Ovarian Ascites Research Articles

The influence of aquaporin-1 and microvessel density on ovarian carcinogenesis and ascites formation

This study aimed at investigating aquaporin-1 (AQP1) distribution and expression in primary ovarian epithelial tumors, correlating with clinicopathologic variables and intratumoral microvessel density (IMD). The AQP1 expression and IMD in 105 cases with primary epithelial ovarian tumors were measured by semiquantitative immunohistochemical technique. AQP1 was located mainly in microvessels and small vessels but seldom in tumor cells. Expression of AQP1 and IMD in ovarian malignant tumors was significantly higher than that in borderline tumors (P= 0.000, P= 0.001, respectively), and that in borderline tumors was higher than in benign tumors (P= 0.008, P= 0.028, respectively). The expression of AQP1 in FIGO stage III-IV was more than that in stage I-II (P= 0.001), and cases with ascites volume greater than 1000 mL were more than cases with ascites volume less than 1000 mL (P= 0.000). There is a positively correlated relationship between expression of AQP1 and IMD (correlation coefficient 0.60, P= 0.000) and between expression of AQP1 and ascites volume (correlation coefficient 0.57, P= 0.000). These data implicate that high AQP1 expression may play an important role in the ovarian carcinogenesis, progression, and ascites formation. Further studies into the mechanism of AQP1 regulation and the relationship between AQP1 expression and tumor angiogenesis may lead to novel therapies for ovarian carcinoma.

Elevated CA125 level associated with Meigs' syndrome: case report and review of the literature

Meigs' syndrome is the association of ovarian fibroma, pleural effusion, and ascites. Meigs' syndrome with marked elevation of CA125 is an unusual clinical condition reported in 27 cases in the literature. The patient was a 46-year-old woman with right pleural effusion, ascites, ovarian tumor, and CA125 level of 1808 U/mL. Tomography revealed ascites and bilobate pelvic tumor of approximately 25 cm. The diagnosis of advanced epithelial ovarian cancer was considered, and the patient was treated with chemotherapy. Three chemotherapy schemes were applied due to the total lack of response in tumor volume; however, CA125 decreased to 90 U/mL. Thus, surgery was performed with resection of 25 cm of the left ovarian tumor, with intact capsule and without implants; the result of histopathologic analysis was fibroma. Postoperative CA125 was 11 U/mL. Patients with elevated CA125 and ascites cytology positive for malignancy must be cautiously treated due to the possibility of false positives, even if the probability is low. Therefore, minimally invasive surgery for biopsy collection must be considered. Although the association between ovarian tumor, pleural effusion, ascites, and marked elevation of CA125 is highly indicative of epithelial ovarian cancer, Meigs' syndrome must be considered in the differential diagnosis.

Elevated CA125 level associated with Meigs’ syndrome: case report and review of the literature

Meigs' syndrome is the association of ovarian fibroma, pleural effusion, and ascites. Meigs' syndrome with marked elevation of CA125 is an unusual clinical condition reported in 27 cases in the literature. The patient was a 46-year-old woman with right pleural effusion, ascites, ovarian tumor, and CA125 level of 1808 U/mL. Tomography revealed ascites and bilobate pelvic tumor of approximately 25 cm. The diagnosis of advanced epithelial ovarian cancer was considered, and the patient was treated with chemotherapy. Three chemotherapy schemes were applied due to the total lack of response in tumor volume; however, CA125 decreased to 90 U/mL. Thus, surgery was performed with resection of 25 cm of the left ovarian tumor, with intact capsule and without implants; the result of histopathologic analysis was fibroma. Postoperative CA125 was 11 U/mL. Patients with elevated CA125 and ascites cytology positive for malignancy must be cautiously treated due to the possibility of false positives, even if the probability is low. Therefore, minimally invasive surgery for biopsy collection must be considered. Although the association between ovarian tumor, pleural effusion, ascites, and marked elevation of CA125 is highly indicative of epithelial ovarian cancer, Meigs' syndrome must be considered in the differential diagnosis.

Functional duality of merlin: A conundrum of proteome complexity

Merlin and ezrin proteins belong to the same gene family, i.e., MERM containing merlin, ezrin radixin and moesin. Members of this family possess an extensive homology in their amino acid sequences. It has been shown that merlin is a tumor suppressor, while ezrin is a promoter in tumor progression. The expression of these two proteins is commonly found inversely proportional to each other in cancer cells, i.e. down-regulation of merlin concomitant with over-expression of ezrin and vice versa. However, when determining merlin and ezrin in ovarian carcinoma cells, we have observed that both merlin and ezrin could be over-expressed simultaneously in some ovarian cancer (OVCA) cell lines and OVCA ascites cells, suggesting that merlin could be an oncoprotein rather than a tumor suppressor protein in certain OVCA cells. The functional duality of merlin might represent a paradigm in proteome complexity and is especially important in investigating multifactorial diseases such as cancer.

Plasma Levels of Soluble Vascular Endothelial Growth Factor Receptor-1 (sVEGFR-1) Play a Key Role in the Onset of Early (E-) and Late (L-) Ovarian Hyperstimulation Syndrome (OHSS)

OHSS is associated with ovarian stimulation in infertile women. E-OHSS is the consequence of hCG administration; L-OHSS is induced by trophoblastic hCG. Both develop in women carrying characteristics still not defined, and have an auto-regulation never explored before. VEGF is a system of ligands and receptors associated with increased vascular permeability (VP) in OHSS. Alternative mRNA splicing of its receptor 1, produces sVEGFR1 a natural inhibitor which reduces the availability of free (f-)-VEGF. We aimed to analyze the dynamics of the VEGF system in E- and L-OHSS in order to understand who develops OHSS, and the mechanism(s) determining its life span. Moreover, the only proposed VEGF sequester, (α2Macroglobulin (α2M), was also measured. A prospective study of women at risk (multicystic antral follicles in basal conditions and >20 oocytes retrieved) who did and did not develop OHSS, and patients without risk. Additionally, oocyte donation (OD) was compared to IVF-ICSI pregnancies, to determine the origin of the molecular players of L-OHSS E-OHSS: Non-pregnant patients were included. Group 1 (n=11) had no risk of OHSS; Group 2 (n=18) were women at risk who did not develop OHSS; Group 3 (n=8) did develop severe OHSS with culdocentesis due to ascites. Evaluation including blood parameters and pelvic ultrasound to evaluate ovarian size and ascites, was performed the day of ovum pick-up (day 0), 3, 6 10 and 14 days later. Serum and/or plasma was obtained. L-OHSS: Single pregnancies were studied weekly (4th to 12th) classified in: Women who did not develop (group 4, n=8); and did develop (group 5, n=4) severe OHSS requiring culdocentesis. An additional control group (n=4) was formed of single clinical pregnancies after OD and followed similar to groups 4 and 5, grouped in a single group named IVF-ICSI for comparison.Total VEGF, f-VEGF and sVEGFR1 were measured in plasma by ELISA . Serum E2 and P4 were measured by enzyme immunoassay, (α2M was measured employing nephelometry. Statistical methods were applied with the SPSS program; a p<0.05 was statistically significant. The figure shows the dynamics of sVEGFR1 secretion in E- and L-OHSS as well as comparison between IVF-ICSI and OD. The difference in the availability of sVEGFR1 determined the appearance of severe OHSS. Women in group 3 secreted very low levels as compared to groups 1 and 2. The placenta is an important source of sVEGFR1, but OD pregnancies are deficient in this regard. The differences in sVEGFR-1 determined the levels of f-VEGF in the luteal phase (E-OHSS) and first trimester of pregnancy (L-OHSS), whereas (α2M had no effect. Despite significantly (p<0.05) higher f-VEGF levels in OD, these patients did not develop OHSS.Analysis of serum E2 and P4 in the luteal phase showed levels compatible with a functional corpus luteum in group 3. Rather, an increase in sVEGFR1 may be the initiator of OHSS demise. In E- and L-OHSS, the ability to secrete sVEGFR1 reducing the availability of f-VEGF is the determinant factor which induces the severity and limits in time OHSS. f-VEGF increases VP in the ovaries, since OD patients do not develop OHSS.

PDGFR is activated through an autocrine mechanism in ovarian cancer

5073 Background: We demonstrated PDGFRα expression in ovarian cancer cells and in ovarian tumors using microarray hybridization and immunohistochemistry (Matei, Oncogene, 2002 and Matei, Clin Cance...

Cleavage of L1 in Exosomes and Apoptotic Membrane Vesicles Released from Ovarian Carcinoma Cells

The L1 adhesion molecule (CD171) is overexpressed in human ovarian and endometrial carcinomas and is associated with bad prognosis. Although expressed as a transmembrane molecule, L1 is released from carcinoma cells in a soluble form. Soluble L1 is present in serum and ascites of ovarian carcinoma patients. We investigated the mode of L1 cleavage and the function of soluble L1. We used ovarian carcinoma cell lines and ascites from ovarian carcinoma patients to analyze soluble L1 and L1 cleavage by Western blot analysis and ELISA. We find that in ovarian carcinoma cells the constitutive cleavage of L1 proceeds in secretory vesicles. We show that apoptotic stimuli like C2-ceramide, staurosporine, UV irradiation, and hypoxic conditions enhance L1-vesicle release resulting in elevated levels of soluble L1. Constitutive cleavage of L1 is mediated by a disintegrin and metalloproteinase 10, but under apoptotic conditions multiple metalloproteinases are involved. L1 cleavage occurs in two types of vesicles with distinct density features: constitutively released vesicles with similarity to exosomes and apoptotic vesicles. Both types of L1-containing vesicles are present in the ascites fluids of ovarian carcinoma patients. Soluble L1 from ascites is a potent inducer of cell migration and can trigger extracellular signal-regulated kinase phosphorylation. We suggest that tumor-derived vesicles may be an important source for soluble L1 that could regulate tumor cell function in an autocrine/paracrine fashion.

A Long Surviving Case of Pseudomeigs' Syndrome Caused by Krukenberg Tumor of the Stomach

Meigs syndrome is defined as the triad of benign ovarian tumor with ascites and pleural effusion that resolve after resection of the tumor. Pseudomeigs' syndrome is a serious disease characterized by malignant ovarian tumor, but ascites and hydrothorax usually reveal no malignant cells. Here, we report a 47-year-old pre-menapausal female patient with cardia cancer. Nearly 14 months after D3 dissection, she developed Krukenberg tumors on both ovaries causing a Pseudomeigs' syndrome with benign ascites and right hydrothorax, which resolved dramatically after resection of the tumors and rectouterine pouch peritonectomy. She survived nearly 3 years after metastasectomy with a total survival of 46 months. The patient died because of massive liver metastases. The present case suggests that Pseudomeigs' syndrome should be considered in patients with Krukenberg tumors, ascites and hydrothorax and that resection of the tumors may bring long-term palliation.

Assessment of prognostic factors in common ovarian tumors of varying malignancy

The outcome and prognosis of apparently similar cases of epithelial ovarian cancers with the same histology and stage is highly variable. The objective was to compare survival and prognostic factors of patients treated at the 1st Department of Obstetrics and Gynecology of Semmelweis University between 1993-2003 with the similar data of the 25th Annual Report on the Results of Treatment in Gynecological Cancer of FIGO. In addition, the aim was to assess the prognostic value of MMP activities and fibronectin concentration in ovarian tumour patients. The 25th Annual Report of FIGO included 5694 patients with ovarian tumours from 32 countries diagnosed and treated between 1995 and 1998. Hungary did not participate in this report. Between 1993 and 2003, 180 patients with common epithelial ovarian tumours had been treated at the 1st Department of Obstetrics and Gynecology of Semmelweis University. Treatment and survival data derived from medical record review and from the Population Register Office. In order to compare different prognostic factors, a multivariate Cox proportional regression analysis was performed. The authors measured MMP activities in 33 surgically removed ovarian tumours, serum and ascites by applying zymographic technique. Fibronectin content was determined by Western blot analysis and quantitated by densitometry. The 5-year survival was 90.0% and 30.9% in the case of ovarian tumours with low malignant potential and of epithelial ovarian cancers, respectively. Multivariate analysis identified adverse prognostic factors including advanced age (>60 years) and stage, high grade and suboptimal operation with residual macroscopic disease and the presence of ascites. However, the histological type was not identified to be an adverse prognostic factor in this study. No correlation could be seen between the histology of the ovarian tumours and the elevation of MMP-2/9 activity. More interestingly, however, MMP-9 expression and fibronectin concentration were significantly elevated and the activated forms of both MMP-9 and MMP-2 were more frequent in ovarian cancer patients who developed recurrent disease. A great deal of effort should be devoted to identification of further prognostic factors to improve treatment of ovarian cancer. These prognostic factors might help to identify those ovarian cancer patients at the time of diagnosis whose disease will have unfavourable outcome. Our data support the notion that high expression of MMP-9 and fibronectin indicates poor prognosis of ovarian cancer patients.

Proinvasive properties of ovarian cancer ascites-derived membrane vesicles.

Malignant ovarian ascites are rich in cellular components, membrane-bound vesicles, and soluble proteins. This study focused on the structure of membrane-bound vesicles and their ability to promote invasion in cultured malignant ovarian epithelium. Membrane vesicles were derived from women with stage I-IV malignant ovarian ascites and from nonmalignant gynecologic ascites. Isolated vesicles were characterized by immunofluorescence and Western blot analysis. Using gel zymography for matrix metalloproteinase (MMP) detection and a colorimetric assay for urokinase-type plasminogen activator (uPA) analysis, we analyzed the proteinase activities of MMP-2, MMP-9, and uPA from the prepared vesicles, whole cells isolated from ascites, and the cell-free ultracentrifuged supernatant. The invasiveness of established cultured malignant ovarian epithelium on addition of ascites-derived vesicles was tested using a Matrigel-based invasion assay. Fractionation of malignant ascites revealed that extracellular matrix-degrading proteinases including MMPs and uPA are localized preferentially in membrane vesicles. All malignant vesicles tested, regardless of cancer stage, stimulated invasion. Furthermore, the combination of ovarian cancer cells and membrane vesicles resulted in greater uPA activation than that of cells or vesicles alone. Membrane vesicles from malignant ascites were also found to contain activated MMP-2, MMP-9, and uPA. Our data suggest that vesicle-stimulated proteinase activation leads to increased extracellular matrix degradation, which may facilitate tumor cell invasion and metastasis.

Chemosensitivity and radiosensitivity profiles of four new human epithelial ovarian cancer cell lines exhibiting genetic alterations in BRCA2, TGFbeta-RII, KRAS2, TP53 and/or CDNK2A.

To address the cellular basis for the response to ovarian cancer treatment, we characterized the chemosensitivity and radiosensitivity of four human epithelial ovarian cancer cell lines that harbor different genetic alterations. The TOV-21G, TOV-81D, OV-90, and TOV-112D cell lines were derived from ovarian tumors (TOV) or ascites (OV) from chemotherapy- and radiotherapy-naive patients and were characterized by their mutation spectrum of BRCA2, TGFbeta-RII, KRAS2, TP53, and CDKN2A. Cells were monitored for survival following exposure at various concentrations to different cytotoxic agents including cisplatin, camptothecin or paclitaxel or to different doses of gamma-irradiation. At the lowest doses, the TGFbeta-RII-mutated and KRAS2-mutated cell line, TOV-21G, and the BRCA2-mutated cell line, TOV-81D, demonstrated a significantly higher sensitivity to cisplatin and gamma-irradiation than the TP53-mutated cell lines, TOV-112D and OV-90. At higher doses, differences between the TP53-mutated lines were observed with TOV-112D being less sensitive to cisplatin than OV-90 that also harbors a CDNK2A mutation. All cell lines were similarly sensitive to high doses of gamma-irradiation. In contrast, sensitivity to camptothecin or paclitaxel was not significantly different between all cell lines, irrespective of the mutation status of BRCA1, BRCA2, TGFbeta-RII, KRAS2, TP53, and CDKN2A. The observed responses to treatment are consistent with the current knowledge concerning BRCA2, TGFbeta-RII, KRAS2, TP53, and/or CDKN2A aberrant function.

Pseudo-Meigs' syndrome associated with malignant struma ovarii: a case report

Background. Malignant struma ovarii is a rare tumor, consisting of a struma ovarii with malignant transformation. The association of a malignant struma ovarii with pseudo-Meigs' syndrome and elevated Ca-125 levels has been never reported. Case. A 66-year-old woman presented with monolateral ovarian mass, ascites, hydrothorax, and elevated Ca-125 levels. Optimal medical staging was performed. Definitive histological examination revealed a malignant struma ovarii. The immediate and complete resolution of symptoms and rapid decline of both Ca-125 and thyroglobulin levels to normal value were achieved post-operatively. After counseling, strict follow-up was planned, and no adjuvant therapy was administered. Conclusion. We report the first case of pseudo-Meigs' syndrome associated with malignant struma ovarii and elevated Ca-125 levels. The choice of not performing adjuvant therapy is feasible after optimal surgery and adequate staging procedure given to the usually clinical benign course and the low incidence of metastases in malignant struma ovarii. Careful patient counseling is required.

Bilateral ovarian fibromatosis presenting with ascites and hirsutism

Background. Ovarian fibromatosis is a very rare nonneoplastic disease. Due to the rarity and atypical clinical presentations, they may give rise to a misdiagnosis of malignancy and unnecessary extensive surgical interventions. Literature lacks definitive data about this rare disease and its preoperative evaluations. MRI together with the intraoperative frozen section may help us to define the benign nature of the disease. In this report, we aimed to review the literature and give a highlight to the gynecologic oncologists about this rare disease. Case History. A 19-year-old female patient admitted to our hospital with the complaints of menstrual irregularity, hirsutism, and increased abdominal girth. Physical examination revealed bilateral ovarian mass, hirsutism, and ascites. Serum CA-125 levels were slightly elevated. Preoperative MRI study showed bilateral hypointense lobulated ovarian masses. With the initial diagnosis of ovarian tumor, we performed explorative laparotomy and excised both masses. Final pathology was reported as bilateral ovarian fibromatosis. Conclusion. Ovarian fibromatosis commonly presents with ascites and solid pelvic mass and can be misdiagnosed as a malignant ovarian tumor. In young patients, clinicians should consider ovarian fibromatosis in differential diagnosis of pelvic mass. Preoperative MRI findings and intraoperative frozen examination may be used to avoid unnecessary aggressive surgical management.

Ovarian carcinoma ascites spheroids adhere to extracellular matrix components and mesothelial cell monolayers

Objective. Ovarian carcinoma cells form multicellular aggregates, or spheroids, in the peritoneal cavity of patients with advanced disease. The current paradigm that ascites spheroids are non-adhesive leaves their contribution to ovarian carcinoma dissemination undefined. Here, spheroids obtained from ovarian carcinoma patients' ascites were characterized for their ability to adhere to molecules encountered in the peritoneal cavity, with the goal of establishing their potential to contribute to ovarian cancer spread. Methods. Spheroids were recovered from the ascites fluid of 11 patients with stage III or stage IV ovarian carcinoma. Adhesion assays to extracellular matrix (ECM) proteins and human mesothelial cell monolayers were performed for each of the ascites spheroid samples. Subsequently, inhibition assays were performed to identify the cell receptors involved. Results. Most ascites samples adhered moderately to fibronectin and type I collagen, with reduced adhesion to type IV collagen and laminin. Monoclonal antibodies against the β1 integrin subunit partially inhibited this adhesion. Ascites spheroids also adhered to hyaluronan. Additionally, spheroids adhered to live, but not fixed, human mesothelial cell monolayers, and this adhesion was partially mediated by β1 integrins. Conclusions. The cellular content of the ascites fluid has often been considered non-adhesive, but our findings are the first to suggest that patient-derived ascites spheroids can adhere to mesothelial extracellular matrix via β1 integrins, indicating that spheroids should not be ignored in the dissemination of ovarian cancer.

MR imaging of ovarian hemorrhage

Background: To review MR appearances of ovarian hemorrhage, and to describe its characteristic imaging findings. Methods: 12 women (age range, 20–44, mean, 26 years) with suspected ovarian hemorrhage underwent pelvic MR examinations. We retrospectively reviewed MR findings regarding signal intensities, localization, and wall enhancement of adnexal masses, and signal intensities of ascites. Results: Adnexal masses were detected in all cases. In eight cases, adnexal mass exhibited intermediate signal intensity on T1WI, and intermediate to low signal intensity on T2WI. In other case, adnexal mass exhibited marked hyperintensity on T1WI. In the remaining three cases, cystic mass with low signal intensity on T1WI and high signal intensity on T2WI was noted. Ascites was present in all cases, and showed intermediate signal on T1WI and intermediate to low signal on T2WI. Conclusions: In ovarian hemorrhage, hemorrhagic ascites and adnexal mass was visualized with specific MR signal intensity. Due to its sensitivity for identifying blood, MR imaging is useful in the diagnosis of ovarian hemorrhage, especially when ultrasonography findings are not definitive.

Effects of pentoxifylline in the prevention of ovarian hyperstimulation syndrome in a rabbit model

Tumor necrosis factor α (TNF-α) and other cytokines have been implicated in the pathogenesis of ovarian hyperstimulation syndrome (OHSS). Pentoxifylline, a methylxanthine derivative, was found to inhibit TNF-α synthesis. The aim of this study was to evaluate whether the use of petoxifylline would prevent the occurrence of OHSS in a rabbit model. Thirteen rabbits were divided into two groups. The first group (n = 6) were given pentoxifylline 15 mg/kg intravenously and the second group (n = 7) were given physiological serum 15 mg/kg before ovulation induction. Ovarian hyperstimulation was induced in rabbits by 200 IU equine chorionic gonadotropin on day 1 and 100 IU human chorionic gonadotropin on day 3. Blood samples were analyzed for TNF-α on days 1, 3 and 5. All animals were autopsied on day 6 to evaluate the ovarian weight, ascites formation and histopathological changes. There was no difference between groups regarding weight gain, ascites formation and plasma TNF-α levels (p > 0.05). Ovarian weight and number of ovulations were significantly lower in the pentoxifylline group than the control group (p < 0.05). Pentoxifylline did not prevent ascites formation despite the observed decrease in ovarian weight and number of ovulations in OHSS in a rabbit model.

Effects of pentoxifylline in the prevention of ovarian hyperstimulation syndrome in a rabbit model

Tumor necrosis factor α (TNF-α) and other cytokines have been implicated in the pathogenesis of ovarian hyperstimulation syndrome (OHSS). Pentoxifylline, a methylxanthine derivative, was foundto inhibit TNF-α synthesis. The aim of this study was to evaluate whether the use of petoxifylline would prevent the occurrence of OHSS in a rabbit model. Thirteen rabbits were divided into two groups.The first group (n = 6) were given pentoxifylline 15 mg/kg intravenously and the second group (n = 7) were given physiological serum 15 mg/kg before ovulation induction. Ovarian hyperstimulationwas induced in rabbits by 200 IU equine chorionic gonadotropin on day 1 and 100 IU human chorionic gonadotropin on day 3. Blood samples were analyzed for TNF-α on days 1, 3 and 5. All animals wereautopsied on day 6 to evaluate the ovarian weight, ascites formation and histopathological changes. There was no difference between groups regarding weight gain, ascites formation and plasma TNF-αlevels (p > 0.05). Ovarian weight and number of ovulations were significantly lower in the pentoxifylline group than the control group (p < 0.05). Pentoxifylline did not prevent ascitesformation despite the observed decrease in ovarian weight and number of ovulations in OHSS in a rabbit model.

Aggressive outpatient treatment of ovarian hyperstimulation syndrome with ascites using transvaginal culdocentesis and intravenous albumin minimizes hospitalization.

To assess the effectiveness of outpatient treatment of Ovarian Hyperstimulation Syndrome associated with ascites. Forty-eight patients diagnosed with ovarian hyperstimulation and ascites from 2,246 consecutive in vitro fertilization cycles were retrospectively studied. Patients were treated with outpatient transvaginal culdocentesis and rehydration with intravenous crystalloids and albumin every 1-3 days until resolution of symptoms or hospitalization was required. Outcomes measured included incidences of hospitalization, pregnancy outcomes, cycle characteristics, and oocyte donors versus nondonors comparisons. No complications occurred from outpatient treatments, and 91.6% of patients avoided hospitalization. The pregnancy rate in patients undergoing transfer was 84.7%, and the spontaneous loss rate was 16%. Overall, the estradiol on day of hCG was 4331 pg/mL (range 2,211-8,167), ascites removed was 1910 cm3 (122-4,000), and number of outpatient treatments was 3.4 (1-14). Nondonors averaged more outpatient treatments than donors (3.97 vs. 1.85), but similar rates of hospitalization (3/35 vs. 1/13). Outpatient treatment consisting of culdocentesis, intravenous rehydration, and albumin minimized the need for hospitalization in hyperstimulated patients.

Software compensation improves the analysis of heterogeneous tumor samples stained for multiparameter DNA flow cytometry

Background: High concentrations of propidium iodide (PI), in combination with fluorescein isothiocyanate (FITC) and R-phycoerythrin (RPE) used for multiparameter DNA flow cytometry (FCM), cause spectral cross-talk into the green fluorescence channel (FL1). We have evaluated the use of post-acquisition software compensation (N-Color Compensation) in order to correct this spectral cross-talk caused by PI. Method: Cell mixtures were prepared consisting of keratin 8/18 FITC labeled, keratin 8/18 RPE labeled, and unlabeled MCF-7 breast carcinoma cells. DNA was stained with PI (100 μM). Post-acquisition software compensation was applied to correct the spectral cross-talk of PI fluorescence. Secondly, the distribution of the Ki-67 (FITC) protein during the cell cycle (PI) of SiHa cervical carcinoma cells (no software compensation) was compared to the Ki-67 expression pattern of SiHa cells, simultaneously stained for keratin 8 (RPE), after applying software compensation. Finally, software compensation was used to compare the relative levels of PCNA and p53 expression in two clinical ovarian cancer ascites specimens, stained for PCNA or p53 (FITC), keratin 8/18 (RPE), and DNA (PI), with a known p53 status (positive and negative, respectively). Results: The Ki-67 cell cycle-dependent pattern of a triply stained sample (Ki-67 (FITC), keratin 8 (RPE), and DNA (PI)) is restored after software compensation and the results are comparable to the Ki-67 distribution of a sample stained solely for Ki-67 and DNA. P53 expression could only be resolved after using software compensation in the p53 positive ovarian ascites (OA) sample. Conclusions: We conclude that software compensation is a robust and reliable post-acquisition method for the correction of RPE/PI spectral cross-talk, permitting better identification of weakly expressed proteins in heterogeneous clinical tumor samples stained for multiple cellular antigens and DNA using PI.

Meigs’ syndrome with an elevated CA 125 from benign Brenner tumors

BACKGROUND: Meigs’ syndrome refers to solid, benign ovarian tumors, ascites, hydrothorax, and resolution of these signs after surgery. Meigs’ syndrome with an elevated CA 125 secondary to benign Brenner tumors is exceedingly rare. CASE: A postmenopausal woman presented with a large pelvic mass, ascites, and a right pleural effusion. Serum CA 125 was 759 IU/mL. Ascitic fluid, pleural fluid, and fine needle aspiration of the mass were without evidence of malignancy. Exploratory laparotomy with total abdominal hysterectomy and bilateral salpingo-oophorectomy revealed benign Brenner tumors. Immunohistochemical staining for CA 125 showed immunoreactivity in the omentum only. Postoperatively, her signs and symptoms resolved completely and did not recur. CONCLUSION: Cytologic or histologic confirmation of malignancy is imperative in patients with a pelvic mass, ascites, hydrothorax, and elevated CA 125 before initiating chemotherapy.