Paweł Kuć

Fetal membranes as a source of stem cells

In recent years, a constant growth of knowledge and clinical applications of stem cells have been observed. Mesenchymal stromal cells, also described as mesenchymal stem cells (MSCs) represent a particular cell type for research and therapy because of their ability to differentiate into mesodermal lineage cells. The most investigated source of MSCs is bone marrow (BM). Yet, collection of BM is an invasive procedure associated with significant discomfort to the patient. The procedure results in a relatively low number of these cells, which can decrease with donors age. Therefore, it seems to be very important to find other sources of mesenchymal stem cells nowadays. A human placenta, which is routinely discarded postpartum, in spite of its natural aging process, is still a rich source of stem cells capable to proliferate and in vitro differentiate in many directions. Besides homing and differentiation in the area of injury, MSCs there elicit strong paracrine effects stimulating the processes of repair. In this review, we focus on the biology, characteristics and potential clinical applications of cells derived from human fetal membranes: amnion and chorion.

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.

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.

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 (GROα/CXCL1, GROβ/CXCL2 and GROγ/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-3α/CCL20, MIP-3-β/CCL19, MPIF-1/CCL23, NAP-2/CXCL7, MSPα, 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-3α, 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-3β (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.

The effect of combined tocolysis on in vitro uterine contractility in preterm labour

PURPOSE Animal models have confirmed high efficiency of combined tocolytic treatment in preterm labour. In humans, the recommended doses of tocolytic drugs prolong pregnancy in threatened preterm labour. The aim of the study was to evaluate the inhibitory effect of dual combinations of atosiban, nifedipine and celecoxib on human myometrial strips contractility on the in vitro model of preterm labour. MATERIAL/METHODS Two groups of patients who delivered by cesarean section were involved in the study: 36 patients who delivered preterm between the 24(th) and 34(th) week of pregnancy and 40 patients who delivered at term. Myometrial samples were obtained from the lower uterine segment during cesarean sections. Contractile activity was recorded with digital software for each drug combination: atosiban/nifedipine; atosiban/celecoxib, nifedipine/celecoxib. Tocolytic efficiency of the drug combinations was assessed using IC(50) parameter - a molar drug concentration inhibiting 50% of contractility. RESULTS The atosiban/nifedipine combination has shown additive tocolytic effect on myometrial strips contractility in preterm and term patients. The other combinations: atosiban/celecoxib and nifedipine/celecoxib presented only antagonistic effects in both studied groups. CONCLUSIONS The effect of the combined therapy on human myometrial contractility presented in the study could be a base for further in vivo clinical trials.

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.

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 womens 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.

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

HighlightsIncrease in the concentration of 8 proteins in the plasma of women with preeclampsia.IFN‐&ggr;, IL‐6, LIF, Hb‐EGF, HGF, IP‐10, leptin, PDGF‐BB.Significant decrease in the concentration of 3 proteins in the plasma of women with preeclampsia.VEGF, PlGF and follistatin. 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‐&ggr;), 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.

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.