Human amniotic fluid as a source of stem cells.

Human amniotic fluid collected during amniocentesis contains a heterogeneous population of differentiated and undifferentiated cells. Properties and number of these cells vary depending on the gestational age and the presence of potential fetal pathologies. The aim of this study was to analyze the effects of maternal, fetal, and environmental factors on the success rates of amniotic fluid stem cell cultures, the number of human amniotic fluid stem cells (hAFSC), their growth rates in primary cultures, and the number of cell passages. The study included 355 patients qualified for genetic amniocentesis at the Prenatal Genetic Unit, Department of Obstetrics, Gynecology and Oncologic Gynecology, Nicolaus Copernicus University Medical College in Bydgoszcz in 2011-2017. The mean age of the study participants was 34 ± 6.2 years, and mean gravidity amounted to 2.48 ± 1.4. Amniotic fluid sample volume turned out to be a highly significant (p < 0.01) predictor of culture success, and the relationship was particularly evident in women older than 40 years. Another highly significant predictor of culture success was the presence of two cell populations in the sample (p < 0.01). The likelihood of culture success correlated significantly (p < 0.05) with the season of the year at the time of amniocentesis. The number of cell passages differed significantly depending on the maternal age (p < 0.01). The number of passages also showed a highly significant relationship with the season of the year the sample was obtained (p < 0.01). Younger maternal age was identified as a determinant of high passage number (≥3), and another highly significant determinant of high passage number was the presence of two cell populations in the amniotic fluid sample (p < 0.01). Percentage of successfully established hAFSC cultures and the number of passages depended on amniotic fluid volume, the presence of two cell populations within the sample, and the season of the year. Individual characteristics of the donors, such as age and gravidity, did not exert a significant effect on the number of isolated hAFSCs and the rate of their growth. Patients' place of residence, fetal karyotype, transportation time, and purity of the samples did not affect the success rates for primary cultures and the number of passages.

Assessment of the SFlt-1 and sFlt-1/25(OH)D Ratio as a Diagnostic Tool in Gestational Hypertension (GH), Preeclampsia (PE), and Gestational Diabetes Mellitus (GDM).

BACKGROUND: Placental soluble fms-like tyrosine kinase-1 (sFlt-1), an antagonist of vascular endothelial growth factor, is considered an etiological factor of endothelial damage in pregnancy pathologies. An increase in the sFlt-1 level is associated with alterations of endothelial integrity. In contrast, vitamin D exerts a protective effect and low concentrations of 25(OH)D may have an adverse effect on common complications of pregnancy, such as gestational hypertension (GH), preeclampsia (PE), and gestational diabetes mellitus (GDM). The aim of this study was to analyze the levels of sFlt-1 in Polish women with physiological pregnancies and pregnancies complicated by GH, PE, and GDM. Moreover, we analyzed relationships between the maternal serum sFlt-1 level and the sFlt-1 to 25(OH)D ratio and the risk of GH and PE. MATERIAL AND METHODS: The study included 171 women with complicated pregnancies; among them are 45 with GH, 23 with PE, and 103 with GDM. The control group was comprised of 36 women with physiological pregnancies. Concentrations of sFl-1 and 25(OH)D were measured before delivery, with commercially available immunoassays. RESULTS: Women with GH differed significantly from the controls in terms of their serum sFlt-1 levels (5797 pg/ml vs. 3531 pg/ml, p = 0.0014). Moreover, a significant difference in sFlt-1 concentrations was found between women with PE and those with physiological pregnancies (6074 pg/ml vs. 3531 pg/ml, p < 0.0001). GDM did not exert a statistically significant effect on serum sFlt-1 levels. Both logistic regression and ROC analysis demonstrated that elevated concentration of sFlt-1 was associated with greater risk of GH (AUC = 0.70, p = 0.0001) and PE (AUC = 0.82, p < 0.0001). Also, the sFlt-1 to 25(OH)D ratio, with the cutoff values of 652 (AUC = 0.74, p < 0.0001) and 653 (AUC = 0.88, p < 0.0001), respectively, was identified as a significant predictor of GH and PE. CONCLUSIONS: Determination of the sFlt-1/25(OH)D ratio might provide additional important information and, thus, be helpful in the identification of patients with PE and GH, facilitating their qualification for intensive treatment and improving the neonatal outcomes.

Expression of zinc finger transcription factors (ZNF143 and ZNF281) in serous borderline ovarian tumors and low-grade ovarian cancers.

Low-grade ovarian cancers represent up to 8% of all epithelial ovarian carcinomas (EOCs). Recent studies demonstrated that epithelial-mesenchymal transition (EMT) is crucial for the progression of EOCs. EMT plays a key role in cancer invasion, metastasis formation and chemotherapy resistance. An array of novel EMT transcription factors from the zinc finger protein family have been described recently, among them zinc finger protein 143 (ZNF143) and zinc finger protein 281 (ZNF281). The study included tissue specimens from 42 patients. Based on histopathological examination of surgical specimens, eight lesions were classified as serous borderline ovarian tumors (sBOTs) and 34 as low-grade EOCs. The proportions of the ovarian tumors that tested positively for ZNF143 and ZNF281 were 90 and 57%, respectively. No statistically significant differences were found in the expressions of ZNF143 and ZNF281 transcription factors in SBOTs and low-grade EOCs. Considering the expression patterns for ZNF143 and ZNF281 identified in this study, both sBOTs and low-grade EOCs might undergo a dynamic epithelial-mesenchymal interconversion. The lack of statistically significant differences in the expressions of the zinc finger proteins in sBOTs and low-grade serous EOCs might constitute an evidence for common origin of these two tumor types.

The analysis of human leukocyte antigen-G level in patients with endometrial cancer by Western blot technique.

PROBLEM: Aberrant expression of human leukocyte antigen-G (HLA-G) in various malignancies has been shown to participate in tumour development by suppressing immune regulation within the tumour microenvironment. The detection of HLA-G has reportedly been correlated with certain clinicopathological parameters in several neoplasms. Both the soluble and membranous forms of HLA-G are biologically active, and therefore, we aimed to evaluate the HLA-G level by Western blot technique. METHOD OF STUDY: The total amount of HLA-G protein was analyzed in the primary tumour in 113 tissue samples derived from patients with endometrial cancer. The HLA-G protein level was measured by Western Blot technique and was analyzed with respect to the clinicopathological parameters. RESULTS: Human leukocyte antigen-G protein levels were statistically significantly higher in the cancerous tissues derived from the women with advanced endometrial cancer than those from women with early stage disease. Moreover, we showed that endometrial cancer patients with lymph node metastases had statistically significantly higher HLA-G levels in the primary uterine tumour. CONCLUSION: The aberrant expression of HLA-G antigens by malignant cells could be one of the strategies tumour cells use to escape immune surveillance. The presence of HLA-G within the cancer nest and its microenvironment would seem to be linked to disease progression.

Expression of selected epithelial-mesenchymal transition transcription factors in serous borderline ovarian tumors and type I ovarian cancers.

Epithelial ovarian neoplasms are a heterogeneous group including tumor subsets with distinct clinicopathologic and molecular features. Recent evidence from molecular and genomic studies suggests that whereas low-grade serous carcinomas and high-grade serous carcinomas likely develop on two separate pathways, the low-grade serous carcinomas and serous borderline ovarian tumors may represent various stages of the same developmental continuum. The transformation of borderline ovarian tumors into an invasive neoplasm is associated with an array of molecular changes, inter alia controlled by p53 and PI3K/Akt pathway, as well as with a decrease in E-cadherin expression. The latter implies that epithelial-mesenchymal transition is a critical determinant of borderline ovarian tumor invasiveness. The aim of this study was to analyze the expression of transcription factors involved in epithelial-mesenchymal transition: SNAIL, SLUG, TWIST 1, TWIST 2, ZEB 1, and ZEB 2 in borderline tumors and type I ovarian cancers. The study included tissue specimens from 42 patients with histopathologically verified ovarian masses. The expressions for SLUG, TWIST 1, ZEB1, and ZEB 2 were scored based on the nuclear staining, and the expressions of SNAIL and TWIST 2 based on the cytoplasmic and/or nuclear staining. The proportions of ovarian tumors with the immunoexpression of the epithelial-mesenchymal transition transcription factors were 85.7% for SNAIL, 100% for SLUG, 9.5% for TWIST 1, 95.2% for TWIST 2, 23.8% for ZEB 1, and 0% for ZEB 2. The expression patterns of SNAIL, SLUG, TWIST, and ZEB identified in this study suggest that both serous borderline ovarian tumors and type I ovarian cancers undergo dynamic epithelial-mesenchymal interconversions. Our findings obtained in the two groups of tumors which shared some etiopathogenic pathways imply that the expression of the epithelial-mesenchymal transition transcription factors may be activated at early stages of the epithelial-mesenchymal transition, and thus these molecules may play a pivotal role in the development of both serous borderline ovarian tumors and type I ovarian cancer.

Testing for NRAS Mutations in Serous Borderline Ovarian Tumors and Low-Grade Serous Ovarian Carcinomas.

The Idylla NRAS Mutation Test, performed on the Biocartis Idylla system, is an in vitro diagnostic tool for the qualitative assessment of 18 NRAS mutations in codons 12, 13, 59, 61, 117, and 146. Low-grade serous ovarian cancer (LGSC) represents less than 10% of all serous ovarian carcinomas. LGSCs are believed to arise from preexisting cystadenomas or serous borderline tumors (SBOTs) that eventually progress to an invasive carcinoma. The molecular analysis of cancer-causing mutations and the development of targeted biological therapies constitute a milestone in the diagnosis and therapy of ovarian malignancies. According to some authors, NRAS may be an important oncogene for the progression of SBOT to a frankly invasive disease. The primary aim of this study was to verify if a fully integrated, real-time PCR-based Idylla system can be used for the rapid determination of the NRAS mutation status in patients with serous borderline ovarian tumors and low-grade serous ovarian carcinomas. The study included tissue specimens from 12 patients with histopathologically verified ovarian masses, operated on at the Department of Obstetrics and Gynecology, Nicolaus Copernicus University, Collegium Medicum in Bydgoszcz (Poland), between January 2009 and June 2012. The mean age of the study patients was 52.5 years (range 27-80 years). NRAS mutation in codon 13 (G13D, p.Gly13Asp; nucleotide: c.38G>A) was found in one patient, a woman with low-grade serous ovarian carcinoma. To the best of our knowledge, our experiment was the first published study using the novel Idylla NRAS Mutation Test for the evaluation of ovarian tumors in a clinical setting. The Idylla platform is an interesting ancillary first-line rapid and fully automated instrument to detect NRAS mutations in SBOTs and LGSCs. However, the clinical usefulness of this method still needs to be verified in larger groups of cancer patients.

KRAS mutation testing in borderline ovarian tumors and low-grade ovarian carcinomas with a rapid, fully integrated molecular diagnostic system.

Epithelial ovarian neoplasms are a heterogeneous group of tumors, including various malignancies with distinct clinicopathologic and molecular features. Mutations in BRAF and KRAS genes are the most frequent genetic aberrations found in low-grade serous ovarian carcinomas and serous and mucinous borderline tumors. Implementation of targeted therapeutic strategies requires access to highly specific and highly sensitive diagnostic tests for rapid determination of mutation status. One candidate for such test is fully integrated, real-time polymerase chain reaction-based Idylla™ system for quick and simple detection of KRAS mutations in formaldehyde fixed-paraffin embedded tumor samples. The primary aim of this study was to verify whether fully integrated real-time polymerase chain reaction-based Idylla system may be useful in determination of KRAS mutation status in patients with borderline ovarian tumors and low-grade ovarian carcinomas. The study included tissue specimens from 37 patients with histopathologically verified ovarian masses, operated on at the Department of Obstetrics and Gynecology, Nicolaus Copernicus University Collegium Medicum in Bydgoszcz (Poland) between January 2009 and June 2012. Based on histopathological examination of surgical specimens, 30 lesions were classified as low-grade ovarian carcinomas and 7 as borderline ovarian tumors. Seven patients examined with Idylla KRAS Mutation Test tested positive for KRAS mutation. No statistically significant association was found between the incidence of KRAS mutations and histopathological type of ovarian tumors. Mean survival of the study subjects was 48.51 months (range 3-60 months). Presence of KRAS mutation did not exert a significant effect on the duration of survival in our series. Our findings suggest that Idylla KRAS Mutation Test may be a useful tool for rapid detection of KRAS mutations in ovarian tumor tissue.

Determination of BRAF V600E (VE1) protein expression and BRAF gene mutation status in codon 600 in borderline and low-grade ovarian cancers.

Epithelial ovarian tumors are a group of morphologically and genetically heterogeneous neoplasms. Based on differences in clinical phenotype and genetic background, ovarian neoplasms are classified as low-grade and high-grade tumor. Borderline ovarian tumors represent approximately 10%-20% of all epithelial ovarian masses. Various histological subtypes of ovarian malignancies differ in terms of their risk factor profiles, precursor lesions, clinical course, patterns of spread, molecular genetics, response to conventional chemotherapy, and prognosis. The most frequent genetic aberrations found in low-grade serous ovarian carcinomas and serous borderline tumors, as well as in mucinous cancers, are mutations in BRAF and KRAS genes. The most commonly observed BRAF mutation is substitution of glutamic acid for valine in codon 600 (V600E) in exon 15. The primary aim of this study was to determine whether fully integrated, real-time polymerase chain reaction-based Idylla™ system may be useful in determination of BRAF gene mutation status in codon 600 in patients with borderline ovarian tumors and low-grade ovarian carcinomas. The study included tissue specimens from 42 patients with histopathologically verified ovarian masses, who were operated on at the Department of Obstetrics and Gynecology, Nicolaus Copernicus University Collegium Medicum in Bydgoszcz (Poland). Based on histopathological examination of surgical specimens, 35 lesions were classified as low-grade ovarian carcinomas, and 7 as borderline ovarian tumors. Specimens with expression of BRAF V600E (VE1) protein were tested for mutations in codon 600 of the BRAF gene, using an automated molecular diagnostics platform Idylla™. Cytoplasmic immunoexpression of BRAF V600E (VE1) protein was found in three specimens: serous superficial papilloma, serous papillary cystadenoma of borderline malignancy, and partially proliferative serous cystadenoma. All specimens with the expression of BRAF V600E (VE1) protein were tested positively for BRAF V600E/E2/D mutation. No statistically significant relationship (p > 0.05) was found between the presence of BRAF V600E mutation and the probability of 5-year survival. BRAF mutation testing with a rapid, fully integrated molecular diagnostics system Idylla™ may be also a powerful prognostic tool in subjects with newly diagnosed serous borderline tumors, identifying a subset of patients who are unlikely to progress.

High Quality Independent From a Donor: Human Amniotic Fluid Derived Stem Cells-A Practical Analysis Based on 165 Clinical Cases.

The aim of the study was to extend the potential use of human stem cells isolated from amniotic fluid in medical applications by confirming their high homogeneity and quality. Amniotic fluid samples were collected during amniocentesis from 165 women during pregnancy. The proliferation rate, clonogenicity, karyotype, aging process, pluripotent cell markers, expression of surface markers, and the potential to differentiate into adipose, bone and cartilage cells of hAFSCs were analyzed. Obtained results revealed that mesenchymal stem cells could be derived successfully from amniotic fluid, which exhibit properties comparable with MSCs of other origins. It is the first study, in which such a large group of patients was involved. Comprehensive statistical and biological analysis were conducted some of which clearly being innovative in relation to human amniotic fluid-derived stem cells. J. Cell. Biochem. 118: 116-126, 2017. © 2016 Wiley Periodicals, Inc.

Molecular diagnosis in type I epithelial ovarian cancer.

The term epithelial ovarian cancer (EOC) refers to a heterogeneous group of tumors, including serous, mucinous, endometrioid and clear cell carcinomas, each characterized by specific molecular background and clinical outcome. A growing body of evidence suggests that molecular pathogenesis of ovarian cancer involves two general pathways. The first pathway results in transformation of normal ovarian tissue to borderline tumors, which may further progress to low-grade serous, mucinous, endometrioid and clear cell carcinomas. Tumors from this group are characterized by slow proliferation, and approximately 55% 5-year survival rate. Type I tumors often harbor somatic mutations in protein kinase genes, as well as in genes for other signaling molecules. Both BRAF and KRAS mutations lead to a constitutive activation of their downstream target, mitogen-activated protein kinase. Identification of molecular profile may be crucial for the diagnosis of ovarian tumors, choice of adjuvant targeted therapy after primary cytoreductive treatment, or management of recurrence in patients with advanced type I epithelial ovarian neoplasms. Point mutations in cancer cells can be detected with many various methods. KRAS, NRAS and BRAF mutational status can be determined by Sanger sequencing (still considered a gold standard), as well as using numerous various techniques, such as allele-specific PCR, single nucleotide primer extension assays, pyrosequencing, real-time PCR, high-resolution melting curve analysis, amplification refractory mutation system, strip or chip assay combining PCR followed by hybridization to a KRAS or NRAS-specific probe, next-generation sequencing (NGS), and matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Application of direct sequencing as a routine method for cytological diagnosis used in a hospital setting requires expensive equipment and implementation of complicated procedures. Another factor limiting application of this method in everyday clinical practice are long analytical times. This stimulated search for a simple, rapid, specific and sensitive methodology to detect point mutations. Recently, some new molecular assays for the detection of BRAF, KRAS and NRAS mutations have become available. These are fully-automated molecular diagnostic systems for quantitative allele-specific RT-PCR-based analyses. Using this instrument, even pathologists from less experienced laboratories can easily integrate morphological findings with molecular data being crucial for further diagnostic and therapeutic decisions.

[Evaluation of carbetocin and oxytocin efficacy in prevention of postpartum hemorrhage in women after cesarean section]. / Ocena skutecznosci karbetocyny i oksytocyny w profilaktyce krwawien poporodowych u kobiet po cieciu cesarskim.

INTRODUCTION: Postpartum hemorrhage (PPH) constitutes the main cause of delivery-related maternal mortality worldwide. Identification of the risk factors, as well as knowledge about preventive measures and adequate management, allow to limit blood loss. Oxytocin, carbetocin, methylergometrine, dinoprostone, suiprostone, and misoprostol are commonly used drugs in prevention of PPH. OBJECTIVES: The aim of the study was to evaluate the efficacy of carbetocin and oxytocin in prevention of PPH after cesarean section. MATERIAL AND METHODS: The study included 130 female patients after C-section who received 1 00 pg of carbetocin i.v. as a preventive agent after the surgery The control group consisted of 60 women who received 10 units of oxytocin i.v. In the study the risk factors for PPH were determined, and hemoglobin and hematocrit values before and 12 hours after birth, as well as blood loss and the need to use other prevyentfive and operational methods were evaluated. Results were compared between the groups. Statistical analysis was performed with the use of Statistica for hemoglobin and hematocrit values. The p-value of < or = 0.05 was considered as statistically significant. RESULTS: Risk factors for PPH occurred in almost 100% of the women with carbetocin and in 90% of the women with oxytocin. The decrease in hemoglobin and hematocrit levels was not statistically significant, although a greater drop was detected in the group with oxytocin (hemoglobin - 1.24 vs. 1.17 g%, hematocrit - 3.26 vs. 2.93%). The decrease in hematological values was not statistically significant between both groups. In the group with'carbetocin, there was no need for additional pharmacological therapy or operative procedures. No adverse events in either of the groups were observed. CONCLUSIONS: (1.) Carbetocin effectively prevents PPH after C-section. (2.) Carbetocin seems to have high efficiency in PPH prevention in pregnant women classified to the PPH risk group. (3.) Efficacy of Carbetocin in PPH prevention is higher than oxytocin.

[Indications for genetic amniocentesis investigated at the Department of Gynecology, Obstetrics, and Oncologic Gynecology, Nicolaus Copernicus University, Collegium Medicum, Bydgoszcz]. / Analiza wskazan do amniopunkcji genetycznej w zaleznosci od wieku pacjentek na podstawie materialu Kliniki Poloznictwa, chorób Kobiecych i Ginekologii Onkologicznej CM UMK w Bydgoszczy.

INTRODUCTION: Genetic amniocentesis (GA) is the most common prenatal diagnostic test. One of the main indications for GA is maternal age of > or = 35 years. In many countries, the age indication has been replaced by an assessment of individual risk for chromosomal abnormalities, calculated on the basis of maternal age, pregnancy duration, as well as a combination of biochemical and ultrasound markers. OBJECTIVES: The aim of the study was to investigate indications for and results of GA performed between 2010-2012 at the Department of Gynecology Obstetrics, and Oncologic Gynecology Nicolaus Copernicus University Collegium Medicum, Bydgoszcz. MATERIALS AND METHODS: A total of 632 GA tests were performed at the Department of Gynecology Obstetrics, and Oncologic Gynecology Nicolaus Copernicus University Collegium Medicum, Bydgoszcz. Average maternal age was 34 (between 17 and 47 years), with patients < 35 constituting 47.9% (N = 303), and patients > or = 35 constituting 52.1% (N = 329) of the investigated group. Indications for GA as well as test results were analyzed in relation to maternal age. The result of earlier non-invasive tests were also analyzed. RESULTS: Abnormal ultrasound findings, combined with abnormal first-trimester screening results, were the most common indication (46.53%) for GA in patients < 35 years, whereas abnormal first-trimester screening results, combined with a history of obstetric complications, were the reason for GA in patients > or = 35 years. Mean time of GA was 16 gestational weeks in both groups. Abnormal karyotype was detected in 74 (11.7%) cases. 13 or any other abnormal karyotypes occurrence were observed in both age groups. GA-related complications (miscarriage/intrauterine fetal death) occurred in 9 (1.42%) cases. CONCLUSIONS: If performed properly GA between 15 and 20 weeks of pregnancy is a harmless procedure both, for the mother and the fetus, associated with an acceptable complication rate. Prenatal screening for the most common malformations and chromosomal aberrations should be offered to all pregnant women in Poland, regardless of their age.

[Clinical study of perinatal hysterectomy between 2000-2011 in the clinic of obstetrics, gynecological diseases and oncological gynecology in Bydgoszcz]. / Analiza kliniczna okoloporodowego wyciecia macicy w latach 2000-2011 w Klinice Poloznictwa, Chorób Kobiecych i Ginekologii Onkologicznej w Bydgoszczy.

INTRODUCTION: Perinatal hysterectomy (PH) is usually a life-saving procedure, which is performed after all conservative treatment options fail. The PH frequency rate ranges from 0.04 to 0.23%. The most frequent indications for this procedure include: abnormal placental implantation, placenta previa, uterine rupture and uterine atony OBJECTIVE: Clinical study of perinatal hysterectomy cases taking into consideration the frequency indications, complications and risk factors related to this procedure. MATERIALS AND METHODS: The study included 16 women who underwent perinatal hysterectomy at the Department and Clinic of Obstetrics and Gynecological Diseases between 2000-2011. The following data were collected from medical records: course of pregnancy labor and puerperium. The profile of the study group was conducted in terms of: maternal age, parity gestation length, history of caesarean sections and gynecological operations. The following factors were studied: the termination of pregnancy, indications for caesarean section, hysterectomy-related complications and indications, neonatal birth weight and Apgar score. The statistical analysis was performed using Statistica 9.1 by StatSoft. Data are expressed as the arithmetic mean and standard deviation (SD). RESULTS: Sixteen perinatal hysterectomy procedures were performed, accounting for 0.066% of the overall number of labors. Average maternal age and pregnancy length were 31.6 years [SD+/-6.3] and 36.1 weeks of gestation [SD+/-3.4], respectively PH was more frequently performed among multiparous women (81.25%) and after caesarean sections (87.5%). Fetal asphyxia was the most frequent indication for caesarean section (35.7%). Fourteen percent of all indications accounted for the lack of consent from a pregnant woman to make an attempt at spontaneous vaginal delivery after previous c-section. Fifty percent of the women from the study group had a previous caesarean section, whereas 25% had more than one prior c-section. Between 2009-2011, as compared to previous years, the highest percentage of hysterectomies (80%) was reported in pregnant women after a previous caesarean section. The most frequent indication for hysterectomy included abnormal placental implantation (43.75%) diagnosed more often in patients with a history of caesarean section (57%). Among PH complications, a hemorrhagic shock was reported in 37.4% and bladder injury in 18.7% of the women. Every patient required a transfusion of erythrocyte concentrate, 4.7 units [SD+/-3.5] on average. Twenty-five percent of the neonates were born in poor condition with an Apgar score of 1-3. In case of all women, the therapy required cooperation of different specialists including obstetricians, anesthesiologists, urologists, surgeons and general practitioners. CONCLUSIONS: 1. Current and previous caesarean section constitutes a risk factor for perinatal hysterectomy 2. Placental pathology is the most frequent indication for perinatal hysterectomy 3. The growing number of caesarean sections should encourage obstetricians to conduct a more careful analysis of indications.

The role of Hypoxia-inducible factor-1 α , glucose transporter-1, (GLUT-1) and carbon anhydrase IX in endometrial cancer patients.

Hypoxia-inducible factor-1α (HIF-1α), glucose transporter-1 (GLUT-1), and carbon anhydrase IX (CAIX) are important molecules that allow adaptation to hypoxic environments. The aim of our study was to investigate the correlation between HIF-1α, GLUT-1, and CAIX protein level with the clinicopathological features of endometrial cancer patients. Materials and Methods. 92 endometrial cancer patients, aged 37-84, were enrolled to our study. In all patients clinical stage, histologic grade, myometrial invasion, lymph node, and distant metastases were determined. Moreover, the survival time was assessed. Immunohistochemical analyses were performed on archive formalin fixed paraffin embedded tissue sections. Results. High significant differences (P = 0.0115) were reported between HIF-1α expression and the histologic subtype of cancer. Higher HIF-1α expression was associated with the higher risk of recurrence (P = 0.0434). The results of GLUT-1 and CAIX expression did not reveal any significant differences between the proteins expression in the primary tumor and the clinicopathological features. Conclusion. The important role of HIF-1α in the group of patients with the high risk of recurrence and the negative histologic subtype of the tumor suggest that the expression of this factor might be useful in the panel of accessory pathomorphological tests and could be helpful in establishing more accurate prognosis in endometrial cancer patients.

The preoperative maximum standardized uptake value measured by 18F-FDG PET/CT as an independent prognostic factor of overall survival in endometrial cancer patients.

PURPOSE: The aim of this study was to determine if the preoperative maximum standardized uptake value (SUVmax) measured by 18F-FDG PET/CT in the primary tumor has prognostic value in the group of patients with endometrial cancer. PATIENTS, MATERIALS, AND METHODS: A total of one hundred one consecutive endometrial cancer patients, age range 40-82 years (mean 62 years) and FIGO I-IV stage, who underwent 18-FDG-PET/CT within two weeks prior radical surgery, were enrolled to the study. The maximum SUV was measured and compared with the clinicopathologic features of surgical specimens. The relationship between SUVmax and overall survival was analyzed. RESULTS: The mean preoperative SUVmax was 14.34; range (3.90-33.80) and was significantly lower for FIGO I than for higher stages (P = 0.0012), as well as for grade 1 than for grade 2 and 3 (P = 0.018), deep myometrial invasion (P = 0.0016) and for high risk group (P = 0.0004). The analysis of survival ROC curve revealed SUVmax cut-off value of 17.7 to predict high risk of recurrence. Endometrial cancer patients with SUVmax higher than 17.7 characterized by lower overall survival. CONCLUSION: The preoperative SUVmax measured by 18F-FDG PET/CT is considered as an important indicator reflecting tumor aggressiveness which may predict poor prognosis. High value of SUVmax would be useful for making noninvasive diagnoses and deciding the appropriate therapeutic strategy for patients with endometrial cancer.

Human amniotic-fluid-derived stem cells: a unique source for regenerative medicine.

INTRODUCTION: The first application of tissue engineering was based on the use of differentiated cells from the adult organism, which was associated with an invasiveness and high risk of diseased cells' transplantation. Over the years, the range of available cell populations for tissue engineering has widened. AREAS COVERED: We review the comprehensive information concerning the characteristic features of amniotic-fluid-derived stem cells (AFSCs). We also review the potential applications of these cells in clinical practice. EXPERT OPINION: AFSCs hold promise for the future treatment of many incurable diseases. However, such cell-based therapies have some limitations, and there are questions relating to the use of stem cells, which should be carefully analyzed before translation of these cells into clinical practice.

Serum inhibin A and inhibin B levels in epithelial ovarian cancer patients.

THE AIM: of our study was to examine serum inhibin A and inhibin B concentrations in ovarian cancer patients in relation to clinicopathological features and 5-year survival. MATERIAL AND METHODS: We enrolled 90 epithelial ovarian cancer patients in our study, aged 45-81 years, who underwent optimal cytoreductive surgery. In all patients, serum inhibin A and inhibin B concentrations were measured using a two-step sandwich type enzyme immunoassay before surgery. RESULTS: In the group of patients with ovarian cancer median serum concentration of inhibin A was 3.87 pg/mL (0.96-10.09) and inhibin B was 13.9 pg/mL (5.1-45.0). Median concentrations of inhibin A and B in relation to FIGO stage and histological subtype did not differ significantly. Inhibin A levels were significantly higher in patients with lower grading (G1 and G2) in comparison to those with higher grade G3 (p=0.001). There were no differences in inhibin B concentrations in relation to grading. The Kaplan-Meier analyses demonstrated no differences in survival rate in relation to inhibin A levels, while there was a stepwise impairment of 5-years survival with increased inhibin B level. In the group of patients with inhibin B levels higher than 20 pg/ml the survival rate was lower (p=0,00625, log-rank test). CONCLUSION: 1. Higher inhibin A serum levels were found in patients with highly differentiated ovarian carcinoma compared to the group of patients with a poorly differentiated cancer, which may confirm the influence of inhibin A on cell proliferation processes. 2. A significant importance of inhibin B was demonstrated in the prediction of death within less than a five year period. The probability of survival in patients featuring high inhibin B levels was lower with statistical significance. This may indicate the need for further studies on how to block the inhibin B activation pathway in the ovarian carcinoma therapy.

Stromal derived factor-1 (SDF-1) and its receptors CXCR4 and CXCR7 in endometrial cancer patients.

PURPOSE: One of the most important function of stromal derived factor-1 (SDF-1) and its receptors, is regulating the process of metastasis formation. The aim of our study was to investigate the correlation between SDF-1, CXCR4 and CXCR7 protein levels measured by immunohistochemistry with the clinicopathological features and the survival of endometrial cancer patients. MATERIALS AND METHODS: 92 patients aged 37-84 (mean 65.1±9.5) were enrolled to our study between January 2000 and December 2007. After the diagnosis of endometrial cancer, all women underwent total abdominal hysterectomy, with bilateral salpingoophorectomy and pelvic lymph node dissection. In all patients clinical stage (according to FIGO classification), histologic grade, myometrial invasion, lymph node and distant metastases were determined.Furthermore, the survival time was assessed. Immunohistochemical analyses of SDF-1, CXCR4 and CXCR7 were performed on archive formalin fixed paraffin embedded tissue sections. RESULTS: Statistically significant correlations (p<0.01) were reported between SDF-1 and the clinical stage of disease, lymph node metastases, distant metastases, deep myometrial invasion (≥50%), cervical involvement, involvement of adnexa. Statistically significant correlation (p<0.01) was found between SDF-1 expression and the risk of the recurrence. Higher SDF-1 expression was associated with a higher risk of recurrence (p = 0.0001). The results of CXCR4 and CXCR7 expression didn't reveal any significant differences(p>0.05) between the proteins expression in the primary tumor cells and the clinicopathological features. Moreover, the Kaplan-Meier analyses demonstrated a stepwise impairment of cancer overall survival (OS) with increasing SDF-1 expression. CONCLUSION: The important role of SDF-1 as a predictor of negative clinicopathological characteristics of a tumor suggests that the expression of this stromal factor should be included in the panel of accessory pathomorphological tests and could be helpful in establishing a more accurate prognosis in endometrial cancer patients.

[SUVmax measured by 18F FDG PET/CT in the primary tumor in relation to clinical and pathological features of endometrial cancer]. / Wartosci SUVmax mierzone za pomoca 18F FDG PET/CT w guzie pierwotnym a cechy kliniczno-patologiczne endometrioidalnego raka endometrium.

THE AIM: The aim of the study was to determine the value of SUVmax by PET/CT measured before surgery with special focus on FIGO stage, myometrial invasion, grading and risk stratification. METHODS: A total of 90 women, aged 37-84 (mean 65.1 +/- 9.5) with endometrial cancer (FIGO I and II) underwent 18F FDG PET/CT imaging before surgery SUVmax of the primary tumor was assessed and compared with histological prognostic factors (FIGO stage, grading, myometrial invasion, risk group). RESULTS: The mean SUVmax in the study group was 13.95 +/- 5.49. SUVmax was significantly higher with high FIGO stage (p=0,0009), deep myometrial invasion (p=0.0005), high grade (p=0.0331) and high risk tumors (0.0007). Patients with the poor prognosis had significantly higher SUVmax values. CONCLUSIONS: The preoperative SUVmax measurements of the primary tumor of endometrial cancer may give additional clinical and prognostic information about risk factors and poor prognosis. High value of SUVmax might be useful in making noninvasive diagnoses and deciding the appropriate therapeutic strategy for patients with endometrial cancer However there is not enough evidence that it could in fact replace surgical staging.

Serum anti-müllerian hormone levels in patients with epithelial ovarian cancer.

Objectives. The aim of our study was to examine serum anti-Müllerian hormone (AMH) concentration in ovarian cancer patients in relation to clinicopathological features, such as a pathological subtype of the tumor, (FIGO) stage, grading, and overall 5-year survival. Material and Methods. We enrolled 72 epithelial ovarian cancer patients in our study, aged 45-79 years, who underwent optimal cytoreductive surgery. In all patients, serum AMH concentration was measured using a two-step sandwich type enzyme immunoassay before surgery. As a reference value for women over 45 years we accepted anti-Müllerian hormone concentration below 1 ng/mL. Results. In the whole group of patients with ovarian cancer, median serum concentration of AMH was 0.07 (0.0-0.37) ng/mL, whereas in the group of those with positive AMH values (≥0.14 ng/mL) it was 0.31 (0.15-0.73) ng/mL. No significant correlation was found between serum AMH levels and FIGO stage, histological subtype, or grading (P > 0.05). The analysis of five-year survival rate related to AMH levels showed no statistically significant differences. There were no differences in survival rates between patients with positive or negative serum AMH levels. Conclusion. Measurement of serum anti-Müllerian hormone levels was not useful in predicting clinicopathological features and survival in patients with ovarian cancer.