Maternal Plasma Metabolomic Profiles in Spontaneous Preterm Birth: Preliminary Results.

OBJECTIVE: To profile maternal plasma metabolome in spontaneous preterm birth. METHOD: In this retrospective case-control study, we have examined plasma of patient with preterm birth (between 22 and 36 weeks of pregnancy (n = 57)), with threatened preterm labor (between 23 and 36 weeks of pregnancy (n = 49)), and with term delivery (n = 25). Plasma samples were analysed using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) in positive and negative polarity modes. RESULTS: We found 168 differentially expressed metabolites that were significantly distinct between study groups. We determined 51 metabolites using publicly available databases that could be subdivided into one of the five groups: amino acids, fatty acids, lipids, hormones, and bile acids. PLS-DA models, verified by SVM classification accuracy, differentiated preterm birth and term delivery groups. CONCLUSIONS: Maternal plasma metabolites are different between term and preterm parturitions. Part of them may be related with preterm labor, while others may be affected by gestational age or the beginning of labor. Metabolite profile can classify preterm or term delivery groups raising the potential of metabolome as a biomarker to identify high-risk pregnancies. Metabolomic studies are also a tool to detect individual compounds that may be further tested in targeted researches.

Angiogenic factor screening in women with mild preeclampsia - New and significant proteins in plasma.

INTRODUCTION: The aim of this study was to analyse a panel of 60 angiogenic factors (pro-angiogenic and antiangiogenic) in the plasma of women with mild preeclampsia. MATERIALS AND METHODS: We recruited 21 women between 25 and 40 weeks gestation with diagnosed mild preeclampsia into the study group and 27 healthy women with uncomplicated pregnancies of corresponding gestational age to that of the study to the control group. We used a quantitative protein macroarray method that allowed for analysis of 60 angiogenic proteins per sample simultaneously. RESULTS: We showed a statistically significant increase in the concentration of 8 proteins, interferon gamma (IFN-γ), interleukin 6 (IL-6), leukaemia inhibitory factor (LIF), heparin-binding EGF-like growth factor (HB-EGF), hepatocyte growth factor (HGF), C-X-C motif chemokine 10 (IP-10), leptin and platelet-derived growth factor BB (PDGF-BB), as well as a significant decrease in the concentration of 3 proteins, vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and follistatin, in the plasma of women with preeclampsia. CONCLUSION: Based on our findings, it seems that protein factors may play an important role in the pathogenesis of preeclampsia, and there are many proteins that have not been studied in PE to date. There are no previous studies assessing the LIF, follistatin, HGF, HB-EGF and PDGF-BB concentrations in the plasma of women with PE; therefore, our obtained results indicate that these proteins are new factors that can play an important role in the pathomechanisms of PE.

Identification of preterm birth based on RQA analysis of electrohysterograms.

BACKGROUND AND OBJECTIVE: Common methods for data analysis are mainly based on linear concepts, but in recent years nonlinear dynamics methods have been introduced. It is a well-known fact that In typical biological systems lack of stationarity and rather sudden changes of state are the properties distinguishing them from each other. There is an urgent need to better understand the mechanical activity of the myometrium (its contractility) to find a solution for preterm delivery problem, the largest cause of neonatal deaths and morbidity. The electrohysterographic signal (EHG) is a good non-linear, bioelectrical indicator for the detection and identification of term and preterm birth. METHODS: The material of the study consists of EHG signals, obtained from 20 patients between the 24th and the 28th week of pregnancy with threatened preterm labor. The women were divided into two groups: those delivering after more than 7 days - group A (n = 10) and women delivering within 7 days - group B (n = 10). In this paper, an analysis of bioelectrical signals was performed by recurrence quantification analysis (RQA) and principal component analysis (PCA) to distinguish particular patterns for term and preterm birth. To date, these methods have not been used for the evaluation of bioelectrical activity in the uterus. To train novel classifiers for the EHG signals Support Vectors Machine classifications (multiclass SVM) was used. Statistical analysis was performed by means of non-parametric Mann-Whitney test. RESULTS: From among eleven parameters obtained from recurrence quantification analysis, five most appropriate were chosen: Recurrence Rate, Determinism, Laminarity, Entropy and Recurrence Period Density Entropy. Significant increase (p < .001) of Recurrence Rate was found in patients from group B, while increase of parameters, besides Laminarity, was found in patients from group A. The accuracy of classification obtained as a result of the analysis increased to 83,32%. CONCLUSION: We showed that the respectively selected recurrence quantificators obtained for that time series could be used to classify all those signals to the appropriate group. The proposed analysis could help in detecting preterm labor based on the EHG signal dynamics.

Profiling of selected angiogenesis-related genes in serous ovarian cancer patients.

PURPOSE: Since angiogenesis plays an important role in the pathogenesis of ovarian cancer the aim of the study was to compare the expression of the most relevant angiogenesis-related genes in serous ovarian cancer samples. Genes were divided into 5 subgroups according to their angiogenic potential: growth factors and their receptors; cytokines/chemokines; adhesion molecules and other matrix related proteins; transcriptions factors and signaling molecules; morphogenic factors, and angiogenesis inhibitors. MATERIALS/METHODS: Twenty-nine patients were involved in the study: 20 with serous ovarian cancer and 9 healthy controls. All neoplasms were confirmed by histopathological examination. Healthy ovarian control samples were obtained from women diagnosed with fibroids and had previously scheduled operations. Real-time PCR gene arrays were used to examine the expression of 84 human angiogenesis-related genes and expression of selected proteins was assessed with ELISA. RESULTS: Significantly higher expressions of 46 genes were found in the ovarian cancer group compared to the healthy control group. By the use of ELISA we confirmed the expression of three proteins i.e.: angiopoietin-2, angiopoietin-like protein 3, and angiopoietin receptor 2. Only angiopoietin-2 and angiopoietin receptor 2 showed significant differences between ovarian cancer and healthy controls. CONCLUSIONS: Changes in the expression of selected genes associated with angiogenesis may add new information to the pathogenesis of ovarian cancer. Although the angiopoietin-2 signaling pathway may play an important role in neovascularization in ovarian cancer, the role of angiopoietin-like protein 3 is yet to be established.

The role of vitamin D in reproductive dysfunction in women - a systematic review.

Vitamin D is essential for the proper functioning of the human body. There is also evidence of its strong association with fertility problems in women. This review aims to evaluate the relationship between vitamin D and diseases affecting women's fertility (polycystic ovarian syndrome (PCOS), uterine leiomyomas and endometriosis), and in vitro fertilization (IVF) outcome. A systematic review of the literature was conducted in Scopus and PubMed for relevant English language publications since 1989. Vitamin D influences the functioning of the reproductive system in women and has been associated with PCOS, uterine leiomyomas, endometriosis and in vitro fertilization (IVF) outcome. However, further studies on larger groups of patients are needed to establish what role vitamin D plays in the treatment of female infertility.

[First in the world application of next generation sequencing in preimplantation genetic diagnostics in clinical practice - a case report]. / Pierwsze w swiecie zastosowanie sekwencjonowania nastepnej generacji w diagnostyce przedimplantacyjnej w praktyce klinicznej - opis przypadku.

Preimplantation genetic diagnosis (PGD) is a well established method for detecting genetic abnormalities during the course of infertility treatment, resulting in thousands of healthy newborns delivered worldwide. PGD with next generation sequencing (NGS) provides new possibilities for diagnosis and new parameters for evaluation. The use of next-generation DNA sequencing technique has lead to great progress in the human genome analysis. The aim of this study was molecular analysis using next generation sequencing technique of embryos from a couple suffering from recurrent pregnancy losses. As a result of in vitro fertilization procedure, seven embryos were created. Seven blastomeres, one from each embryo, were analyzed. Transfer of two blastocysts in a fresh cycle resulted in the singleton pregnancy. Healthy baby girl was delivered via caesarean section after 28 weeks of gestation (weight: 1250g, Apgar score: 8/9). The reason for the premature labor was likely caused by mother's pneumonia. This is the first case of clinical use of the NGS in PGD in fresh cycle after blastomere biopsy.

Plasma C16-Cer levels are increased in patients with preterm labor.

INTRODUCTION: The pathogenesis of preterm labor is fragmentarily explained. The most widely accepted theory points out to infection and inflammation as possible causes, which can be mediated by potentially different factors, including sphingolipid mediators. Sphingolipids are a class of lipids that have been shown as important mediators in various cell processes such as: proliferation, growth, apoptosis, stress response, necrosis and inflammation. The aim of the study was to assess plasma concentrations of selected sphingolipids in patients with preterm labor. MATERIAL AND METHODS: We used ultra-high performance liquid chromatography with triple mass spectrometry (UHPLC-ESI-MS/MS) to assess plasma concentrations of the 11 sphingolipids in patients presenting with symptoms of preterm labor (n=61) and threatened preterm labor (n=40). RESULTS: We observed a statistically significant increase (p-value<0.004) in plasma concentrations of C16-Cer in patients with preterm labor as compared to the control group. We also found C16-Cer to be the best predictor of preterm labor in the group of patients with symptoms occurring after 32 weeks of gestation. CONCLUSIONS: Our findings show a possible involvement of selected sphingolipids, especially C16-Cer, in the pathogenesis of preterm labor. Their role as predictors of preterm delivery needs to be validated in the future on larger group of patients.

Differences in permeability of preterm and term fetal membranes to calcium ions - Preliminary report.

The metabolic activity of amniochorion contributes to the control of activation of labor-type uterine contractions. The study presents an experimental model of transport of calcium ions across the human amniochorion sampled directly after cesarean section in patients delivering both at term and prematurely. Transmembrane transport of calcium ions was lower in preterm vs. term tissue samples. The differences in permeability were most pronounced in the first 60 min of experiments. The results of the study provide evidence for the existence of an active mechanism of calcium transport which can contribute to regulating the contractility of the uterus.

Reproductive outcomes of intracytoplasmic sperm injection (ICSI) in good-prognosis patients who electively decided to limit the number of oocytes used for microinjection: a two-center comparative study.

OBJECTIVES: The aim of the study was to compare the outcomes of intracytoplasmic sperm injection/embryo transfer (ICSI/ET) between two IVF centers with similar pregnancy rates and embryo transfer policy but with two different approaches to good-prognosis patients who intentionally chose to limit the number of oocytes used for ICSI. MATERIAL AND METHODS: It was a retrospective two-center comparative study A total of 218 patients after successful retrieval of >10 mature oocytes following ovarian hyperstimulation were included in the study The number of fertilized oocytes used during ICSI/ET was limited to 6 and 10 in 108 and 110 patients of the Centre for Reproductive Medicine KRIOBANK and VitroLive Fertility Clinic, respectively RESULTS: No significant differences in the implantation rate (29.93% vs. 29.54%; p=0.94) and ongoing pregnancy rate (39.81% vs. 45.45%, p=0.40) were observed between patients who electively fertilized 6 as compared to 10 oocytes, respectively However in patients who deliberately limited the number of fertilized oocytes to 6 the following were observed: i) significantly fewer embryos available for ET (2.89 ± 1.23 vs. 3.77 ± 1.48, p<0.0 1); ii) considerably lower number of frozen embryos per cycle (1.05 ± 1.30 vs. 2.00 ± 1.67, p<0.01), and iii) lower rates of cycles with embryo cryopreservation (4 7.22% vs. 72.72%, p<0.01) as compared to patients with 10 fertilized oocytes. CONCLUSIONS: Elective fertilization of 6 vs. 10 oocytes does not adversely affect fresh ICSI/ET outcome in normal-responding patients. Restricted number of oocytes used for ICSI/ET may be a favorable alternative for couples who do not wish to cryopreserve surplus human embryos.

Increased Maternal and Cord Blood Betatrophin in Gestational Diabetes.

AIM: The aim of the study was to compare maternal and cord blood levels of betatrophin--a new peptide potentially controlling beta cell growth--as well as in its mRNA expression in subcutaneous adipose tissue, visceral adipose tissue and placental tissue obtained from pregnant women with normal glucose tolerance (NGT) and gestational diabetes (GDM). METHODS: Serum betatrophin and irisin concentrations were measured by ELISA in 93 patients with GDM and 97 women with NGT between 24 and 28 week of gestation. Additionally, maternal and cord blood betatrophin and irisin, as well as their genes (C19orf80 and Fndc5) expression were evaluated in 20 patients with GDM and 20 women with NGT at term. RESULTS: In both groups, serum betatrophin concentrations were significantly higher in the patients with GDM than in the controls (1.91 [1.40-2.60] ng/ml vs 1.63 [1.21-2.22] ng/ml, p=0.03 and 3.45 [2.77-6.53] ng/ml vs 2.78 [2.16-3.65] ng/ml, p=0.03, respectively). Cord blood betatrophin levels were also higher in the GDM than in the NGT group (20.43 [12.97-28.80] ng/ml vs 15.06 [10.11-21.36] ng/ml, p=0.03). In both groups betatrophin concentrations in arterial cord blood were significantly higher than in maternal serum (p=0.0001). Serum irisin levels were significantly lower in the patients with GDM (1679 [1308-2171] ng/ml) than in the healthy women between 24 and 28 week of pregnancy (1880 [1519-2312] ng/ml, p=0.03). Both C19orf80 and Fndc5 mRNA expression in fat and placental tissue did not differ significantly between the groups studied. CONCLUSIONS: Our results suggest that an increase in maternal and cord blood betatrophin might be a compensatory mechanism for enhanced insulin demand in GDM.

A predictive model for blastocyst formation based on morphokinetic parameters in time-lapse monitoring of embryo development.

PURPOSE: The aim of the study was to create a predictive model of blastocyst development based on morphokinetic parameters of time-lapse embryoscope monitoring. METHODS: Time-lapse recordings of 432 embryos (obtained from 77 patients), monitored in Embryoscope, were involved in the study. Patients underwent in vitro fertilization according to standard procedure between June 2012 and April 2013. A retrospective analysis of morphokinetic features, focused on duration of time from the Intracytoplasmic Sperm Injection (ICSI) procedure to consecutive embryo division for 2, 3, 4 and 5 blastomeres, as well as time intervals between each division, was conducted. All embryos were observed for 5 days. RESULTS: Based on the distribution of analyzed morphokinetic parameters and number of embryos developed to blastocyst, a range denoting the possibility of an embryo reaching blastocyst stage was determined. According to the obtained results, univariate and multivariate logistic regression analyses were performed. Based on the times of division for two and five blastomeres and intervals between the second and third division, a multivariate predictive model was created. The predictive equation was constructed based on the parameters of logistic regression analysis (odds ratios). Statistically significant differences (p < 0.001) in the size of the prediction parameter between the group of embryos developed to blastocyst (the median value: Me = 9.95, and quartiles: Q1 = 7.59, Q3 = 12.30) and embryos that did not develop to the blastocyst stage (Me = 4.66, Q1 = 2.33, Q3 = 8.19) were found. A Receiver Operating Characteristic (ROC) curve was created for the constructed predictive model. The Area Under the Curve was AUC = 0.806 with a 95 % confidence interval (0.747, 0.864). The predictive model constructed in this study has been validated using an independent data set, which indicates that the model is reliable and repeatable. CONCLUSIONS: Time-lapse imaging presents a new diagnostic tool for parametric evaluation of embryo development, from the oocyte stage, through fertilization, up to the blastocyst stage. The assessment of morphokinetic parameters can help us to provide more accurate information about the reproductive potential of embryos. It allows for early selection of embryos with high reproductive potential and shortens embryo incubation.

Comparison of the second-generation Beckman Coulter IVD and first-generation AnshLabs ELISA assays for anti-Müllerian hormone in patients undergoing IVF treatment.

OBJECTIVES: Ovarian reserve is the main factor influencing the efficacy of infertility treatment. Currently the anti- Müllerian hormone is the main indicator of the ovarian reserve and has a wide spectrum of clinical importance. It achieved a high clinical value right after the introduction of the first commercial AMH assays in 2005. Lack further research and development of the tests and monopoly on their production have led to a significant reduction of their quality resulting in lowered veracity and usefulness. Therefore, we searched for an alternative to the Beckman Coulter assay. The objective of the study was to draw a comparison between the commonly used second-generation assay by Beckman Coulter and the ultra-sensitive first-generation assay by AnshLabs. MATERIALS AND METHODS: Serum samples (n=520) were collected from female patients undergoing routine AMH evaluation before entering an IVF program. We chose samples of patients with the lowest correlation between the AMH serum level and response to stimulation. The AMH serum levels of the patients were examined using two AMH tests, the second-generation assay by Beckman Coulter and the first-generation assay by AnshLabs. Precision and accuracy of both methods were determined and the results of AMH serum levels of 130 patients were correlated with the number of: antral follicles (AFC), follicles after stimulation, and the obtained cumulus cells. RESULTS: Both precision and accuracy of the compared methods were highly satisfactory. The coefficients of variation obtained in the study conducted on two different levels of control material were lower than 12% and the load did not exceed 9%. The study proved that both of the methods yielded comparable results. The coefficient of variation between the first-generation and the second-generation AMH assays was 0.871. CONCLUSION: Both methods might be applied in the evaluation of the ovarian reserve. The first- and second-generation assays show comparable correlation with the clinical effects of stimulation, however it seems that first-generation assays are a better alternative to the unstable second-generation kits. The results from the first-generation assays are distributed on a wider range, which facilitates clinical interpretation.

Chemokines profiling of patients with preterm birth.

INTRODUCTION: Nowadays it is thought that the main cause of premature birth is subclinical infection. However, none of the currently used methods provide effective prevention to preterm labor. The aim of the study was to determine the concentration of selected chemokines in sera of patients with premature birth without clinical signs of infection (n = 62), threatened preterm labor (n = 47), and term births (n = 28). METHOD: To assess the concentration of chemokines in the blood serum, we used a multiplex method, which allows the simultaneous determination of 40 chemokines per sample. The sets consist of the following chemokines: 6Ckine/CCL21, Axl, BTC, CCL28, CTACK/CCL27, CXCL16, ENA-78/CXCL5, Eotaxin-3/CCL26, GCP-2/CXC, GRO (GRO α /CXCL1, GRO ß /CXCL2 and GRO γ /CXCL3), HCC-1/CCL14, HCC-4/CCL16, IL-9, IL-17F, IL18-BPa, IL-28A, IL-29, IL-31, IP-10/CXCL10, I-TAC/CXCL11, LIF, LIGHT/TNFSF14, Lymphotactin/XCL1, MCP-2/CCL8, MCP-3/CCL7, MCP-4/CCL13, MDC/CCL22, MIF, MIP-3 α /CCL20, MIP-3- ß /CCL19, MPIF-1/CCL23, NAP-2/CXCL7, MSP α , OPN, PARC/CCL18, PF4, SDF-1/CXCL12, TARC/CCL17, TECK/CCL25, and TSLP. RESULTS: We showed possible implication of 4 chemokines, that is, HCC-4, I-TAC, MIP-3 α , and TARC in women with symptoms of preterm delivery. CONCLUSION: On the basis of our findings, it seems that the chemokines may play role in the pathogenesis of preterm labor. Defining their potential as biochemical markers of preterm birth requires further investigation on larger group of patients.

Assessment of the selected biochemical markers in predicting preterm labour.

OBJECTIVE: The differential diagnosis between preterm and false labour remains one of the most challenging issues in perinatal medicine. AIM: To assess the prognostic importance of the selected biochemical markers in predicting preterm labour. MATERIAL AND METHODS: 74 patients hospitalized due to threatening preterm labour. 51 women gave birth prematurely; the remaining 23 were diagnosed with false labour. We used ELISA arrays to study 13 proteins: IGFBP-1, IGFBP-2, BDNF, L-Selectin, E-Selectin, ICAM-1, PECAM, VCAM-1, MIP-1 delta (MIP-1d) MIP-3ß (MIP-3b), Eotaxin-1, Eotaxin-2, BLC. RESULTS: An increased risk of preterm labour should be expected when the serum concentration for: IGFBP-1 > 158.83 pg/ml (sens. 0.608, sp. 0.609, p < 0.0001); MIP-1d < 27.66 pg/ml (sens. 0.627, sp. 0.627, p = 0.021); BDNF >36.54 pg/ml (sens. 0.630, sp. 0.647, p = 0.002); BLC >25.46 pg/ml (sens. 0.588, sp. 0.609, p < 0.001); Eotaxin-1 >1.16 pg/ml (sens. 0.633, sp. 0.652). CONCLUSION: There have been reported statistically significant differences in serum concentrations of selected proteins in women with preterm labour and false labour.

Expression of selected genes in preterm premature rupture of fetal membranes.

OBJECTIVE: To analyse the expression of 15 genes encoding receptors and enzymes associated with the molecular mechanism of the tocolytic drugs atosiban (oxytocin receptor antagonist), nifedipine (calcium channel blocker) and celecoxib (selective cyclo-oxygenase-2 inhibitor) in preterm labor patients with premature rupture of fetal membranes in relation to symptoms of intrauterine infection and preterm labor risk factors. DESIGN: Experimental molecular study. SETTING: Tertiary obstetric care center. SAMPLE: Myometrial samples were obtained during cesarean sections from 35 patients who delivered preterm with unverified symptoms of intrauterine infection, 35 patients who delivered preterm without symptoms of intrauterine infection and 90 women who delivered at term. METHODS: The Micro Fluidic Profiling Card analytic system was used to evaluate mRNA expression of the genes of interest. MAIN OUTCOME MEASURES: The relative quantification values for mRNA expression. RESULTS: The median oxytocin receptor and cyclo-oxygenase-2 mRNA expression in preterm patients with clinical symptoms of intrauterine infection was significantly higher than in preterm patients without symptoms. The median mRNA expression of ß(1) , ß(3) and ß(4) subunits of the L-type calcium channel and prostaglandin E(2) receptor was significantly higher in preterm patients compared with term patients. CONCLUSIONS: The mRNA expression of hormones, enzymes and their receptors associated with tocolytic actions can differ in various clinical conditions. The expression of these genes is regulated at different levels and can be modified by inflammatory factors, which affect their functions.

The dynamics of endometrial growth and the triple layer appearance in three different controlled ovarian hyperstimulation protocols and their influence on IVF outcomes.

The impact of endometrial growth to the triple layer, endometrial thickness, and echogenicity on IVF outcomes was investigated in the study. A retrospective analysis of 583 ICSI patients was conducted: 385 with a long GnRH agonist protocol, 114 with a short GnRH agonist, and 84 with a GnRH antagonist protocol. The progression of endometrial growth to the appearance of the triple layer, endometrial thickness, and echogenicity was compared between protocols. At least one high quality blastocyst was transferred in a double embryo transfer. The time of the appearance of the endometrial triple layer was statistically significant for the pregnancy rate only in the GnRH antagonist protocol. The endometrial thickness on the day of the appearance of the triple layer had a statistically significant influence on the pregnancy rate in the GnRH antagonist and in the long GnRH agonist protocols. The highest pregnancy rate for the long GnRH agonist and the GnRH antagonist protocols was observed when the endometrium thickness was 12-13 mm (61.6% and 58.8%, respectively). The endometrial echogenicity had a significant influence on the pregnancy rate only in the long GnRH agonist protocol. Endometrial features could be helpful parameters in IVF outcomes in particular controlled ovarian hyperstimulation protocols.

Total matrix metalloproteinase-8 serum levels in patients labouring preterm and patients with threatened preterm delivery.

Preterm labour and prematurity are still a main cause of perinatal morbidity nowadays. The aim of our study was to assess the role of MMP-8 as a predictive marker of preterm delivery. Four groups of patients were involved to the study: I - pregnant women at 24-34 weeks of gestation with any symptoms of threatened preterm labour; II - threatened preterm labour patients between 24-34 weeks of gestation; III - preterm vaginal delivery patients; IV - healthy term vaginal delivery patients. Serum concentration of total MMP-8 was measured using two enzyme-linked immunosorbent assays. There were no significant differences in the median concentrations of total MMP-8 between physiological pregnancy and threatened preterm labour patients with existing uterine contractility. No significant differences of total MMP-8 were either found between healthy term and preterm labouring patients. The studies on a larger population are needed to reject the hypothesis that preterm labour is connected with increased MMP-8 plasma concentrations of women in preterm labour and threatened preterm delivery.

Vitrification vs. slow cooling protocol using embryos cryopreserved in the 5th or 6th day after oocyte retrieval and IVF outcomes.

Modifying cryopreservation protocols may be seen as a way to simplify cryobanking procedure and increase satisfying outcomes. The aim of the study was to evaluate the influence of slow cooling protocol and vitrification on IVF outcomes using embryos preserved in the 5th or 6th day after oocyte retrieval. The study compared 2 groups of human embryos underwent slow cooling protocol (n=189) and vitrification (n=58). All embryos were cryopreserved in the 5th or 6th day after oocyte retrieval. Pre- and postfreezing embryo evaluation was performed in 2 or 3 steps scale, respectively. The study evaluates the effectiveness of two freezing methods and influence of the freezing day, pre- and postfreezing embryo grading on clinical pregnancy rate. Study showed higher pregnancy rate after vitrification (50.4%) than slow cooling protocol (25.9%). Significantly higher pregnancy rate was observed, when embryo preserved in the 5th day after oocyte retrieval (50.3%) than in the 6th day (22.7%). Postfreezing embryos evaluation showed that high quality blastocysts gave nearly four times better pregnancy outcomes than the ones evaluated as poor quality, and three times better than the ones evaluated as moderate. Prospective trials are needed to evaluate pregnancy and neonatal outcomes after vitrification. The number of controlled studies concerning vitrification has not been large enough, yet.

[Evaluation of adiponectin and TNFalpha genes expression in women with gestational diabetes. Preliminary results]. / Ocena ekspresji genów adiponektyny i TNFalpha w tkance tluszczowej i lozysku kobiet z cukrzyca ciazowa, badania wstepne.

OBJECTIVES: Recent studies suggest the essential role of different cytokines realised from adipose tissue in pathogenesis of gestational diabetes. The aim of the study was evaluation of adiponectin (diabetes development protective factor) and TNFalpha (one of the most important insulin resistance mediator) genes expression in maternal visceral and subcutaneous adipose tissue as well as placental tissue. MATERIAL AND METHODS: The study group consists of patients with gestational diabetes, healthy pregnant glucose tolerant women represented the control group. Tissue samples--placental tissue, visceral and subcutaneous adipose tissue--were obtained from patients who were undergoing ceasarean section. RT-PCR technique was performed for evaluation of adoponectin and TNF alpha genes expression. RESULTS: Our study showed decreased adiponectin gene expression and increased TNFalpha gene expression in visceral adipose tissue in pregnant women with gestational diabetes. An expression of adiponectin gene in diabetic placental tissue was not observed. We also noticed slight increase of TNF alpha gene expressionin placenta of diabetic cases. CONCLUSIONS: Decreased adiponectin and increased TNFalpha genes expression in adipose tissue of pregnant women with gestational diabetes seems to play a significant role in insulin resistance appearance and can lead to development of diabetes in pregnancy.

[Angiogenesis-prognostic factor in patients with endometrial cancer]. / Angiogeneza-czynnik prognostyczny u kobiet chorych na raka blony sluzowej trzonu macicy.

Endometrial carcinoma is one of the most commonly found cancers. In numerous kinds of cancers, tumor microvessel density correlates with clinical stage of disease and is considered as an independent prognostic factor. Evaluation of angiogenesis intensity in endometrial cancer also seems to be an independent prognostic factor and statistically correlates with FIGO stage of disease, histological type and grade of tumor, depth of myometrial invasion and metastasis. Activity of angiogenic factors in human tissues and serum provides additional reference concerning the growth and progression of endometrial cancer.