ABSTRACT: To determine whether leiomyoma, adenomyosis and endometrial polyps are associated with changes in uterine cavity matrix metalloproteinases (MMP-2 and MMP-9) and cytokines.
Uterine cavity irrigation was performed in women with leiomyoma, adenomyosis and endometrial polyps, and in women with a normal uterus. MMP-2 and MMP-9 were assayed in the uterine washings by gelatin zymography. For individual subjects, the total MMP level was obtained by adding the semi-quantitative scores of band densities related to gelatinases in the zymograms. Interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and transforming growth factor-beta1 (TGF-beta1) were measured using enzyme-linked immunosorbant assay (ELISA) kits.
The uterine cavity of patients with leiomyoma, adenomyosis and endometrial polyps had significantly higher MMP scores than controls. Although the mean IL-1beta levels were elevated in uteri harboring a pathology compared with the normal uteri, the cytokine was significantly elevated only in the adenomyotic group. Significantly elevated levels of IFN-gamma were found in uteri with leiomyoma and endometrial polyps. Uterine washings from leiomyoma and adenomyosis contained significantly elevated mean levels of TGF-beta1 compared with controls, while TNF-alpha was significantly higher only in leiomyoma. When uterine cytokine levels were compared in relation to individual MMP levels a significant relationship was found between TGF-beta1 and elevated levels of MMP-9 and total MMPs in leiomyoma. A significant relationship was also found between IL-1beta and elevated levels of MMP-2, MMP-9 and total MMPs in the endometrial polyp group.
The uterine cavity in leiomyoma, adenomyosis and endometrial polyps contains elevated levels of MMPs and cytokines compared with the normal uterus. In some pathologies elevated cytokines are associated with elevated MMPs.
European Journal of Obstetrics & Gynecology and Reproductive Biology 01/2004; 111(2):197-203. · 1.97 Impact Factor
ABSTRACT: The known overexpression of matrix metalloproteinases (MMPs) by various tumors prompted a study to determine whether endometrial cancer could be detected by measuring MMPs in uterine cavity washings.
The study populations comprised 95 women being treated for endometrial cancer and 98 women with other gynecological conditions. A simple method was developed for performing uterine lavage and preparing cell-free uterine supernatants for MMP analysis. Gelatin zymography revealed elevated levels of latent and active forms of MMP-2 and MMP-9 in patients with endometrial cancer. For each patient individual bands of gelatinase activity were scored from 0 to 5 and summed to provide a total MMP score for analysis.
The mean MMP score in uterine washings of patients with endometrial cancer was 10.0 (range 1 to 22) compared with 0.8 (range 0 to 15) in the group without this cancer (P < 0.001). Receiver operating characteristic analysis showed that an MMP cutoff score of 3 gave a sensitivity of 98% and specificity of 91% for detecting endometrial cancer. With this MMP cutoff, a positive result was 11 times as likely in endometrial cancer compared to other conditions. The mean MMP score in the group with nodal metastases (14.1) was significantly higher than that without nodal involvement (9.4, P = 0.005). MMP-9 but not MMP-2 was significantly associated with nodal metastasis (P = 0.01). There was no significant association between MMP score and histological grade of tumor, vascular invasion, or depth of myometrial invasion.
Gelatinase measurement in uterine washings was reliable for confirming the presence of endometrial cancer in the population studied.
Gynecologic Oncology 08/2003; 90(2):318-24. · 3.89 Impact Factor
ABSTRACT: Cryopreservation of embryos and oocytes has become an essential service for infertility treatment. The clinical application of this technology should ensure optimal survival of the embryos and oocytes that are stored and subsequently thawed for transfer. The aim of this review is to compare the widely employed slow cooling procedures with vitrification to evaluate and recommend the more effective and safer procedure.
The review is mainly based on a comparison of the principles, procedures, and results reported in the literature. A historical description of vitrification and personal experiences with this technology are also included.
University-based hospitals and private clinics that treat infertility and have published information on cryopreservation.
Women being treated for infertility and reproductive technology clinics.
The application of slow cooling involving a range of cooling rates is compared with vitrification using rapid and ultrarapid cooling in simple containers. The purpose of both techniques is the induction of a glasslike state in cells to protect them from damage by ice crystals. The early development of vitrification involved the use of long pre-equilibration procedures. Improved methods resulted from the use of mixtures of penetrating and nonpenetrating solutes that are not toxic and a range of cooling rates.
Reported number of pregnancies established after transfer of embryos that were cryopreserved by vitrification, or transfer of embryos derived from vitrified oocytes.
Both slow cooling and vitrification procedures have resulted in the successful cryopreservation of human embryos and oocytes. Both procedures have resulted in healthy births, although the slow cooling of oocytes gives very low success rates. Vitrification is a promising novel technique in assisted reproductive technology, but comparative success rates are yet to be established.
Vitrification is a simple procedure that requires less time and is likely to become safer and more cost effective than slow cooling.
Fertility and Sterility 10/2002; 78(3):449-54. · 3.56 Impact Factor
Reviews in Endocrine and Metabolic Disorders 06/2002; 3(2):77-86. · 3.17 Impact Factor
ABSTRACT: The actions of the extracellular-matrix degrading enzymes, matrix metalloproteinases (MMPs), are implicated in tumorigenesis. The cellular localization of MMP-2, MMP-9, membrane type 1 (MT1)-MMP, tissue inhibitors of metalloproteinases (TIMPs) 1-3, and the presence of active gelatinases were investigated in endometrial carcinoma.
Endometrial carcinomas were grouped according to histologic grade (Grades 1-3), depth of myometrial invasion (0, < 50%, > 50%) and the presence of vascular/lymphatic invasion. Twenty-nine endometrial carcinoma biopsies were investigated immunohistochemically to determine the tissue localization of MMP-2 (gelatinase A), MMP-9 (gelatinase B), MT1-MMP, and TIMPs 1-3. In situ hybridization was performed to localize MMP-2 and MMP-9 mRNA. The presence of active gelatinases was assessed using in situ zymography.
Epithelial tumor cells were the main site of MMP-2, MMP-9, and MT1-MMP protein. Variable stromal cell localization was also observed, particularly in areas adjacent to tumor nests. Semiquantitative analysis revealed increases in MMP-9 and MMP-2 but not MT1-MMP staining scores in tumor epithelial cells in the transition from histologic Grade 1 to Grades 2 and 3. Matrix metalloproteinase-9 and MT1-MMP staining scores in tumor cells were significantly associated with the presence of myometrial invasion and vascular/lymphatic invasion, while MMP-2 did not correlate with these factors. In addition, MT1-MMP was co-localized with MMP-2, supporting its role in the activation of proMMP-2. Tumor cells from all histologic grades stained intensely for TIMP-2 and TIMP-3 proteins, while variable stromal staining was observed. In Grade 1 carcinomas TIMP-1 was predominantly immunolocalized to the stromal compartment with variable tumor cell localization being observed in Grades 2 and 3 carcinomas. Matrix metalloproteinase-9 and MMP-2 mRNAs were predominantly observed in tumor epithelial cells as well as in the stroma to varying degrees. In situ zymography revealed active forms of gelatinases at the cellular surface and in association with tumor epithelial cells within endometrial carcinoma tissues.
These data suggest that increasing expression of MMPs and endometrial carcinoma progression are closely related. Active gelatinases are present in endometrial carcinoma, resulting in alterations to the microenvironment that promote tumor invasion and metastasis.
Cancer 04/2002; 94(5):1466-75. · 4.77 Impact Factor