Survivin expression at the mRNA level in tumors and the protein concentration in the serum and peritoneal fluid in patients with serous ovarian tumors.

OBJECTIVES: Ovarian cancer is one of the gynecological cancers that have the worst prognosis. The expression of the proteins from the IAP family (inhibitor of apoptosis protein), including survivin, is observed in many types of cancer. The aim of the study was to evaluate survivin at the mRNA level in tumors and the protein concentration in the serum and peritoneal fluid of patients with serous ovarian cancer in order to assess the relationship between the concentration of survivin and the histological subtypes of cancer. MATERIAL AND METHODS: The study group consisted of 55 women, including patients with serous ovarian cancer (n = 30, nine low-grade serous carcinoma LGSC, 21 high-grade serous carcinoma HGSC), serous cysts (n = 10) and the control group (n = 15). The concentration of protein in the peritoneal fluid and serum was assessed using ELISA tests. The expression of survivin gene BIRC5 in the tumors was assessed using the RT-qPCR method. RESULTS: The data that was obtained indicated that the concentration of survivin was higher in the serum of the women with serous ovarian cancer compared those that had benign tumors (p < 0.05) and the control group (p < 0.001). The survivin concentration was also higher in both the serum and peritoneal fluid in the HGSC group compared to the LGSC group (p < 0.001). The mRNA level was highest in the HGSC group, and there was a statistically significant difference compared to those in the benign tumor group and HGSC group ( p < 0.05). CONCLUSIONS: The observed changes prove that the expression level increases significantly in HGSC in both the protein and mRNA levels. Based on these findings, it can be assumed that assessing this parameter could be a useful additional indicator of the progression and differentiation of this type of cancer. However, this requires further research in a larger group of patients and possibly in other types of ovarian cancer.

Expression pattern of circadian rhythm-related genes and its potential relationship with miRNAs activity in endometrial cancer.

OBJECTIVES: The circadian clock is an autonomous oscillator that controls key aspects of cell physiology, including metabolism, transcriptional state, and cell signaling. Disturbances of circadian rhythms lead to disruption of cell and tissue homeostasis, which promotes carcinogenesis. The aim of the study was to determine the expression of circadian rhythm-related genes in endometrial cancer and to select miRNAs involved in the regulation of their expression. MATERIAL AND METHODS: 50 endometrial tissue samples were collected from patients who underwent hysterectomy: 40 diagnosed with endometrial cancer and 10 without cancer. Expression profile of circadian rhythm-related genes was evaluated using microarrays and validated with RT-qPCR. MicroRNA expression was assessed using microarrays. Then mirTAR tool was used to identify miRNAs involved in the expression regulation of circadian rhythm-related genes. RESULTS: CLOCK expression is disrupted in endometrial cancer, which may be due to miR-15b, miR-331-3p and miR-200a overexpression. Elevated NPAS2 and CSNK1D levels may be associated with miR-432 silencing. In addition, high miR-874 and miR-200a expression may be potentially responsible for the reduction of PER3 level. CONCLUSIONS: Change of CLOCK, CSNK1D, NPAS2 and PER3 expression may suggest that circadian rhythms are disrupted in endometrial cancer. A possible mechanism of the observed changes may be related to miRNAs activity.

Analysis of CXCL8 and its receptors CXCR1/CXCR2 at the mRNA level in neoplastic tissue, as well as in serum and peritoneal fluid in patients with ovarian cance.

Understanding the relationship between the coexistence of inflammatory and neoplastic processes in ovarian cancer, particularly those involving chemokines and their receptors, may help to elucidate the involvement of the studied parameters in tumor pathogenesis and could lead to improved clinical applications. Therefore, the present study aimed to analyze the levels of C­X­C motif chemokine ligand 8 (CXCL8), and its receptors C­X­C chemokine receptor (CXCR)1 and CXCR2, in the serum and peritoneal fluid of women with ovarian cancer, and to evaluate the association between the expression of these parameters in tumor tissue and patient characteristics, particularly the degree of histological differentiation. The study group included women with ovarian cancer diagnosed with serous cystadenocarcinoma International Federation of Gynecology and Obstetrics stage IIIc and a control group, which consisted of women who were diagnosed with a benign lesion (serous cystadenoma). The transcript levels of CXCL8, CXCR1 and CXCR2 were evaluated using reverse transcription­quantitative PCR (RT­qPCR). The quantitative analysis was carried out using the LightCycler® 480 System and GoTaq® 1­Step RT­qPCR System, according to the manufacturers' instructions. The concentration of CXCL8 in serum and peritoneal fluid was determined using a Human Interleukin­8 ELISA kit, and the concentrations of CXCR1 and CXCR2 were determined using the CLOUD­CLONE ELISA kit. Local and systemic disturbances in immune and inflammatory responses involving the CXCL8 chemokine and its receptors indicated the involvement of these studied parameters in the pathogenesis of ovarian cancer. Immunoregulation of the CXCL8­CXCR1 system may influence the course of the inflammatory process accompanying ovarian cancer development, which may result in the identification of novel clinical applications; however, further studies are required.

Coffee consumption during pregnancy - what the gynecologist should know? Review of the literature and clinical studies.

Coffee is one of the most consumed beverages in the world. The impact of coffee consumption on human health has been the subject of many clinical studies and meta-analyses. Taking into account the results of these studies, it can be concluded that coffee has a number of health benefits in terms of the population, including the reduction of the risk of death from any cause. From a clinical point of view, the safety of coffee consumption in a specific subpopulation of pregnant women is important. A large percentage of women continue to consume this drink during pregnancy, while a significant proportion of them exceed the permissible daily dose of caffeine (≤ 200 mg). During pregnancy, the metabolism of caffeine slows down significantly, which prolongs its action and penetrates into the body of the fetus. These biochemical observations have become the driving force behind numerous clinical studies assessing the impact of coffee consumption during pregnancy on its course, complications and the health of the newborn. This review article summarizes the current knowledge of these important issues.

Grading of endometrial cancer using 1H HR-MAS NMR-based metabolomics.

The tissue metabolomic characteristics associated with endometrial cancer (EC) at different grades were studied using high resolution (400 MHz) magic angle spinning (HR-MAS) proton spectroscopy. The metabolic profiles were obtained from 64 patients (14 with grade 1 (G1), 33 with grade 2 (G2) and 17 with grade 3 (G3) tumors) and compared with the profile acquired from 10 patients with the benign disorders. OPLS-DA revealed increased valine, isoleucine, leucine, hypotaurine, serine, lysine, ethanolamine, choline and decreased creatine, creatinine, glutathione, ascorbate, glutamate, phosphoethanolamine and scyllo-inositol in all EC grades in reference to the non-transformed tissue. The increased levels of taurine was additionally detected in the G1 and G2 tumors in comparison to the control tissue, while the elevated glycine, N-acetyl compound and lactate-in the G1 and G3 tumors. The metabolic features typical for the G1 tumors are the increased dimethyl sulfone, phosphocholine, and decreased glycerophosphocholine and glutamine levels, while the decreased myo-inositol level is characteristic for the G2 and G3 tumors. The elevated 3-hydroxybutyrate, alanine and betaine levels were observed in the G3 tumors. The differences between the grade G1 and G3 malignances were mainly related to the perturbations of phosphoethanolamine and phosphocholine biosynthesis, inositol, betaine, serine and glycine metabolism. The statistical significance of the OPLS-DA modeling was also verified by an univariate analysis. HR-MAS NMR based metabolomics provides an useful insight into the metabolic reprogramming in endometrial cancer.

The Assessment of IL-21 and IL-22 at the mRNA Level in Tumor Tissue and Protein Concentration in Serum and Peritoneal Fluid in Patients with Ovarian Cancer.

The aim of the analysis was for the first time to assess the expression of genes encoding IL-21 and IL-22 at the mRNA level in ovarian tumor specimens and the concentration of these parameters in serum and peritoneal fluid in patients with ovarian serous cancer. The levels of IL-21 and IL-22 transcripts were evaluated with the use of the real-time RT-qPCR. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of proteins. Quantitative analysis of IL-21 gene mRNA in the tumor tissue showed the highest activity in the G1 degree of histopathological differentiation and was higher in G1 compared to the control group. The concentration of IL-21 and IL-22 in the serum and in the peritoneal fluid of women with ovarian cancer varied depending on the degree of histopathological differentiation of the cancer and showed statistical variability compared to controls. The conducted studies have shown that the local and systemic changes in the immune system involving IL-21 and IL-22 indicate the participation of these parameters in the pathogenesis of ovarian cancer, and modulation in the IL-21/IL-22 system may prove useful in the development of new diagnostic and therapeutic strategies used in patients, which require further research.

Expression Profile of EMT-related Genes and miRNAs Involved in Signal Transduction via the Wnt Pathway and Cadherins in Endometrial Cancer.

BACKGROUND: Epithelial-Mesenchymal Transition (EMT) is a molecular reprogramming that leads to an increased ability to migrate, which can promote invasion and metastasis. EMT can be initiated in response to the activity of signaling pathways such as Wnt as well as miRNAs. OBJECTIVE: The aim of the study was to determine the expression profile of EMT-related genes involved in signal transduction via the Wnt pathway and cadherins and to assess which miRNAs can participate in the regulation of their expression. METHODS: The study material consisted of 50 endometrial samples: 40 with diagnosed endometrial cancer and 10 without neoplastic changes. The expression profile of EMT-related genes was assessed with microarrays and validated by RT-qPCR. MicroRNA expression profiling was performed using microarrays. It was also determined which miRNAs may participate in the expression regulation of EMT-related genes. RESULTS: CDH1 overexpression was observed in all three endometrial cancer grades using both mRNA microarrays and RT-qPCR. The microarray experiment showed a decrease in CDH2 level regardless of the endometrial cancer grade, however, it was only partially validated with RT-qPCR. Low levels of WNT2, WNT4, WNT5A have also been observed. Decreased expression of WNT2 and WNT5A may be caused by miR-331-3p and miR-200b-5p, respectively. CONCLUSION: The Wnt signaling is disrupted in endometrial cancer, which may be due to miR-331- 3p and miR-200b-5p activity. In addition, a change in WNT5A level in endometrial cancer compared to control may indicate that it acts as a suppressor gene and that its low expression is associated with tumor progression.

Leydig cell ovarian tumor - clinical case description and literature review.

Leydig cell ovarian tumors constitute not only a medical problem for clinicians but also a social problem - which is why women with symptoms of hirsutism relatively quickly contact physicians for medical consultation. Leydig cell ovarian tumor is a rare sex cord-gonadal stromal tumor which constitutes less than 0.5% of ovarian tumors. These cancers appear at all ages but the majority of the cases concern women in the perimenopause. In the majority of cases (70-85%), the growth is accompanied by androgen secretion, together with virilization and hirsutism. The presence of hormonally active ovarian cancers should be suspected in cases of rapidly growing symptoms of masculinization, especially when the level of free testosterone in the blood exceeds the upper limit for the given age more than three times. In diagnosing postmenopausal hyperandrogenism, it is necessary to take into account hormonally active ovarian tumors, as well as adrenal cancers. It is important to exclude other causes of hyperandrogenism, e.g. endocrinopathies (acromegaly or hypothyroidism), or iatrogenic and idiopathic factors. In order to make the diagnosis and implement the proper treatment method faster, an interdisciplinary team of physicians specializing in endocrinology, gynecology and oncology is crucial. This paper contains a study of two cases concerning Leydig cell ovarian tumors in women of postmenopausal age with symptoms of masculinization and hirsutism.

Basaloid squamous cell carcinoma of the uterine cervix coexisting with undifferentiated sarcoma.

Basaloid squamous cell carcinoma is uncommon tumor and has received little recognition, there are only a few cases in the medical literature. Correct diagnosis is important due to the high malignancy of this cancer and poor prognosis. We present a rare and interesting case of a basaloid squamous cell carcinoma of the uterine cervix in a 71-year-old female with prolapse of the uterus with co-existing atypical undifferentiated sarcoma.

Interplay between miRNAs and Genes Associated with Cell Proliferation in Endometrial Cancer.

Endometrial cancer develops as a result of abnormal cell growth associated with uncontrolled cell proliferation, excessive activation of signaling pathways and miRNA activity. The aim of this study was to determine the expression profile of genes associated with cell proliferation and to assess which miRNAs can participate in the regulation of their expression. The study enrolled 40 patients with endometrial cancer and 10 patients without neoplastic changes. The expression profile of genes associated with cell proliferation and the expression profile of miRNAs were assessed using microarrays. RT-qPCR was performed to validate mRNA microarray results. The mirTAR tool was used to identify miRNAs that regulate the activity of genes associated with cell proliferation. Decreased expression of IGF1 and MYLK, as well as SOD2 overexpression, were observed in endometrial cancer using both mRNA microarrays and RT-qPCR. Microarray analysis showed low levels of NES and PRKCA, but this was only partially validated using RT-qPCR. Reduced activity of MYLK may be caused by increased miR-200c, miR-155 and miR-200b expression. Cell proliferation is disturbed in endometrial cancer, which may be associated with an overexpression of miR-200a, miR-200c, and miR-155, making it a potential diagnostic marker.

CA125 as a Marker of Heart Failure in the Older Women: Population-Based Analysis.

(1) Background: Cancer antigen 125 (CA125) is a glycoprotein that is expressed by tissue derived from coelomic epithelium in the pleura, peritoneum, pericardium. It has been shown that CA125 concentrations are correlated with NT-proBNP in older with congestive heart failure (HF). We conducted a study on the association between concentrations of CA125 and NT-proBNP in a population-based cohort of older Polish women. (2) Methods: The current research is sub-study of a large, cross-sectional research project (PolSenior). The study group consisted of 1565 Caucasian women aged 65-102 years. To assess the relationship between CA125 and other variables a stepwise backward multivariate normal and skew-t regression analyses were performed. (3) Results: The median of CA125 concentration was 13.0 U/mL and values over the upper normal range limit (35 U/mL) were observed in 5.1% (n = 79) of the study cohort. The concentration of CA125 was positively related to age, hospitalization for HF and history of atrial fibrillation and chronic obstructive pulmonary disease, levels of NT-proBNP, IL-6, hs-CRP and triglycerides. We found, in multivariate analyses, that increased CA125 levels were independently associated with log10 (IL-6) (ß = 11.022), history of hospitalization for HF (ß = 4.619), log10 (NT-proBNP) (ß = 4.416) and age (ß = 3.93 for 10 years). (4) Conclusion: Despite the association between CA125 and NT-proBNP, the usefulness of CA125 for the detection of HF in older women is limited by factors such as inflammatory status and age.

Dietary supplementation usage by pregnant women in Silesia - population based study.

OBJECTIVES: Despite wide access to gynecological and obstetric advice, informational campaigns, and information online and in magazines aimed at pregnant women, there is a worryingly high percentage of women who still do not use recommended dietary supplementation. The aim of this study was to assess the frequency of micronutrient supplementation by pregnant women and to specify the determinants that impact decisions concerning supplementation. MATERIAL AND METHODS: A cross-sectional survey was conducted between June 2016 and May 2017 among a group of pregnant women visiting gynecological and obstetric clinics in the Silesia region, who have completed an authorized questionnaire developed for the purpose of this study. The questionnaire addressed the women's dietary habits, micronutrient supplementation use, as well as their socio-economic status. Completed questionnaires were obtained from 505 pregnant women. RESULTS: Microminerals and vitamins supplementation during pregnancy was declared by 410 (81.2%) women. The most often used supplement was folic acid (62%). More than one-third of pregnant women (38.4%) declared vitamin D intake. Among the recommended supplements, the least commonly used (30.3%) were polyunsaturated fatty acids (PUFA). Factors contributing to supplementation use during pregnancy are past history of miscarriage and socioeconomic factors, such as: place of residence, financial situation and level of education. Inhabitants of larger cities, women with better self-perceived financial situations, higher education levels and those presenting past history of miscarriage took the supplements significantly more often. CONCLUSIONS: Lower levels of education, low-income financial status and living in rural localities are among the factors correlating with worse adherence to supplementation guidelines.

The role of complement components C1q, MBL and C1 inhibitor in pathogenesis of endometriosis.

PURPOSE: The purpose of the work was to evaluate possible associations between the complement components C1q, mannose-binding lectin (MBL) and C1 inhibitor (C1INH) with pathogenesis of endometriosis. METHODS: Concentrations of C1q, MBL and C1INH were measured by ELISA in peritoneal fluid (PF) in 80 women with or without endometriosis. RESULTS: Significantly higher PF levels of C1q, MBL and C1INH in women with endometriosis compared to control group were observed (p < 0.0001). A higher concentration of the studied parameter was found in PF of women at the early stage of the disease, as compared to women with advanced endometriosis (p < 0.0001). CONCLUSIONS: Our research suggests that in the peritoneal cavity in women with endometriosis there are abnormal regulations of both the classical and lectin pathways of the complement system. This can suggest impairments in purification of peritoneal cavity from ectopic endometrial cells and augmented local inflammation in endometriosis patients.

Exenatide modulates tumor-endothelial cell interactions in human ovarian cancer cells.

Diabetes and cancer are prevalent diseases whose incidence is increasing globally. Diabetic women have a moderate risk increase in ovarian cancer, suggested to be due to an interaction between these two disorders. Furthermore, patients manifesting both diseases have associated worse prognosis, reduced survival and shorter relapse-free survival. According to current recommendations, incretin drugs such as Exenatide, a synthetic analog of Exendin-4, and Liraglutide are used as therapy for the type 2 diabetes (T2D). We studied the effects of GLP-1 and Exendin-4 on migration, apoptosis and metalloproteinase production in two human ovarian cancer cells (SKOV-3 and CAOV-3). Exendin-4 inhibited migration and promoted apoptosis through caspase 3/7 activation. Exendin-4 also modulated the expression of key metalloproteinases (MMP-2 and MMP-9) and their inhibitors (TIMP-1 and TIMP-2). Vascular endothelial cells, which contribute to the formation and progression of metastasis, were also analyzed. TNF-α-stimulated endothelial cells from iliac artery after Exendin-4 treatment showed reduced production of adhesion molecules (ICAM-1 and VCAM-1). Additionally, incretin treatment inhibited activation of apoptosis in TNF-α-stimulated endothelial cells. In the same experiment, MMPs (MMP-1 and MMP-9), which are relevant for tumor development, were also reduced. Our study demonstrated that incretin drugs may reduce cancer cell proliferation and dissemination potential, hence limiting the risk of metastasis in epithelial ovarian cancer.

Bronchial hyperreactivity in perimenstrual asthma is associated with increased Th-2 response in lower airways.

BACKGROUND: Perimenstrual asthma (PMA) is a commonly observed, usually difficult-to-treat asthma phenotype. The mechanisms underlying this phenomenon remain unexplained. The aim of the study was to assess the degree of airway hyperresponsiveness and its relationship to proinflammatory cytokines concentration in lower airways of PMA compared to non-PMA patients. METHODS: Premenopausal women with regular menstrual cycles diagnosed as: PMA (n=12), non-PMA asthmatics (n=9), and healthy controls (n=10) were prospectively followed for 10 weeks over two consecutive menstrual cycles. The bronchial responsiveness (BR) test to methacholine was performed in each subject prior to the study. The serum for total immunoglobulin E (IgE) concentrations was taken and sputum was induced in the 26th day of each of the two cycles. Sputum concentration of eotaxin, IL-4 and IL-10 were measured by ELISA. RESULTS: Levels of BR to metacholine as well, as total blood IgE concentrations in PMA subjects were significantly higher than in non-PMA asthmatics and healthy controls (P=0.001, P=0.022 respectively) and correlated with each other (P=0.030; r =-0.65). Sputum eotaxin and IL-4 concentrations in luteal phase were increased in PMA patients when compared with non-PMA asthmatics (P=0.016; P=0.041, respectively) and healthy subjects (P<0.001 both cytokines). No differences for the sputum levels of IL-10 among studied groups were seen. CONCLUSIONS: BR level in perimenstrual asthma is higher than in non-PMA asthmatics and correlates with increased total IgE serum concentration. The increased level of BR in PMA patients is associated with a shift in the type-1/type-2 cytokine balance toward a type-2 response.

Identification of a gene expression profile associated with the regulation of angiogenesis in endometrial cancer.

The publication of the human genome sequence provided direction in the search for novel diagnostic and therapeutic methods for the treatment of human diseases. The aim of the present study was to investigate the hypothesis that the expression profile of genes involved in the regulation of angiogenesis may be a marker in endometrial cancer that facilitates the diagnosis and prognosis of patients, as well as the identification of novel therapeutic targets. The current study included 36 patients with grade (G) 1 to 3 endometrial cancer, and a control group of patients consisting of females that qualified for the removal of the uterus. Out of these, 28 samples (control, 3; G1, 7; G2, 12; and G3, 6) were selected for microarray analysis. Molecular analysis of the endometrial samples involved the extraction of total RNA, purification of the obtained extracts and subsequent analysis of the gene expression profiles using an oligonucleotide microarray technique (GeneChip® Human Genome U133A plates). The results indicated that the mRNA expression profile of genes involved in the regulation of angiogenesis varies depending on the degree of histological differentiation of endometrial adenocarcinoma. Similar results were obtained from descriptive statistics characterizing the expression profile of 691 mRNAs associated with the regulation of angiogenesis in the groups of patients with endometrial adenocarcinoma. In addition, the results of the present study indicated that neuropilin2 (NRP2) may serve an important role in the activity of endothelial cells, and may affect vascular endothelial growth factor, and potentially plexins and integrins via regulation of their functions. An understanding of how these proteins interact remains to be determined; however, elucidating these interactions may provide an explanation for the mechanisms underlying angiogenesis. In conclusion, the results of the present study suggest that NRP2 may be a valuable target for investigation in future pharmacological studies involving angiogenesis in endometrial cancer.

Cesarean scar pregnancy - case report and review.

Implantation of the embryo in the cesarean section scar is a rare form of ectopic pregnancy. Such condition poses significant threat to a woman's health and live, therefore, requires accurate diagnosis and rapid implementation of treatment. The following article presents the case of a patient with a pregnancy located in the scar after cesarean section treated in the Department of Gynecology and Obstetrics, Medical University of Silesia in Katowice, Faculty of Medicine in Katowice.

Clinical expression of Holt-Oram syndrome on the basis of own clinical experience considering prenatal diagnosis.

OBJECTIVES: Holt-Oram syndrome manifests with defects of upper limbs, pectoral girdle and cardiovascular system. The aim of this paper was to present complex clinical picture of the syndrome and its variable expression on the example of the family diagnosed genetically on the neonatal ward, after proband's prenatal examination. MARETIAL AND METHODS: Nine family members were tested for TBX5 gene mutation. RESULTS: Four of family members were diagnosed with Holt-Oram syndrome and five had correct genetic test results. The diagnosis allowed to identify a genetic risk family and enabled to provide them with genetic counselling. CONCLUSIONS: Diagnosis of Holt-Oram syndrome is possible as early as in prenatal period and it can be verified by genetic tests.

Changes in expression of cartilaginous genes during chondrogenesis of Wharton's jelly mesenchymal stem cells on three-dimensional biodegradable poly(L-lactide-co-glycolide) scaffolds.

BACKGROUND: In cartilage tissue regeneration, it is important to develop biodegradable scaffolds that provide a structural and logistic template for three-dimensional cultures of chondrocytes. In this study, we evaluated changes in expression of cartilaginous genes during in vitro chondrogenic differentiation of WJ-MSCs on PLGA scaffolds. METHODS: The biocompatibility of the PLGA material was investigated using WJ-MSCs by direct and indirect contact methods according to the ISO 10993-5 standard. PLGA scaffolds were fabricated by the solvent casting/salt-leaching technique. We analyzed expression of chondrogenic genes of WJ-MSCs after a 21-day culture. RESULTS: The results showed the biocompatibility of PLGA and confirmed the usefulness of PLGA as material for fabrication of 3D scaffolds that can be applied for WJ-MSC culture. The in vitro penetration and colonization of the scaffolds by WJ-MSCs were assessed by confocal microscopy. The increase in cell number demonstrated that scaffolds made of PLGA copolymers enabled WJ-MSC proliferation. The obtained data showed that as a result of chondrogenesis of WJ-MSCs on the PLGA scaffold the expression of the key markers collagen type II and aggrecan was increased. CONCLUSIONS: The observed changes in transcriptional activity of cartilaginous genes suggest that the PLGA scaffolds may be applied for WJ-MSC differentiation. This primary study suggests that chondrogenic capacity of WJ-MSCs cultured on the PLGA scaffolds can be useful for cell therapy of cartilage.