Marek Gogacz

Peritoneal fluid leptin concentration in infertile patients

Leptin may play a role in the regulation of menstrual cycle acting either directly on ovaries or at the level of the hypothalamic-pituitary axis. Peritoneal fluid is a biologically active environment that influences ovarian function but, on the other hand, concentration of many substances in peritoneal fluid can reflect the functional status of ovaries. In our study, we estimated leptin concentration in peritoneal fluid in relation to serum leptin concentration in infertile patients. A study group consisted of 31 infertile patients that underwent laparoscopy. In 15 patients, laparoscopy did not solve the problem and they were grouped as unexplained infertility, in eight patients we found endometriosis and eight patients were diagnosed as polycystic ovary syndrome (PCOS). We found significantly higher peritoneal fluid leptin concentrations in patients with unexplained infertility and endometriosis compared to those with PCOS. The plasma leptin concentration did not differ between the groups studied. There was no correlation between peritoneal fluid and plasma leptin concentrations in any patient group. Taking into account our results, the role of leptin in reproduction should be emphasized but further investigation is needed to determine its mechanism of action.

Increased percentage of Th17 cells in peritoneal fluid is associated with severity of endometriosis

AIM Th17 cells are a newly discovered T helper lymphocyte subpopulation, producing interleukin IL-17. Th17 cells are present in blood and peritoneal fluid (PF) at different stages of endometriosis. We aim to establish their potential importance in the pathogenesis and clinical features of the disease. METHODS The percentage of Th17 cells among T helper lymphocytes was determined in the PF and peripheral blood (PB) of patients with endometriosis and in the control group by flow cytometry using monoclonal antibodies: anti-CD-4-FITC, anti-CD-3-PE/Cy5, and anti-IL-17A-PE. RESULTS Th17 percentage is increased in PF in comparison with PB in both endometriotic patients and in the control group. In severe endometriosis, the percentage of Th17 cells in PF was higher than with early (I/II stage) endometriosis. A positive correlation between the percentage of Th17 cells in PF and the white blood cell count in PB was found in patients with endometriosis. CONCLUSION Targeting the activity of PF Th17 cells may have an influence on the proliferation of ectopic tissue and clinical manifestations of the disease.

Regulatory T lymphocytes and transforming growth factor beta in epithelial ovarian tumors-prognostic significance.

BackgroundRegulatory T lymphocytes (Treg) are characterized by the presence of CD4+ surface antigen. Today the transcription factor FOXP3 is considered to be the most specific marker of Treg cells. The aim of the study was to estimate the percentage of Treg in peripheral blood and the tissue of the epithelial ovarian tumor and blood serum TGF-beta concentrations and relationships between them. Moreover, the aim of the study was to answer the question whether the percentage of Treg lymphocytes affects the time of survival in patients with ovarian cancer.MethodsThe patients were divided into four groups, depending on the histopathological examination result: I – a group without any pathology within the ovaries (C; n = 20), II – a group with benign tumors (B; n = 25), III – with borderline tumors (BR; n = 11), IV – a group with cancer of the ovary (M; n = 24). The percentage of Treg lymphocytes in peripheral blood and the tissue was assessed using the flow cytometry method. TGF-beta cytokine concentration was estimated with the ELISA immunoenzymatic test. Statistical analysis of the results was conducted using the computer program Statistica 10.0PL (StatSoft, Inc).ResultsNo significant differences were found in percentages of Treg lymphocytes in peripheral blood between individual groups of patients (p = 0.11). However, we observed marked differences in the tissue of malignant and non-malignant tumors between individual groups of patients (p = 0.003). The analysis with the post hoc test revealed significantly higher TGF-beta concentration in the group of women with malignant tumors. Moreover, no relationship was found between TGF-beta concentration and the percentage of Treg cells in peripheral blood and tumors of the ovary. No correlation was found between the percentage of Treg lymphocytes in peripheral blood (p = 0.4) and the tissue of ovarian tumors (p = 0.3) and the time of survival of patients with ovarian cancer.ConclusionsThe recruitment of Treg lymphocytes toward the tumor is one of the mechanisms of escape of neoplasm from the response of the immune system. The percentage of Treg lymphocytes in peripheral blood and the neoplastic tissue does not influence the time of survival of patients with ovarian cancer.

Evaluation of Myeloid and Lymphoid Dendritic Cells in Peritoneal Fluid in Women with Non‐malignant Ovarian Tumors

Problem: Identification of myeloid and lymphoid dendritic cells (DCs) in peritoneal fluid (PF) and peripheral blood (PB) of patients with ovarian pathology.

Hyperprolactinaemia – a problem in patients from the reproductive period to the menopause

Hyperprolactinaemia especially affects women in reproductive age (90/100,000) but also often is diagnosed in menopause age and leads to disturbances in functioning of LH-RH neurons and, as a consequence, to a decrease of FSH and LH, which causes inhibition of oestradiol production. Prolactin is a peptide hormone, phylogenetically one of the oldest, stimulating cells of various organs, which is produced and secreted mainly by lactotrophic acidophilic cells of the anterior lobe of the pituitary. It influences the increase in the mass of the mammary glands, and stimulation and maintenance of lactation after delivery. There are a number of factors apart of pregnancy, delivery, and lactation than can influence secretion of the hormone in other physiological and pathological circumstances, like high-protein diet, stress, REM sleep, or neoplastic tumours, inflammatory diseases, chronic systematic diseases, thyroid hormonal changes, and drug intake. The purpose of this review is to summarise the current knowledge regarding the proper diagnosis and possible influence of hyperprolactinaemia on fertility and menopause symptoms and current treatment methods

Fas-Related Apoptosis of Peritoneal Fluid Macrophages in Endometriosis Patients: Understanding the Disease

Recent studies of the peritoneal cavity environment in endometriosis demonstrate quantitative and qualitative changes in the cells responsible for cell-mediated immunity. Such changes may have led to disturbances in the surveillance, recognition, and destruction of misplaced endometrial cells and might have, in fact, brought about the disease. The aim of the study was to assess CD95 (Fas) expression on (activated) peritoneal fluid (PF) macrophages, as well as to ascertain soluble Fas (sFas) concentration in the PF of endometriosis patients, as compared to the nonendometriotic group. The concentration of leukocytes in the PF, the percentage of cells expressing CD45+/CD14+, and the percentage of PF macrophages expressing the HLA-DR antigen were significantly higher in patients with stages I and II endometriosis. The percentage of Fas- (CD95+-) expressing macrophages was significantly higher in all stages of the disease, in comparison with controls. Moreover, the concentration of sFas in the PF of patients with moderate and severe endometriosis was significantly higher, as compared to the reference group. The high number of immune cells in PF in early stage endometriosis and their increased susceptibility to apoptosis confirm the role of the impaired peritoneal environment and immune defects in the development and progression of the disease.

Pregnancy complicated by ovarian planoepithelial carcinoma arising in mature cystic teratoma

An adnexal tumour can be expected in 2–10% of pregnancies. A mature cystic teratoma (MCT) constitutes 10–20% of all ovarian tumours and is the most frequent germ cell tumour of the ovary. The incidence of a malignant transformation in an MCT appears in 0.17–2% cases. The most frequent malignancy in an MCT is a planoepithelial carcinoma (squamous cell carcinoma) – 80–90% of cases (Hackethal et al. 2008). The malignant transformation of an MCT to a planoepithelial carcinoma in pregnancy is very rare. We found only two cases in the literature (Mekaru et al. 2008; Budiman et al. 2010). Although the management of ovarian tumours during a pregnancy is similar to the treatment in the case of nonpregnant patients, far more new studies are needed to confirm the most optimal management, due to the absence of standard recommendations. Case reports provide significant data on the management of these cancers (Palmer et al. 2009; Aggarwal and Kehoe 2011). We report an extremely rare case of a maternal planoepithelial carcinoma (G2) in a dermoid cyst detected during a caesarean section.

The Putative Role of TP53 Alterations and p53 Expression in Borderline Ovarian Tumors - Correlation with Clinicopathological Features and Prognosis: A Mini-Review

Borderline ovarian tumors (BOTs) represent an independent group among ovarian malignancies, being diagnosed at clinical stage earlier than invasive ovarian carcinomas (OCs) and characterized by a rather favorable outcome after careful surgical management. Data published worldwide showed a substantial discordance of p53 expression in BOTs. The purpose of this work was to present the current status of knowledge on the significance of TP53 gene and p53 protein product alterations in BOTs. In general, higher p53 expression patterns were reported for ovarian malignancies compared to BOTs. Serous, mucinous, and endometrioid BOTs differ substantially in relation to p53 immunostaining, but data concerning the relationship between the proteins immunoreactivity and other clinico-pathological variables are scarce. Finally, reports published to date support the view that TP53 alterations may not be commonly associated with the borderline phenotype of ovarian tumors but they probably occur during the development of invasive OCs. In light of these uncertainties, the impact of TP53 alterations and p53 expression on overall survival in women affected by BOTs requires further multi-institutional studies in large cohorts of patients.

Pregnant women suffering from uterine fibroids

Abstract Introduction. Uterine fibroids are the most frequent benign tumors affecting sexual organs in women. It is estimated that they affect 20% of the female population, with the frequency in pregnant women ranging between 0.1-5%. In spite of the progress in the field of medicine, the actual cause of uterine fibroids has yet to be discovered. Aim. Analysis of the recent methods of dealing with uterine fibroids during pregnancy. Material and methods. A review of literature about dealing with pregnant, lying-in and parturient women suffering from uterine fibroids. Results. The research studies by Aydeniz, Vergani, Rice showed that cesarean sections are much more frequent in pregnant women with uterine fibroids than in control group (52.9% vs 27.9%; 23% vs 12%; 35.1% vs 21.5%). However, it was shown that the rate of cesarean sections was much higher in women with uterine fibroids located in the lower part of the uterus than in the fundus uteri (respectively 39% and 18%). Also, the rate increased when the diameter of the fibroid exceeded 5 cm, unlike in case of those smaller than 5 cm (respectively 35% and 17%). Conclusions. 1. The number of cesarean sections in women with uterine fibroids is higher than in control group. 2. The frequency of cesarean sections in pregnancies with uterine fibroids depends on their position and size. 3. There is no relationship between the number of complications and the amount of fibroids in pregnant women. 4. There is no agreement concerning the recommendations for removing the fibroid during cesarean section.

Umbilical cord blood as a source of nerve and stem cells

OBJECTIVES The aim of the study was to assess proliferative ability of the stem cells in the umbilical cord blood and their potential to differentiate in in vitro culture. MATERIAL AND METHODS The material consisted of 14 samples of umbilical cord blood collected from the umbilical cord vein. Mononuclear cells were isolated using the method of density gradient medium. Next, CD34 cells were isolated from the interphase with the use of the VarioMACS sorter and anti-CD34 antibodies. Long-term cultures were conducted on Iscoves modified Dulbecco medium (IMDM) with addition of granulocyte-macrophage colony stimulating factor (GM-CSF) and nerve growth factor (NGF). Qualitative identification was performed using the May-Grunwald-Giemsy staining method, taking photographs with a confocal microscope, and with the immunoenzymatic method. RESULTS In our research, CD34+ stem cells constituted 1.16% of the mononuclear cells, and after centrifugation in medium 0.37% of leukocytes in whole umbilical cord blood. Even after 60 days of culture without addition of the growth factors, CD34+ cells were present in the fraction of adherent cells. After stimulation with GM-CSF and NGF a part of the umbilical cord blood cells were transformed into nerve cells (presence of neuron-specific enolase was shown) and into cells morphologically similar to fibroblast and dendritic cells. CONCLUSIONS After stimulation with GM-CSF and NGF cytokines, the umbilical cord blood cells proliferate in long-term medium, transform into nerve cells and into cells similar to fibroblast and dendritic cells.