between 42 and 49 days after insemination. On Day 0, data for milk production (kg/day), parity, days in milk (DIM), and body condition score (BCS) were recorded. Using the median value for milk production on the day of diagnosis, cows were classified as high producers (>28.5 kg) and low producers (or=0.05). Primiparous cows were more likely (adjusted odds ratio: AOR=3.63; 95% CI: 95% confidence intervals=1.28-10.30; P

Epithelial ovarian cancer (EOC) is often a lethal disease because in many cases early symptoms go undetected. Although research proceeds apace, as yet there are few reliable and specific biomarkers for the early stages of the disease. EOC is an umbrella label for a highly heterogeneous collection of cancers, which includes tumours of low malignant potential, serous cystadenomas, mucinous and clear cell carcinomas, all of which are likely to arise from a number of epithelial cell types and a variety of progenitor lesions. Many, but not all types of EOC are thought to arise from the cells lining ovarian inclusion cysts. In this review, we discuss the hypotheses that have driven our ideas on epithelial ovarian carcinogenesis and examine the morphological and genetic evidence for pathways to EOC. The emergence of laser-capture microdissection and expression profiling by microarray technologies offers the promise of defining these pathways more accurately, as well as providing us with the tools for earlier diagnosis.

Ovarian cancers mostly derive from the monolayer epithelium that covers the ovary. There are currently very few molecular clues to the etiology of this cancer. Bone morphogenetic proteins (BMPs) are required for follicular development and female fertility and are expressed in the ovarian surface epithelium (OSE). We previously reported the expression of human chordin (CHRD), a BMP extracellular regulator, in the ovary. Here we show that CHRD is underexpressed in epithelium ovary cancer and epithelial cancer cell lines as compared with normal tissues and OSE, respectively. Besides, we detected BMP expression in all ovarian cell lines analyzed. To determine the functional relevance of the absence of CHRD mRNA in tumors and cancer cell lines, we studied the effects of CHRD on two cancer cell lines, BG1 and PEO14. Migratory and invasive properties were greatly reduced, whereas cell adhesion to the support was enhanced. In addition, we detected chordin (Chrd) expression in OSE of rat ovaries in a pattern similar to that of BMP4. Altogether, these results suggest that CHRD could participate in regulating BMP activity in normal OSE physiology, and that its mis-expression in OSE may facilitate cancer incidence and/or progression.

Clear cell tumors of the ovary are frequently associated with ovarian endometriosis. Clinicopathologically, it has been suggested that clear cell tumors develop from endometriosis, but there has been little molecular evidence supporting this speculation. Microarray analysis revealed recently that hepatocyte nuclear factor-1beta (HNF-1beta) was significantly upregulated in clear cell carcinoma of the ovary. In the present study, we examined 30 clear cell tumors (26 malignant, three borderline, and one benign) and 40 endometriotic cysts to clarify if differentiation into the clear cell lineage already begins in ovarian endometriosis. All of the 30 clear cell tumors, including borderline and benign ones, showed immunohistochemical expression of HNF-1beta in the nucleus, while other types of ovarian epithelial tumors (endometrioid, serous, mucinous, and Brenner tumors) rarely expressed it. Among 30 clear cell tumors, 17 (56%) cases were associated with endometriosis, and endometriotic epithelium was identified in 12 cases. In nine of the 12 cases, distinct nuclear immunostaining for HNF-1beta was detected in the endometriotic epithelium, as well as in the clear cell tumor. HNF-1beta expression was observed either in atypical endometriosis (four cases), or in endometriosis of a reactive nature (five cases). Furthermore, 16 of 40 (40%) endometriotic cysts without a neoplasm also expressed HNF-1beta, and the expression was almost exclusively observed in the epithelium showing inflammatory atypia. Our results indicate that HNF-1beta is an excellent molecular marker for ovarian clear cell tumors, including benign, borderline and malignant lesions. Early differentiation into the clear cell lineage takes place in ovarian endometriosis, not only in atypical endometriosis, but also in endometriosis with degenerative and regenerative changes, and this is probably responsible for the frequent occurrence of clear cell carcinoma in ovarian endometriosis.

Background: The aim of the present study was to evaluate microvessel density (MVD) in the cellular layers of ovarian endometriomata, with particular interest in the relationship with VEGF and survivin expressions by endothelial cells and with the diameter of the cysts., Materials and Methods: MVD and VEGF and survivin endothelial cell expressions were evaluated in 26 ovarian endometriotic cysts and correlated with the cyst diameter., Results: The mean MVD was higher in the inner specialized stroma of ectopic endometrium than in the outer fibrous capsule, but only in the fibrous capsule was MVD correlated with endothelial cell VEGF and survivin expressions as well as with the cyst diameter., Conclusion: The diameter of ovarian endometriotic cysts seems to be related to the angiogenic process involving the outer fibrous capsule, and not the inner specialized stroma of ectopic endometrium, since only in the capsule are vessels stimulated to proliferate by VEGF and protected from apoptosis by survivin, and their density is correlated to cyst diameter.

The aim of this study was to investigate whether a significant relation exists between the presence of a negative energy balance (NEB) in cows early in lactation and the reproductive performance after treatment with gonadotrophin-releasing hormone of cows with clinical cystic ovarian disease. Reproductive performance after treatment was assessed from the interval between treatment and first insemination (ITFI) and the interval between treatment and conception (ITC). Based on the outcome of the daily change of milk fat yield (Deltafatg) between the first and second milk recordings post-partum, cows were considered to have passed the NEB nadir (positive Deltafatg) or not (negative Deltafatg). Lactations (n = 430) were divided into four groups according to the interval between calving and first milk recording (ICMR): (i) 0-9 days; (ii) 10-19 days; (iii) 20-29 days; (iv) 30-49 days. The relation between a NEB-proxy parameter (Deltafatg) and reproductive performance was determined. A significant interaction existed for groups 1 and 3 (ICMR on 0-9 and 20-29 days post-partum respectively) with an increased and a decreased probability of being inseminated the first day after treatment (ITFI) respectively. However, no significant interaction was found between Deltafatg and the groups with regard to ITC.

Background: The objective of this study was to identify substantially increased proteins in bovine cystic follicular fluid (FF) in order to clarify the pathology and etiology of bovine ovarian follicular cysts (BOFC)., Methods: Proteins in normal and cystic FF samples were subjected to two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and were compared using silver stained gel images with PDQuest image analysis software. Peptides from these increased spots were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), and were identified based on the NCBI database by a peptide mass fingerprinting method., Results: Comparative proteomic analysis showed 8 increased protein spots present in cystic FF. MS analysis and database searching revealed that the increased proteins in cystic FF were bovine mitochondrial f1-atpase (BMFA), erythroid associated factor (EAF), methionine synthase (MeS), VEGF-receptor, glyceraldehydes 3-phosphate dehydrogenase (GAPDH), heat shock protein 70 (HSP70), beta-lactoglobulin (BLG) and succinate dehydrogenase Ip subunit (SD)., Conclusion: Our results suggest that these proteins are overexpressed in BOFC, and that they may play important roles in the pathogenesis of BOFC. Furthermore, these proteins in the FF could be useful biomarkers for BOFC.

Background: Glycodelin A levels in fluids of benign ovarian cysts, borderline tumours and ovarian cancer, as well as serum-levels of glycodelin A, were analysed in patients with benign and malignant ovarian tumours. The aim of the study was to investigate if the level of glycodelin A in body fluids of patients with malignant ovarian tumours could be a marker for the disease. Additionally, immunohistochemical investigations of glycodelin A expression in tissue of ovarian cancer were performed., Materials and Methods: A total of 158 samples of fluids from benign ovarian cysts, borderline tumours and ovarian cancer were collected during surgery in the Department of Obstetrics and Gynaecology at the University of Rostock, Germany. Additionally, 69 samples of serum from patients with benign ovarian cysts and ovarian cancer on the day before surgery were collected. An ELISA for determination of glycodelin A was used. Immunohistochemical detection of glycodelin A expression was performed on ovarian cancer tissue from 38 patients., Results: Malignant cystic fluids showed higher glycodelin A values (mean: 1814.4 ng/ml) compared to fluids of benign ovarian cysts (mean: 784.4 ng/ml). The results of glycodelin A determination were compared using the Mann- Whitney U-test for comparison of the means. There was a statistically significant difference between benign ovarian cysts and malignant ovarian cancer for the fluid (p <0.001). In addition, serum samples of malignant ovarian tumours also showed significantly higher glycodelin A values compared to serum levels of benign tumours (p<0.001). Immunohistochemical staining on ovarian cancer tissue showed a glycodelin A expression in 25-30% of the carcinoma cells., Conclusion: High levels of glycodelin A were found in cystic fluids of ovarian cancer. In addition, we also found higher levels of glycodelin A in the serum of patients with ovarian cancer compared to the serum of patients with benign ovarian cysts. Furthermore, ovarian cancer tissues showed intense staining with a glycodelin A antibody. Further investigations are necessary to show if glycodelin A quantification could help to diagnose ovarian cancer.

Objective: To investigate the expression of oxytocin receptor (OTR) in peritoneal and ovarian endometriotic lesions., Design: Retrospective nonrandomized study., Setting: University hospital endometriosis research center., Patient(s): Premenopausal women with histologically confirmed endometriosis were selected. Peritoneal endometriotic lesions (n = 120); ovarian endometriotic cysts (n = 40); peritoneal biopsies, distant from the endometriotic lesion (n = 55); and unaffected peritoneal biopsies from patients without endometriosis (n = 11) were obtained. Hysterectomy specimens from patients without endometriosis and/or adenomyosis were used for controls (n = 10)., Intervention(s): Histopathological examination of peritoneal and ovarian specimens for OTR expression and identification of smooth muscle cells by immunohistochemistry staining with antibodies against OTR and smooth muscle actin. In addition, Western blot analysis, double-immunofluorescence, and in vitro studies with primary cell cultures have been performed., Main Outcome Measure(s): Comparison of the immunoreactive score of the OTR and smooth muscle actin expression with the smooth muscle content in peritoneum with and without endometriosis., Result(s): In the epithelial cells of endometriotic lesions, we could demonstrate a high OTR expression. The stromal cells were OTR negative with the exception of some single cells. By using a monoclonal anti-smooth muscle actin antibody, these cells could be identified as intrastromal OTR-positive smooth muscle cells. The peritoneum of women with endometriosis shows a significantly higher smooth muscle content than the peritoneum of women without endometriosis. There were no significant differences between the smooth muscle content of active or inactive lesions and the stage of disease., Conclusion(s): Oxytocin receptor is expressed in smooth muscle cells and epithelial cells of peritoneal endometriotic lesions and ovarian endometriotic cysts. The inhibition of OTR by specific inhibitors might be a useful approach for the treatment of endometriosis-associated pain.

We have isolated an endo-beta-galactosidase designated E-ABase from Clostridium perfringens ATCC 10543 capable of liberating both the A trisaccharide (A-Tri; GalNAcalpha1-->3(Fucalpha1-->2)Gal) and B trisaccharide (B-Tri; Galalpha1-->3(Fucalpha1-->2)Gal) from glycoconjugates containing blood group A and B glycotopes, respectively. We have subsequently cloned the gene (eabC) that encodes E-ABase from this organism. This gene was found to be identical to the CPE0329 gene of C. perfringens strain 13, whose product was labeled as a hypothetical protein (Shimizu, T., Ohtani, K., Hirakawa, H., Ohshima, K., Yamashita, A., Shiba, T., Ogasawara, N., Hattori, M., Kuhara, S., and Hayashi, H. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 996-1001). Since the amino acid sequence of E-ABase does not bear detectable similarity to any of the 97 existing families of glycoside hydrolases, we have proposed to assign this unusual enzyme to a new family, GH98. We also expressed eabC in Escherichia coli BL21(DE3) and obtained 27 mg of fully active recombinant E-ABase from 1 liter of culture. Recombinant E-ABase not only destroyed the blood group A and B antigenicity of human type A and B erythrocytes, but also released A-Tri and B-Tri from blood group A(+)- and B(+)- containing glycoconjugates. The structures of A-Tri and B-Tri liberated from A(+) porcine gastric mucin and B(+) human ovarian cyst glycoprotein were established by NMR spectroscopy. The unique specificity of E-ABase should make it useful for studying the structure and function of blood group A- and B-containing glycoconju-gates as well as for identifying other glycosidases belonging to the new GH98 family.

Currently available serum biomarkers are insufficiently reliable to distinguish patients with epithelial ovarian cancer (EOC) from healthy individuals. Metabonomics, the study of metabolic processes in biologic systems, is based on the use of (1)H-NMR spectroscopy and multivariate statistics for biochemical data generation and interpretation and may provide a characteristic fingerprint in disease. In an effort to examine the utility of the metabonomic approach for discriminating sera from women with EOC from healthy controls, we performed (1)H-NMR spectroscopic analysis on preoperative serum specimens obtained from 38 patients with EOC, 12 patients with benign ovarian cysts and 53 healthy women. After data reduction, we applied both unsupervised Principal Component Analysis (PCA) and supervised Soft Independent Modeling of Class Analogy (SIMCA) for pattern recognition. The sensitivity and specificity tradeoffs were summarized for each variable using the area under the receiver-operating characteristic (ROC) curve. In addition, we analyzed the regions of NMR spectra that most strongly influence separation of sera of EOC patients from healthy controls. PCA analysis allowed correct separation of all serum specimens from 38 patients with EOC (100%) from all of the 21 premenopausal normal samples (100%) and from all the sera from patients with benign ovarian disease (100%). In addition, it was possible to correctly separate 37 of 38 (97.4%) cancer specimens from 31 of 32 (97%) postmenopausal control sera. SIMCA analysis using the Cooman's plot demonstrated that sera classes from patients with EOC, benign ovarian cysts and the postmenopausal healthy controls did not share multivariate space, providing validation for the class separation. ROC analysis indicated that the sera from patients with and without disease could be identified with 100% sensitivity and specificity at the (1)H-NMR regions 2.77 parts per million (ppm) and 2.04 ppm from the origin (AUC of ROC curve = 1.0). In addition, the regression coefficients most influential for the EOC samples compared to postmenopausal controls lie around delta3.7 ppm (due mainly to sugar hydrogens). Other loadings most influential for the EOC samples lie around delta2.25 ppm and delta1.18 ppm. These findings indicate that (1)H-NMR metabonomic analysis of serum achieves complete separation of EOC patients from healthy controls. The metabonomic approach deserves further evaluation as a potential novel strategy for the early detection of epithelial ovarian cancer., ((c) 2004 Wiley-Liss, Inc.)

Unlabelled: The biological behaviour (conduct) of benign ovarial cysts is interesting due to their comparatively high frequency of appearance and clinical manifestation. (During the last five years their frequency is between 62%-65% of the gynaecological surgical interventions). With this current survey the authors try to set up a target for research study of the wall receptive status of benign ovarial cysts. This study should be treated as a start up point rendering a prognosis for their eventual biological development., Materials and Methods: An immunohistochemical survey was carried on with 50 patients who have already reached the surgical treatment in retrospective order for the period 2001-2003 (three years' period). The survey search is in direction receptive analysis of Estrogene /E2/ and Progesterone /P/ wall receptor in the ovarial cyst. A perspective observation was also carried on (control group of 50 cases) of hormonal-gestagene treatment of the benign ovarial cystogenesis. It appears to be the necessary prove material for hormonal receptors' participation in the cyst wall as an active factor for its development and involution., Results: In the basic group /surgical treatment/ it was stated: In 39 patients--E2/-/, P/-/; in 4 patients--E2/+/, P/+/ In 5 patients--E2/+/, P/-/; in 2 patients--E2/-/, P/+/, Conclusion: The executed retrospective analysis demonstrates statistical importance in receptor competency lack in the wall of benign ovarial cysts as the probable grounds for them not being influenced during the conservative treatment method. This method led to the surgical elimination of the cyst. The statistical reliability, connected with the receptor E2 participation in the therapeutic behaviour of the benign ovarial cysts, is not stated. The final conclusions acquired from the executed research work would be a discussion issue after the correlative aspects in the prospective group are surveyed. The benign ovarial cysts' hormonal profile would be taken into consideration also.

Objectives: The aim of this work was to determine gonadotropin (LH, FSH) levels in serum and cyst fluid in various type of ovarian epithelial neoplasms (benign, borderline, malignant) and to compare them with levels in benign cysts. Additionally we decided to estimate if there were some significant correlations between serum and ovarian cyst fluid in gonadotropin levels in all investigated groups., Materials and Methods: The study group included 74 patients before (n=36) and after (n=38) menopause, divided into four groups depending on the histopathologic diagnosis. Serum and cyst fluid levels of LH and FSH were determined in all patients., Results: We found statistically significant differences concerning LH and FSH levels in serum and cyst fluid between malignant and borderline tumors, between malignant tumors and benign cystadenomas and between malignant tumors and non-neoplastic cysts. We also found statistically significant correlations between serum and cyst fluid as to FSH and LH levels in patients with borderline tumors. There were no statistically significant correlations between serum and cyst fluid as to FSH and LH levels in patients with malignant epithelial tumors (group 1) and in patients with benign cystadenomas (group 3)., Conclusions: High FSH and LH levels in cyst fluid of malignant epithelial tumors of the ovary are in line with the gonadotropin theory of tumor growth. Simultaneous determination of various types inhibin levels appears to be an interesting topic for our future research.

A 4-yr retrospective analysis of cases diagnosed as pelvic endometriotic cysts based on clinical, radiological, and cytomorphological features was collected and analysed. Out of 15 cases, only 3 cases showed the presence of endometrial glands/stroma in the aspiration smears. However, 12 of them showed the presence of oval/petal-shaped structures throughout the smear ranging in size from 40 to 70 micro in maximum dimension. These structures stained positive with periodic acid Schiff (PAS) stain. Immunostaining for type IV collagen showed strong positivity, indicating their origin from the basement membrane of the glands. The presence of these "collagenous bodies" in endometriotic cysts appears to be characteristic. However, this needs to be confirmed by prospective studies with a larger number of patients. The mechanism of formation of these collagenous bodies is uncertain.

Stringent regulation of LH secretion from the pituitary is vital to ovarian function in mammals. Two rodent models of LH hypersecretion are the transgenic LHbeta-C-terminal peptide (LHbetaCTP) and estrogen receptor-alpha (ERalpha)-null (alphaERKO) mice. Both exhibit ovarian phenotypes of chronic anovulation, cystic and hemorrhagic follicles, lack of corpora lutea, interstitial/stromal hyperplasia, and elevated plasma estradiol and testosterone. Because ERbeta is highly expressed in granulosa cells of the ovary, we hypothesized the intraovarian actions of ERbeta may be necessary for full manifestation of phenotypes associated with LH hyperstimulation. To address this question, we generated female mice that possess elevated LH, but lack ERbeta, by breeding the LHbetaCTP and ERbeta-null (betaERKO) mice. A comparison of LHbetaCTP, alphaERKO, and betaERKO(LHCTP) females has allowed us to elucidate the contribution of each ER form to the pathologies and endocrinopathies that occur during chronic LH stimulation of the ovary. alphaERKO ovaries respond to elevated LH by exhibiting an amplified steroidogenic pathway characteristic of the follicular stage of the ovarian cycle, whereas wild-type(LHCTP) and betaERKO(LHCTP) females exhibit a steroidogenic profile more characteristic of the luteal stage. In addition, the hemorrhagic and cystic follicles of the LHbetaCTP and alphaERKO ovaries require the intraovarian actions of ERbeta for manifestation, because they were lacking in the betaERKO(LHCTP) ovary. In turn, ectopic expression of the Leydig cell-specific enzyme, Hsd17b3, and male-like testosterone synthesis in the alphaERKO ovary are unique to this genotype and are therefore the culmination of elevated LH and the loss of functional ERalpha within the ovary.

The aim of this work was to compare mean concentrations of gonadotropins in serum and fluid from malignant and benign ovarian tumors. We enrolled 126 patients diagnosed with malignant epithelial tumors (n=40), borderline epithelial tumors (n=14), benign cystadenomas (n=28) and simple cysts (n=44) of the ovary. Premenopausal and postmenopausal subgroups were formed in each group. The concentration of FSH and LH was measured in serum and tumor fluid and the serum/tumor fluid ratio was calculated. The results in each group were compared and the sensitivity, specificity, positive and negative predictive values were determined. Mean concentrations of both gonadotropins in ovarian cancer fluid were significantly higher than in the remaining groups (P ranged from <0.005 to <0.0001). Mean serum/fluid ratios were lowest in ovarian cancer (FSH=2.91, LH=4.19). Our findings support the hypothesis that gonadotropins are involved in ovarian carcinogenesis and suggest that gonadotropin serum/tumor fluid ratios could be of value in the differential diagnosis of functional and organic cysts of the ovary., (Copyright 2004 Society for Endocrinology)

Objective: With neoplastic progression, the precursor of epithelial ovarian cancers, the ovarian surface epithelium (OSE), undergoes Mullerian differentiation, usually of the oviductal type. The aim of this study was to examine the expression of oviduct-specific glycoprotein (OGP), a marker of normal oviductal epithelium, for use as a diagnostic or prognostic marker for ovarian cancer., Methods and Materials: Immunohistochemical analysis for OGP was performed on 389 ovarian tumors and 19 normal ovaries, as well as 433 cases representing 45 normal tissues and 51 benign and malignant tumor types from 37 different tissues., Results: OGP was absent in OSE but present in 28 of 31 epithelial inclusion cysts, 13 of 14 (93%) serous cystadenomas, and 46 of 65 (71%) serous borderline tumors. Of 183 serous adenocarcinomas, 26 (14%) were positive for OGP, including 5 of 8 (63%) grade I, 7 of 41 (17%) grade II, and 14 of 134 (10%) grade III carcinomas. OGP was found in 7 of 14 (50%) borderline and 9 of 15 (60%) malignant mucinous ovarian tumors and in 10 of 39 (26%) endometrioid adenocarcinomas. The localization of OGP in the lumen of glandular structures suggested that it was secreted. OGP was absent in 41 of 45 normal tissues and positive in oviduct and, weakly, in salivary gland, duodenum, and ileum. Forty-six types of nongynecologic tumors were negative, as were gynecologic neoplasms except for 2 of 47 cervical and 3 of 56 endometrial carcinomas., Conclusion: OGP is a new tubal differentiation marker which characterizes benign and borderline serous neoplasms and may indicate early events in ovarian carcinogenesis.

Objectives: To investigate the expression of sulfatides in the tissue homogenates of malignant ovarian tumors, benign ovarian tumors, and control tissues and to study the relation between this marker and other clinico-pathological criteria such as the tumor type, grade of differentiation, surgical stage and ovulatory years., Design: Cross-sectional study., Setting: Department of Obstetrics and Gynecology and Department of Biochemistry, Assuit university hospital., Subjects: Forty-six patients had malignant ovarian tumors. Sixteen patients had benign ovarian neoplasm. Thirty patients, with normal ovaries, represented the control group., Methods: A sample of the tumor or from the normal ovary (the control group) was sent for histopathological and biochemical examination. Sulfatides were measured by a rapid and sensitive spectrophotometric method., Results: There was a significant rise in benign tumors [median and range 43 (38-53)], than in the control group, 21 (18-31), P-value = 0.000. In malignant tumors, the median value of sulfatides was significantly higher than in benign tumors [127 (71-193), P-value = 0.000]. Sulfatides were significantly higher in patients with more ovulatory years and tumors of advanced stages (stage III/IV) and poor differentiation., Conclusions: Sulfatides may play a role in the pathogenesis of benign and malignant ovarian tumors. It may also predict advanced stages in patients who are apparently early stage. It is also a candidate to study of their association with response to chemotherapy.

Several experimental models have been developed for the study of the polycystic ovarian syndrome in the rat. In the present study, the syndrome was induced by exposure to constant light, and the expression of cytoskeletal proteins in the follicular wall was evaluated by immunohistochemistry. We analyzed the immunohistochemically stained area (IHCSA) by image analysis to evaluate the expression of intermediate filaments (vimentin, desmin, cytokeratins, gliofibrillary acidic protein and neurofilaments) and alpha-smooth muscle actin (alpha-SMA) in cystic ovaries in relation to normal ovaries. The granulosa cell layer of cystic follicles had a significantly greater IHCSA for vimentin than the normal antral follicles. This difference was also significant between atretic and antral follicles. Cytokeratins showed a very low expression in the granulosa cells of antral follicles of control ovaries while in granulosa cells of atretic and cystic follicles they showed a significantly higher IHCSA. Immunohistochemical localization of desmin and alpha-SMA was restricted to the theca externa. Immunoreactivity for gliofibrillary acidic protein and neurofilament was negative. The highest intensity in the staining with vimentin and cytokeratins observed in the granulosa cells of the cystic follicles is probably due to structural and functional changes that occur during the process of cystogenesis and they could be associated with intense changes in the expression of cytoskeletal proteins that may be essential to the proper cellular functioning., (2004 S. Karger AG, Basel.)

Hepatocyte growth factor (HGF) is known to take role in oncogenesis and tumoral behavior of the tumors of the organs that contain mesenchymal and epithelial cells together. This study aims to compare HGF levels in cyst fluids of epithelial ovarian cancer and benign ovarian cysts and look for the role of HGF in ovarian carcinogenesis. Twenty-four consecutive patients with ovarian cancer and 34 with benign cysts of ovary were recruited prospectively at Gynecologic Oncology Departments of SSK Ankara Maternity Hospital and Hacettepe University School of Medicine between 2001 and 2002. Cyst fluids were collected during primary staging in cancer patients and during laparatomy for benign patients. HGF levels were measured by enzyme-linked immunosorbent assay method. Median HGF levels of the benign ovarian cysts and epithelial ovarian tumoral fluids were found as 3822 pg/ml (85-15,253 pgr/ml) and 12,962 pgr/ml (4136-16,025 pgr/ml), respectively. Malign cyst fluids have higher HGF levels when compared with benign ovarian cysts (P < 0.01). This finding suggests that HGF may take a paracrine role in oncogenic differentiation and tumoral development of epithelial ovarian cancers. Mechanisms that take role in HGF secretion and the answers of the neighboring epithelial cells to HGF during tumoral development need to be investigated.

Background: The authors suggested that the loss of collagen IV and laminin-containing basement membrane and the loss of Disabled-2 (Dab2) expression were two critical events associated with morphologic dysplastic changes of the ovarian surface epithelium as a step in tumorigenicity. Both the basement membrane and Dab2, a candidate tumor suppressor of ovarian carcinoma, were involved in epithelial cell surface positioning and organization. The authors speculated that the purging of the basement membrane may be similar to the proteolysis during gonadotropin-stimulated ovulation, a cyclooxygenase 2 (Cox-2)-mediated process., Methods: Prophylactic oophorectomy is used to prevent breast and ovarian carcinoma in high-risk populations. These ovarian tissue specimens often contain an increased presence of morphologically abnormal lesions that are believed to be preneoplastic. The authors evaluated archived prophylactic oophorectomy specimens to verify whether the loss of Dab2 expression and the removal of the basement membrane that occur at the ovarian surface and inclusion cyst epithelia are molecular markers of preneoplastic lesions. Of the 36 samples containing identifiable ovarian surface epithelial components on slides, immunostaining was employed to evaluate the intactness of the basement membrane (periodic acid-Schiff [PAS], collagen IV, and laminin) and the expression of Dab2 and Cox-2. Expression of Cox-1 and Cox-2 also were evaluated in cultured ovarian surface epithelial cells prepared from ovarian tissue specimens removed from patients who underwent prophylactic surgery., Results: The morphologically normal ovarian surface epithelium typically contained a collagen IV- and laminin-positive basement membrane, which also was detected by PAS staining. Many morphologically altered areas, such as papillomatosis, invaginations, inclusion cysts, stratification, adenomas, and microscopic adenocarcinomas, were found in these specimens. Both the morphologically altered and adjacent morphologically normal epithelia consistently exhibited loss of basement membrane and/or Dab2 expression and an increase in Cox-2 staining. Frequently, an increase in Cox-2 staining was correlated with the loss of epithelial basement membrane in morphologically normal areas., Conclusions: The loss of Dab2 and basement membrane and the overexpression of Cox-2 were observed in presumptive neoplastic precursor areas of oophorectomy specimens obtained from a population at high risk for ovarian carcinoma. Transient loss of collagen IV and laminin in the basement membrane of the preneoplastic epithelium and the loss of Dab2 expression are common early events associated with morphologic alteration and tumorigenicity of the ovarian surface epithelium. The authors concluded that Cox-2 overexpression may play a role in the purging of basement membrane of the ovarian surface epithelium, mimicking the process of ovulation. Further experiments may be able to test the hypothetical model derived from these histologic observations., (Copyright 2003 American Cancer Society.)

Ovarian cancer remains still associated with poor prognosis because it is diagnosed predominantly at advanced stages. Ovarian-specific tumor markers do not yet exist for early detection of the disease. At the search of diagnostic markers for ovarian cancer, proteomic-based approaches have focused on novel investigations of neoplastic processes in tumor patients. Cystic fluids of malignant and benign ovarian tumors and serum from the corresponding patients were collected and processed for two-dimensional gel electrophoresis. Proteins were visualized on the gels by silver staining. At the low molecular mass level between 10 and 20 kDa, selected protein spots were additionally processed for nanospray mass spectrometry and partial amino acid sequencing. For protein identification, the sequencing results were compared with computer information from a protein data bank. Protein patterns from cystic fluids of ovarian carcinomas differed significantly from those of benign cysts and revealed additional polypeptides at low molecular mass level between 10 and 20 kDa. Protein patterns from serum of patients with malignant ovarian tumors also contained additional polypeptides between 10 and 20 kDa that were not detected in serum from patients with benign cysts. The additional proteins in serum were present in similar electrophoretic positions compared with those found in the cystic fluid of the corresponding ovarian carcinomas. Protein spots in the range of 10-20 kDa were selected for partial amino acid sequencing. Two protein spots were identified as calgranulin A and three spots as calgranulin B. Either both proteins or only calgranulin A or B were present in cystic fluid from ovarian carcinomas and serum of the corresponding patients. These two proteins were absent or not detectable in fluid from benign ovarian cysts and in serum from those patients. Our investigations concerning protein patterns in cystic fluid of malignant and benign ovarian tumors provide new information about alterations in protein synthesis linked to neoplastic events of the ovary. With the proteomic strategy, new tumor markers are characterized and may serve for diagnostic purposes of patients with ovarian cancer.

Hepatocyte growth factor (HGF) is known to take a role in oncogenesis and tumoral behavior of the tumors of the organs that contain both mesenchymal and epithelial cells. This study compares HGF levels in cyst fluids of epithelial ovarian cancer and benign ovarian cysts to look for the role of HGF in ovarian carcinogenesis. Twenty-four consecutive patients with ovarian cancer and 34 with benign ovarian cysts were recruited prospectively at the Gynecologic Oncology Departments of SSK Ankara Maternity Hospital and Hacettepe University School of Medicine between 2001 and 2002. Cyst fluids were collected during primary staging in cancer patients and during laparatomy for benign patients. HGF levels were measured by ELISA method. Median HGF levels of the benign ovarian cysts and epithelial ovarian tumoral fluids were found to be 3822 pg/ml (85-15253 pg/ml) and 12962 pg/ml (4136-16025 pg/ml), respectively. Malignant cyst fluids have higher HGF levels when compared with benign ovarian cysts (P < 0.01). This finding suggests that HGF may take a paracrine role in oncogenic differentiation and tumoral development of epithelial ovarian cancers. Mechanisms that take a role in HGF secretion and the responses of neighboring epithelial cells to HGF during tumoral development need to be investigated.

Objectives: To determine cyst fluid and serum vascular endothelial growth factor (VEGF) concentrations in patients with ovarian masses and to investigate the efficiency of this modulator in the clinical management of cystic pelvic masses., Methods: Needle puncture for cyst fluid aspiration were performed on 88 cystic ovarian masses intraoperatively. Forty-five patients with benign and 43 patients with malignant ovarian pathology were analyzed for cyst fluid and serum VEGF concentrations. Both cystic fluid and serum VEGF concentration were determined by enzyme-linked immunosorbent assay (ELISA)., Results: Cyst fluid VEGF levels of malignant cysts (40.65+/-17.69 ng/ml) were significantly higher than those of benign cysts (12.53+/-6.13 ng/ml; P<0.001). Similarly, higher serum VEGF concentrations were found in patients with malignant disease (0.72+/-0.17 ng/ml) compared with benign cysts (0.33+/-0.11 ng/ml; P<0.001). A statistically significant correlation was observed between cyst fluid and serum VEGF levels in both malignant and benign cysts. For serum VEGF, at a cut-off value of 0.41 ng/ml; sensitivity, specificity, PPV, and NPV were 95%, 78%, 80% and 95%, respectively. No significant correlation between cyst fluid VEGF concentration and tumor stage or grade could be found., Conclusions: Significantly higher concentrations of VEGF are present in cyst fluid and serum of patients with malignant ovarian cysts compared with benign ovarian ones. There is no relation between VEGF and tumor stage or grade.

Unlabelled: Changes in the peritoneal fluid (PF) environment have been implicated in the pathogenesis of endometriosis as well as in the decrease of fertility., Objective: To evaluate the concentration of glutathione in PF of women with endometriosis., Patients: Twenty-one patients with endometriosis (I or II rAFS stage, n=11; III or IV rAFS stage, n=10), and 29 patients with follicular or dermoid ovarian cysts (n=17 and n=12, respectively)., Results: Mean (+/-S.D.) PF glutathione concentration was 0.22+/-0.01 micromol/ml in patients with minimal or mild endometriosis, 0.21+/-0.05 micromol/ml in women with III or IV stage of the disease, 0.24 +/- 0.03 micromol/ml in women with follicle ovarian cysts, and 0.23+/-0.05 micromol/ml in patients with dermoid tumors of ovaries. No significant difference in the peritoneal glutathione level was found between the groups., Conclusion: These results suggest that PF glutathione is not involved in the progression of endometriosis.

Modern gonadotrophin-releasing hormone (GnRH) antagonists such as cetrorelix and ganirelix reliably prevent premature LH surges in controlled ovarian hyperstimulation for assisted reproductive technologies. Cetrorelix and ganirelix are safe and effective compounds. Because of their distinct pharmacological mode of action, it has been possible to achieve a significant reduction of treatment time. Fertilization and pregnancy rates are comparable to those obtained in agonist protocols for ovarian stimulation. No allergic or hyperergic reactions have been reported, and patient compliance is excellent. The fact that GnRH antagonists allow an immediate suppression of gonadotrophin concentrations while preserving pituitary responsiveness to endogenous GnRH provides enormous flexibility in treatment. GnRH antagonists have helped to overcome some major disadvantages of GnRH agonists, especially of the long protocol, which is currently the standard protocol for ovarian stimulation. The mistaken administration of antagonists during early pregnancy is not possible, since they are only administered during ovarian stimulation when a premature LH surge may be imminent. They are used in the spontaneous cycle or after pretreatment with oral contraceptives. Pregnancy can easily be ruled out by testing for human gonadotrophic hormone before onset of gonadotrophin stimulation on the second or third day of the cycle. Since flare-up effects are absent, there is no risk of cyst formation. Hormonal withdrawal symptoms are eliminated, since no period of pituitary suppression occurs, and therefore exogenous gonadotrophins are not required. Overall, the duration of the stimulation cycle is as short as a normal menstrual cycle. The procedure seems to be safer than the long protocol, since the most serious complication, the occurrence of severe cases of OHSS, is reduced. It is also safe with respect to the course of pregnancies and the health of offspring. Both protocols developed so far, the single dose and multiple dose antagonist protocol, are comparable, utilizing data from the large prospective phase IIIb studies. Although several studies have indicated a slight reduction in pregnancy rate with GnRH antagonists as compared with agonists, this problem may be rectified by developing flexible antagonist regimens designed for individual patients. Introducing flexible GnRH antagonist regimens should be the area of research in the near future.

Objective: To compare immunohistochemical staining patterns in noncystic and cystic endometriosis lesions., Design: Experimental., Setting: Archived pathology material in an academic research environment., Patient(s): Endometriosis tissues from the pathology archives including slide tissue sections and blocks., Intervention(s): None; this was a retrospective study., Main Outcome Measure(s): Immunohistochemical staining of the tissues was performed using anti-bcl-2, anti-p53, anti-matrix metalloproteinase IX, and anti-collagen VI antibodies. Staining was qualitatively assessed in terms of extent and intensity., Result(s): p53 showed no staining in both groups. Anti-bcl-2 stained 100% (30/30) of endometriosis lesions compared with only 23% (7/30) of endometriotic cysts (P<.0001), and anti-matrix metalloproteinase IX stained 85% (23/27) of endometriosis lesions and only 39% (14/36) of endometriotic cysts (P=.0003). Anti-collagen VI, however, stained only 6% (2/35) of endometriosis lesions and 75% (21/28) of endometriotic cysts (P<.0001)., Conclusion(s): Compared with endometriosis lesions, endometriotic cysts display different expression of proteins with relative overexpression of collagen VI and underexpression of bcl-2 and metalloproteinase IX. This report is the first comparative immunohistochemical study showing these differences.

The aim of this study was to determine possible links between steroidogenic activity of single ovarian cysts and response to intramuscular treatment with 20 microg of buserelin (GnRH-analogue) after cyst emptying, in pluriparous Friesian cows bearing a singleton cyst treated not earlier than 55 days post-partum. Progesterone, 17beta-estradiol and testosterone were determined in cystic fluids collected by needle aspiration of the cyst. Of the cows, 75.6% began ovarian cyclicity within 30 days after treatment with a conception rate of 64.7%. In this study it was found that as progesterone concentration in cystic fluids rose, the number of positive responses to the treatment fell.

Background: A number of histologic and epidemiologic studies have suggested an association between endometriosis and ovarian carcinoma. Some reports have described a transition from endometriosis to atypical endometriosis to carcinoma. Using immunohistochemistry, the authors compared staining patterns in benign endometriotic cysts with ovarian tumors and the endometriotic cyst lining from which they arose, in an attempt to identify sequential or etiologic correlations., Methods: One hundred thirteen formalin-fixed, paraffin-embedded sections were studied (30 benign ovarian endometriotic cysts, 24 endometriotic cysts containing endometrioid carcinomas, 19 endometriotic cysts harboring clear cell carcinomas, and 40 ovarian papillary serous cystadenocarcinomas). All sections were immunostained with anti-bcl-2 and anti-p53 antibodies using the streptavidin-biotin method., Results: bcl-2 was reported to stain 23% of benign endometriotic cysts, 67% of endometrioid carcinomas, 73% of clear cell carcinomas, and 50% of papillary serous carcinomas. Approximately 42% of benign endometriotic lesions adjacent to the endometrioid carcinoma and 73% adjacent to clear cell carcinomas were found to stain for bcl-2 (p = 0.274 [not significant (NS)] and P = 0.008, respectively). p53 staining was negative in the benign endometriotic cyst group and was positive in 37-55% of the group with tumors. p53 staining was positive in 25% of the benign endometriotic lesions next to the endometrioid carcinoma and in 9% of the benign endometriotic lesions next to clear cell carcinoma (P = 0.014 and P = 0.239 [NS], respectively)., Conclusions: The results of the current study suggest that alterations in bcl-2 and p53 may be associated with the malignant transformation of endometriotic cysts., (Copyright 2002 American Cancer Society.)

Background: Changes in angiogenesis and expression of extracellular matrix-degrading enzymes have been substantiated in tumour changeover and progression., Methods: Tissues from 44 biopsies of stage III and IV ovarian endometriomas, and 10 biopsies of normal (control) endometrium were investigated immunohistochemically to count microvessels, and by in situ hybridization to assess the expression of mRNA of matrix metalloproteinase-2 (MMP-2) and MMP-9. Implants of the tissues were investigated in the chick embryo chorioallantoic membrane (CAM) assay to determine their angiogenic capacity., Results: The endometriomas displayed significantly higher counts than normal endometria and the highest values were associated with the deepest invasion level (stage IV). Microvessels localized in close association with ectopic endometrial cells in the form of winding and arborized tubes, often dilated in microaneurysmatic segments. These were absent in normal endometrium. Expression of MMP-2 and MMP-9 mRNA, evaluated as percentages of positive biopsies and intensity of expression, was up-regulated in endometriomas and more pronounced in stage IV. MMP-2 and MMP-9 mRNA were also expressed by host stromal cells, including microvascular endothelial cells, fibroblasts and macrophages, whereas the control endometrium showed very little expression of MMP-2 mRNA in a few endothelial cells and no expression of MMP-9 mRNA. Implants from stage IV endometrioma induced a more intense vasoproliferative response than those from stage III, while no vasoproliferative response was induced by the normal endometrium., Conclusion: These data suggest that angiogenesis and degradation of extracellular matrix occur together in endometriosis and are more pronounced in stage IV, and that endometriosis cells and some host stromal cell populations co-operate in disease progression.

Tumor markers have been investigated in differentiation of benign and malignant tumors. We analyzed CA 125 and vascular endothelial growth factor (VEGF) levels in serum and cyst fluid in patients with epithelial ovarian tumors. Serum and tumor cyst fluid of 50 patients with ovarian epithelial tumors (7 malignant, 3 bordeline and 40 benign) were assayed for VEGF by ELISA and CA 125 levels by chemoluminescence. CA 125 serum levels were significantly higher in patients with malignant and borderline tumors than in patients with benign cysts (p = 0.0005). CA 125 cyst fluid contents were comparable for malignant, borderline and benign ovarian tumors (p = 0.39). Significantly higher levels of VEGF were present in cyst fluid for malignant and borderline tumors compared with benign cysts (p < 0.0001); however, serum levels of VEGF were similar among all patients (p = 0.25). The CA 125 serum levels correlated with matched VEGF cyst fluid levels (r = 0.44, p = 0.0015). Serum CA 125 and cystic VEGF were good methods to differentiate benign and malignant epithelial ovarian tumors. Patients with elevated intracystic VEGF levels presented significantly higher CA 125 serum levels, although the CA 125 intracystic content overlapped. The angiogenesis and enhancement of vascular permeability induced by VEGF represents a new hypothesis for the release of the CA 125 antigen into the circulation in patients with ovarian epithelial neoplasm., (Copyright 2002 S. Karger AG, Basel)

Owing to the expression of more than one type of galectin in animal tissues, the delineation of the functions of individual members of this lectin family requires the precise definition of their carbohydrate specificities. Thus, the binding properties of chicken liver galectin (CG-16) to glycoproteins (gps) and Streptococcus pneumoniae type 14 polysaccharide were studied by the biotin/avidin-mediated microtitre-plate lectin-binding assay and by the inhibition of lectin-glycan interactions with sugar ligands. Among 33 glycans tested for lectin binding, CG-16 reacted best with human blood group ABO (H) precursor gps and their equivalent gps, which contain a high density of D-galactopyranose(beta1-4)2-acetamido-2-deoxy-D-glucopyranose [Gal(beta1-4)GlcNAc] and Gal(beta1-3)GlcNAc residues at the non-reducing end, but this lectin reacted weakly or not at all with A-,H-type and sialylated gps. Among the oligosaccharides tested by the inhibition assay, the tri-antennary Gal(beta1-4)GlcNAc (Tri-II) was the best. It was 2.1x10(3) nM and 3.0 times more potent than Gal and Gal(beta1-4)GlcNAc (II)/Gal(beta1-3)GlcNAc(beta1-3)Gal(beta1-4)Glc (lacto-N-tetraose) respectively. CG-16 has a preference for the beta-anomer of Gal at the non-reducing end of oligosaccharides with a Gal(beta1-4) linkage >Gal(beta1-3)> or =Gal(beta1-6). From the results, it can be concluded that the combining site of this agglutinin should be a cavity type, and that a hydrophobic interaction in the vicinity of the binding site for sugar accommodation increases the affinity. The binding site of CG-16 is as large as a tetrasaccharide of the beta-anomer of Gal, and is most complementary to lacto-N-tetraose and Gal(beta1-4)GlcNAc related sequences.

Objective: To determine the levels of the angiogenic factors vascular endothelial growth factor (VEGF) and interleukin (IL-8) in ovarian cysts., Design: Prospective descriptive study., Setting: University hospital., Patient(s): One hundred women, of whom 9 had ovarian carcinomas, 38 had ovarian endometriomata, 43 had serous ovarian cysts, and 10 had follicular ovarian cysts., Intervention(s): Sampling of serum and ovarian cystic fluid before and during surgery., Main Outcome Measure: Levels of VEGF and IL-8 in cystic fluid and serum., Result(s): Levels of both VEGF and IL-8 were found to be significantly higher in the cystic fluid of ovarian carcinomas and endometriomata than in serous and follicular cysts. In endometriomata fluid, levels of VEGF and IL-8 were found to be directly correlated (r = 0.68; P=.0074). Serum levels of VEGF were significantly higher in women with ovarian carcinomas and endometriomata than in those with serous and follicular cysts. Ovarian cancers and endometriomata were similar in terms of cystic concentrations of VEGF and IL-8 and in serum levels of VEGF., Conclusion(s): An increase in angiogenic factors that differentiate ovarian carcinomas and endometriomata from other kinds of ovarian pathology is demonstrated.

53 women with an ovarian chocolate cyst were operated in a laparoscopy. A histopathological examination showed 60.4% of endometriomas and 39.6% of hemorrhagic lutein cysts. The average age of patients with endometriomas was 33.9 while of the ones with hemorrhagic lutein cysts was 28.6. The operated patients, who were under 35 years old, could be divided into halves, one half of them had endometriomas while the other half had hemorrhagic lutein cysts. However, endometriomas appeared significantly more often in the group of operated patients who were older than 35 years old.

Increased concentrations of TGF-beta 1 in endometriotic tissue are considered important in the pathophysiology of endometrial diseases since TGF-beta 1 may inhibit natural killer activity and induce angiogenesis and proliferation of endometrial stromal cells. In the present study we report on TGF-beta 1, IGF-1 and their receptor localization, as detected by Northern hybridization and immunohistochemistry, in ovarian endometriotic tissues removed during surgical procedures. We detected comparable expression of IGF-1 and IGF-1 receptor in the stromal and epithelial compartments, thus confirming disregulated expression of the IGF system in ovarian endometriosis. On the contrary, strongly increased TGF-beta 1 steady state level mRNA expression was detected in all endometriotic samples. In addition, we demonstrated weak TGF-beta 1 immunohistochemical expression in the epithelial lining and intense expression in the cellular stroma of ovarian endometriomas, thus suggesting that TGF-beta 1 could have an important role in the maintenance and propagation of the disease. On the basis of these preliminary results we can assume that TGF-beta 1, IGF-1 and their receptors may play an important role in the pathogenesis of endometriosis.

Objective: To evaluate the expression of inhibin A and activin A in ovarian endometriosis., Design: Uncontrolled cross-sectional study and controlled prospective in vitro study., Setting: Academic health centers in Siena, Udine, Sassari, and Milan, Italy., Patient(s): A group of women (n = 19) who underwent laparoscopic excision of ovarian endometriotic cysts., Intervention(s): Specimens of serum, peritoneal fluid, and cystic fluid, ovarian tissue for immunohistochemistry, and endometriotic cells for primary culture were collected. Cell cultures were also prepared from proliferative endometrium of women without endometriosis., Main Outcome Measures: Dimeric inhibin A and activin A concentrations in biological fluids; immunostaining of alpha and betaA subunits in ovarian endometrioma; alpha and betaA gene expression in cultured endometriotic cells compared with normal endometrium., Result(s): Inhibin A and activin A concentrations in the cystic fluid were slightly higher than in peritoneal fluid and significantly higher than in serum (P<.05). Immunoreactive alpha and betaA subunits were strongly expressed both in the epithelial and stromal components of ovarian endometrioma. The relative abundance of betaA mRNA was significantly decreased in endometriotic cells compared with eutopic stromal cells., Conclusion(s): The results of the present study provide evidence for a local production and secretion of inhibin A and activin A in ovarian endometriotic cysts.

Reports of supernumerary ovaries are rare. We describe two such cases, one with fibroma and the other with endometriosis and cystic change. A large fibroma measuring 17.4 x 12.0 x 7.5 cm in size was found in the supernumerary ovary of the omentum in the first case of a 47-year-old married woman with Meig's syndrome. The second case was associated with endometriosis and cystic change, measuring 11 x 5 x 3 cm in size and located in the upper abdominal cavity. It was attached to the uterus of a 28-year-old pregnant woman who had neither fibroma nor Meig's syndrome. Histologically, corpus albicans and a few primordial germ cells were demonstrated, respectively. A fibroma showing a storiform pattern was found in the first case. The second case had endometriosis and a thin-walled cyst with bleeding and necrosis caused by torsion. Immunohistochemically, desmin, alpha-smooth muscle actin, c-kit, CA125, Na+/K+ATPase, overexpression of p53, myc and ras were all negative in the fibroma cells of the first case, and in the endometriosis and cyst wall of the second case. The fibroma cells were positive for vimentin and estrogen receptor, and the proliferating cell nuclear antigen was sporadically demonstrated in their nuclei. The mutation of the p53 gene at exons 5-8 was not detected by sequence analysis. Using RT-PCR, bax, bcl-2 and p16 were not detected either. Clinically, the two cases presented here did not show abnormal hormonal symptoms. They were diagnosed as abdominal tumors or masses. Based on these considerations, one might assume that supernumerary ovaries are probably more frequent than reported at present.