Cervical mucus proteome in endometriosis.

BACKGROUND: Endometriosis is a chronic gynecological inflammatory disease characterized by the presence of functional endometrial glands and stroma outside of the uterine cavity. It affects 7-10% of women of reproductive age and up to 50% of women with infertility. The current gold standard for the diagnosis combines laparoscopic evaluation and biopsy of the visualized lesions. However, laparoscopy requires general anesthesia and developed surgical skills and it has a high procedural cost. In addition, it is associated with the risk, although rare, of potential intraoperative or postoperative complications. To date, several noninvasive biomarkers have been proposed; however, no definite diagnostic biomarker is yet available. The aim of this study was to characterize the CM proteome in patients with endometriosis using high resolution mass spectrometry-based proteomics, implemented by bioinformatic tools for quantitative analysis, in order to investigate the pathophysiological mechanisms of endometriosis. METHODS: Cervical mucus samples were collected from patients affected by endometriosis and fertile controls. An aliquot of the soluble acidic fraction of each cervical mucus sample, corresponding to 0.5 mg of total protein, was left to digest with sequencing grade modified porcine trypsin. The peptides were analyzed by LC-MS/MS on a high resolution Orbitrap Elite mass spectrometer and data were evaluated using bioinformatic tools. RESULTS: We aimed at the first total profiling of the cervical mucus proteome in endometriosis. From the list of identified proteins, we detected a number of differentially expressed proteins, including some functionally significant proteins. Six proteins were quantitatively increased in endometriosis, almost all being involved in the inflammatory pattern. Nine proteins were quantitatively reduced in endometriosis, including some proteins related with local innate immunity (CRISP-3 and Pglyrp1) and protection against oxidative stress (HSPB1). Fifteen proteins were not detected in endometriosis samples including certain proteins involved in antimicrobial activity (SLURP1 and KLK13) and related to seminal plasma liquefaction and male fertility (KLK13). CONCLUSIONS: This is the first application of high resolution mass spectrometry-based proteomics aimed in detecting an array of proteins in CM to be proposed for the noninvasive diagnosis of endometriosis. This chronic disease presents in CM an inflammatory protein pattern.

The role of sonographic examination in the follow-up of gynecological neoplasms.

OBJECTIVE: The role and type of procedures of follow-up in patients with gynecological tumors are still a debatable issue. We prospectively analyzed the role of routine transabdominal and transvaginal ultrasound examination (US) in the detection of recurrent disease in gynecologic cancer patients. METHODS: Among 552 patients who underwent surgery for gynecological cancer, 385 were available for the analysis. Follow-up examinations included clinical examination, serum tumor marker assay, transvaginal and transabdominal sonography and CT scan/MRI. RESULTS: Positive US examination was documented in 83/385 patients (21.5%). In the overall series, the positive predictive value (PPV) of US examination was 100%, while the negative predictive value (NPV) was 92.7% failing to identify 22 cases of recurrences. When considering the subgroup of patients with positive clinical examination or abnormal tumor marker, positive US analysis was able to identify 66/66 cases of recurrence (PPV=100%), but exhibited an NPV of 22.2%, with 21/27 (77.8%) false negative cases. Conversely, in cases without clinical/serological signs of disease, positive US recognized all cases of recurrences (17/17, PPV=100%), and exhibited a very high NPV, with only 1 false negative case out of 275 (0.4%). The US detected recurrences appeared as a solitary lesion in 38/75 (50.6%) patients and in 28/75 (37%) appeared located centrally in the pelvis. The sonographic pattern of the ovarian recurrences was a solid lesion in most (33/40, 82%) cases. In the other gynecological tumors, the lesions appeared as solid masses in 33/35 (94%) cases. CONCLUSIONS: We showed that routine US might play a role in the follow-up of gynecologic malignancies, especially in the group of asymptomatic patients, while CT/MRI imaging might be more properly applied to patients with clinical or serological signs of disease.

The use of contrasted transvaginal sonography in the diagnosis of gynecologic diseases: a preliminary study.

OBJECTIVE: The purpose of this study was to evaluate the efficacy of a new contrast-dedicated ultrasound technology, contrast-tuned imaging (CnTI), implemented on an endovaginal probe and using the second-generation contrast agent SonoVue (Bracco International BV, Amsterdam, the Netherlands), compared with the standard ultrasound examination in different gynecologic diseases. METHODS: Eighty-nine patients were enrolled in the study in 4 different clinical centers. The study included 40 patients with uncertain pelvic adnexal masses, 10 patients with pelvic masses indicative of recurrences of gynecologic tumors, 26 patients with uterine pathologic features, and 13 patients with cervical lesions. RESULTS: Application of CnTI technology after the SonoVue injection gave a picture of the intralesional microvascularization dramatically different from that obtained during color Doppler examination. Of the 40 pelvic masses, 15 (37.5%) were considered benign and 25 (62.5%) were considered malignant at B-mode and color Doppler examinations. Contrast-enhanced sonography showed no intralesional contrast perfusion in 11 (73%) of 15 cases, and all these were benign at final diagnosis. Of the 4 (27%) cases that had perfusion, 2 were malignant. Conversely, of the 25 cases with positive findings at color Doppler examination and therefore expected to show the appearance of contrast tissue-filling morphologic characteristics, 13 (52%) were malignant at final diagnosis. For evaluation of uterine pathologic features, the CnTI-SonoVue technology did not appear to be superior to the B-mode and color Doppler examinations; however, for the evaluation of cervical cancer, CnTI-SonoVue technology revealed a better definition of the margins of the neoplastic lesions in 4 (40%) of 10 cases. CONCLUSIONS: In the evaluation of uncertain pelvic masses, the CnTI technology led to an improvement in the ability of the practitioner to differentiate benign from malignant adnexal lesions.

Angiographic power 3-dimensional quantitative analysis in gynecologic solid tumors: feasibility and reproducibility.

OBJECTIVE: To determine the reproducibility of a simplified method of power Doppler 3-dimensional (3D) sonographic examination. METHODS: Twenty-nine patients with solid pelvic masses underwent transvaginal 3D power Doppler evaluation. The volume of interest was obtained by drawing the margins of the largest section of the mass in the 3 orthogonal planes. The 3D vascular parameters ("relative color," "average color," and "flow measure") obtained by our method were compared with those calculated by a manufacturer-suggested model based on several parallel section planes drawn on the longitudinal frames. The intraobserver variability was quantified on 5 different 3D images acquired by the same operator at 5-minute intervals for each patient. The intraobserver variability was also assayed in 10 patients at 24-hour intervals. Ten patients were scanned by a second sonographer for interobserver variability. RESULTS: There was high agreement between the 3D parameters obtained with the 2 methods. The 3D indices were similar in repeated observations at 5-minute intervals (median coefficients of variation for relative color, average color, and flow measure, 10.9, 4.5, and 13.0, respectively) and at 24-hour intervals (intraclass correlation coefficients for relative color, average color, and flow measure, 0.920, 0.978, and 0.978) and by the second sonographer (interclass correlation coefficients for relative color, average color, and flow measure, 0.978, 0.966, and 0.997). CONCLUSIONS: The acceptable rates of intraobserver and interobserver variability make this approach potentially suitable for research protocols.

Gemcitabine: current role and future options in the treatment of ovarian cancer.

Epithelial ovarian cancer is the gynecological malignancy with the highest mortality. The standard therapeutic approach for patients with advanced-stage epithelial ovarian cancer has been cytoreductive surgery followed by combination chemotherapy. Despite improvements in outcome via carboplatin/paclitaxel based chemotherapy, 30% of patients with ovarian cancer fail to respond to primary therapy; moreover, 55-75% of responders relapse within 1 or 2 years from the end of primary treatment and die of the disease within 5 years from their initial diagnosis. Gemcitabine has been shown to be active as a single agent and in combination with other drugs, including carboplatin and paclitaxel, in the treatment of patients with recurrent ovarian cancer. It is currently under evaluation in new combinations for initial therapy in ovarian cancer patients.

Dietary soy modulation of biochemical parameters in DMBA-induced mammary tumors.

The aim of this study was to extend our previous observations on the soy modulation of biochemical parameters in 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumors, by simultaneously investigating the expression of estrogen receptor-alpha (ERalpha), estrogen receptor-beta (ERbeta), progesterone receptor (PR), apoptosis, neu, and markers of cell proliferation, such as proliferating cell nuclear antigen (PCNA) by immunohistochemistry. The percentage of ERalpha positive tumors was 65.8% in masses from control animals, and significantly dropped to 36.8% in tumors from soy treated rats (P=0.010). The percentage of ERbeta positive tumors was 70.3% in masses from control animals vs. 50.0% in tumors from soy exposed animals (P=0.066). Moreover, the percentage of cases which were both ERalpha and ERbeta positive was significantly lower (17.6%) in soy treated than in control animals (51.3%) (P=0.006). The percentage of PR positive tumors was 34.2% in control animals vs. 2.6% in tumors from soy treated rats (P=0.0006). There were no statistically significant differences in the percentage of tumors positively stained for neu, apoptosis, or PCNA, in control vs. soy treated rats. However, when analyzing the reciprocal correlation among the different biochemical parameters we showed that, in treated animals, the majority of ERalpha positive tumors (91.7%) were also PCNA positive (P=0.036). The median percentage of PCNA positivity was significantly higher in ERalpha positive than in ERalpha negative tumors (25 vs. 5%) (P=0.0031). Moreover, an association was found between PCNA and neu status since all neu positive tumors were also PCNA positive (P=0.011).