Role of sphingolipids in the pathogenesis of intrahepatic cholestasis.

BACKGROUND & AIMS: Intrahepatic cholestasis of pregnancy (ICP) is the most common pregnancy-related liver disorder that affects from 0.2% to 15.6% pregnant women. The disease is connected with increased risk of fetal morbidity and mortality, but is unfortunately detected quite late. The diagnosis of ICP is based on only one manifestation: pruritus which mainly affects soles and palms. METHODS: Twenty intrahepatic cholestasis of pregnancy (ICP) women and twenty healthy pregnant women (control group) took part in the study. In the study group, blood sampling for baseline measurements was performed on the first day of hospital stay - before the commencement of treatment with ursodeoxycholic acid (UDCA) - and repeated after 7 days of 900 mg UDCA per day. An additional blood sample was collected on the second day after childbirth. In the control group, blood samples were collected directly after hospital admission. We compared plasma sphingolipids in samples of the subjects from ICP and ICP + UDCA-treated groups as well as the ICP group after delivery with the healthy controls. RESULTS: Of all sphingolipids, the median values of C16-Cer and C18-Cer were significantly higher in the plasma of cholestasis patients not treated with UDCA as compared to the control. Following 7 days of UDCA treatment, a considerable decrease in C16-Cer, C18-Cer and the total concentration of bile acids was noted as compared to the baseline. CONCLUSION: It is known that sphingolipids serve as modulators of liver regeneration. We assume these substances could be potential markers for detecting early onsets of intrahepatic cholestasis of pregnancy.

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.

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.

The expression of suppressor of cytokine signaling 1 and 3 in fat and placental tissue from women with gestational diabetes.

The suppressor of cytokine signaling (SOCS) proteins are feedback inhibitors of signaling pathways induced by cytokines, hormones and growth factors. In the present study we measured the expression of SOCS1, SOCS3, interleukin-6 (IL-6), IL-6 receptor, IL-8 and leptin mRNA in paired samples of subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and placental tissue obtained from 18 pregnant women with normal glucose tolerance (NGT) and 20 subjects with gestational diabetes mellitus (GDM), using quantitative RT-PCR. The patients with GDM had significantly higher IL-8 mRNA expression in VAT than the women with NGT (p = 0.007), whereas the expression of SOCS1, SOCS3 and other genes study did not differ significantly between the two groups. Stepwise regression analysis revealed that SOCS1 mRNA expression in VAT was significantly associated with prepregnancy BMI (ß = -0.68, p = 0.03) and IL-8 mRNA expression (ß = 0.66, p = 0.03), whereas SOCS3 mRNA expression in VAT was independently predicted by IL-6 mRNA expression (ß = 0.94, p = 0.0002, R(2) = 0.88). In conclusion, our results did not show significant differences in SOCS1 and SOCS3 mRNA expression in adipose and placental tissue obtained from pregnant women with and without GDM.

Spontaneous rupture of unscarred uterus in the early second trimester: a case report of placenta percreta.

BACKGROUND: Prevalence of uterine rupture at delivery has been recently estimated at less than 1 in 2500 deliveries. Spontaneous uterine rupture in the early mid-trimester (16 weeks gestation or less), is far less frequent. We report a case of uterine rupture due to placenta percreta in otherwise uncomplicated pregnancy CASE: A 35-year-old, gravida 5, para 5, at 15wk 2d gestation (menstrual age) with negative history of uterine scarring suddenly developed symptoms of incipient hypovolemic shock while being hospitalized for imminent miscarriage. On exploratory laparotomy we found a midline uterine rupture infiltrated by the placenta. Supracervical hysterectomy was performed. Postoperative lab analysis confirmed the elevated serum AFP levels. CONCLUSION: Abnormal placentation and subsequent uterine rupture should be taken into consideration also in women in the second trimester who have no history of uterine instrumentation.

Plasma Levels and Diagnostic Utility of VEGF in a Three-Year Follow-Up of Patients with Breast Cancer.

Breast cancer is the most common malignancy in women globally. The increasing worldwide incidence of this type of cancer illustrates the challenge it represents for healthcare providers. Therefore, new tumor markers are constantly being sought. The aim of this study was to assess plasma concentrations and the diagnostic power of VEGF in 100 patients with early-stage breast cancer, both before and after surgical treatment and during a three-year follow-up. The control groups included 50 subjects with benign breast tumors (fibroadenoma) and 50 healthy women. The VEGF concentration was determined using enzyme-linked immunosorbent assay (ELISA) and the CA 15-3 concentration was determined by chemiluminescent microparticle immunoassay (CMIA). We observed significantly higher preoperative plasma concentrations of VEGF and CA 15-3 in patients with breast cancer. VEGF, similar to CA 15-3, demonstrated high diagnostic utility in the assessment of the long-term efficacy of surgical removal of the tumor. Determinations of VEGF had the highest diagnostic usefulness in the detection of breast cancer recurrence (SE 40%, SP 92%, PPV 67%, NPV 79%). Additionally, the highest values of SE, NPV and AUC were observed during the combined analysis with CA 15-3 (60%; 84%; 0.7074, respectively). Our study suggests a promising diagnostic utility of VEGF in the early stages of breast cancer and in the evaluation of the efficacy of the surgical treatment of breast cancer as well as the detection of breast cancer recurrence, particularly in a combined analysis with CA 15-3 as a new diagnostic panel.

Decreased serum level of macrophage inflammatory chemokine-3beta/CCL19 in preterm labor and delivery.

OBJECTIVE: Chemokines are small soluble molecules which mediate leukocyte migration and may be involved in the pathophysiology of preterm labor. We aimed to determine if serum concentrations of selected chemokines are changed in preterm labor and delivery. STUDY DESIGN: A novel array-based enzyme-linked immunosorbent assay was used to quantitate serum levels of nine chemokines from a single sample: MDC/CCL22, TARC/CCL17, ITAC/CXCL11, I-309/CCL1, IP-10/CXCL10, MIP-1alpha/CCL3, -1beta/CCL4, -3alpha/CCL20 and -3beta/CCL19. Women in preterm labor who delivered (n = 17), women at preterm pregnancy not in labor (n = 13) and women in labor at term (n = 8) participated. RESULTS: In the preterm delivery group of patients, the MIP-3beta/CCL19 concentration was in mean (+/-S.D.) 70.4+/-31.7 pg/mL, which was significantly lower than that in preterm gravidas not in labor of 123+/-34 pg/mL (p < 0.001) and those in labor at term of 118+/-25.6 pg/mL (p < 0.01). The other measured chemokines did not differ significantly. CONCLUSIONS: Of a small number of examined chemokines, we were able to show that one of them, MIP-3beta/CCL19 was significantly lower in women with preterm labor and delivery. Whether or not this chemokine has a potential as biochemical marker of preterm delivery remains to be determined.

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.

Brief Communication: Maternal Plasma Autoantibodies Screening in Fetal Down Syndrome.

Imbalance in the metabolites levels which can potentially be related to certain fetal chromosomal abnormalities can stimulate mother's immune response to produce autoantibodies directed against proteins. The aim of the study was to determine the concentration of 9000 autoantibodies in maternal plasma to detect fetal Down syndrome. Method. We performed 190 amniocenteses and found 10 patients with confirmed fetal Down syndrome (15th-18th weeks of gestation). For the purpose of our control we chose 11 women without confirmed chromosomal aberration. To assess the expression of autoantibodies in the blood plasma, we used a protein microarray, which allows for simultaneous determination of 9000 proteins per sample. Results. We revealed 213 statistically significant autoantibodies, whose expression decreased or increased in the study group with fetal Down syndrome. The second step was to create a classifier of Down syndrome pregnancy, which includes 14 antibodies. The predictive value of the classifier (specificity and sensitivity) is 100%, classification errors, 0%, cross-validation errors, 0%. Conclusion. Our findings suggest that the autoantibodies may play a role in the pathophysiology of Down syndrome pregnancy. Defining their potential as biochemical markers of Down syndrome pregnancy requires further investigation on larger group of patients.

Antenatal corticosteroid therapy: a comparative study of dexamethasone and betamethasone effects on fetal Doppler flow velocity waveforms.

OBJECTIVE: To compare the effects of antenatal administration of dexamethasone and betamethasone, used in two different regimens, on fetal Doppler flow velocities. STUDY DESIGN: Sixty-seven women at risk for preterm delivery received course of corticosteroids by means of a computer-generated randomization table. The Doppler examination of the pulsatility index (PI) of the umbilical artery (UA), the middle cerebral artery (MCA) and the middle cerebral artery/umbilical artery PI ratio (MCA PI/UA PI) were performed before treatment, 24 and 72 h after the first dose of corticosteroids. The SAS system was used to perform statistical analysis. RESULTS: No significant change was observed in UA PI through dexamethasone therapy. In MCA there was a significant decrease in PI at 72 h (2+/-0.43 before and 1.68+/-0.31 after, p=0.0001). Similarly a significant decrease in MCA PI/UA PI ratio was noted (2.09+/-0.51 before and 1.83+/-0.4 after, p=0.0137). No significant changes were observed in UA PI, MCA PI and MCA PI/UA PI ratio during betamethasone treatment. CONCLUSIONS: Our results indicate significant decrease in fetal middle cerebral artery impedance at 72 h after maternal administration of the first dose of dexamethasone. Effects of dexamethasone on fetal brain warrants further research.

Inhibitory effect of barusiban and atosiban on oxytocin-induced contractions of myometrium from preterm and term pregnant women.

BACKGROUND: A synthetic oxytocin analogue, barusiban, was shown to potently inhibit oxytocin-induced activity of myometrium from term pregnant women. The responsiveness to vasopressin was not influenced by the compound. OBJECTIVE: To test the effect of barusiban and a reference compound, atosiban, on oxytocin-induced activity of myometrium from women at preterm pregnancy in comparison to myometrium from women at term. METHODS: Fifteen preterm (30-36 gestational weeks) and 12 term pregnant women (38-41 weeks) who underwent cesarean delivery donated myometrial tissue for the study. Concentration-response curves following oxytocin administration to isolated myometrial strips were recorded in control experiments, in the presence of barusiban at concentrations of 2.5, 25, and 250 nM, and of atosiban at concentrations of 25, 250, and 750 nM. Effective concentration 50% (EC50) and pA2 values were calculated. RESULTS: Both antagonists in higher concentrations increased the EC50 values to oxytocin. The median pA2 value for preterm myometrium with barusiban was 9.76 and with atosiban 7.86. For term myometrium the corresponding pA2 results were 9.89 and 7.81, respectively. None of these pA2 values differed to any statistically significant degree. CONCLUSION: The selective oxytocin antagonist, barusiban, concentration-dependently inhibits oxytocin-induced myometrial contractions of both preterm and term myometrium at least as potently as atosiban. It remains to be determined if the selectivity of barusiban for the oxytocin receptor confers an advantage over atosiban as a tocolytic in preterm labor.

Misoprostol and dinoprostone therapy for labor induction: a Doppler comparison of uterine and fetal hemodynamic effects.

OBJECTIVE: To compare the effect of misoprostol (PGE(1)) versus dinoprostone (PGE(2)) on blood flow in uteroplacental circulation during labor induction. STUDY DESIGN: Eighty-four women with indications for induction of labor were assigned to receive either misoprostol 50 microg per vagina every 4 h as needed or 0.5 mg doses of dinoprostone given intra-cervically every 6 h by means of a randomization table generated by computer. Doppler velocimetry of umbilical, uterine and arcuate arteries was performed immediately before and 2-3 h after the administration of misoprostol or dinoprostone. The SAS system was used to perform statistical analysis. RESULTS: There were no significant changes of pulsatility index (PI), resistance index (RI) and systolic/diastolic (S/D) ratio in umbilical arteries after both prostaglandin compounds. Vaginal application of misoprostol significantly increased all ratios in arcuate artery and S/D ratio in uterine artery. Intra-cervically dinoprostone significantly increased PI, RI and S/D ratio in arcuate and uterine arteries. CONCLUSIONS: Our results indicate that vaginal misoprostol and cervical dinoprostone administration increases uteroplacental resistance but does not affect umbilical blood flow. Misoprostol would be as safe and effective agent as dinoprostone for cervical ripening and labor induction.

[Evaluation of serum sICAM-1 and sVCAM-1 concentration in patient at risk of preterm delivery]. / Ocena stezen czastek adhezyjnych sICAM-1 i sVCAM-1 w surowicy krwi pacjentek z zagrazajacym porodem przedwczesnym.

Aim of the study was to assess the serum adhesion molecules concentration in patients at risk of preterm labour. Sixty-three patients were divided into two groups basing on cervical evaluation (Bishop's score > or = 6 and < 6). The assessment of sICAM-1 and sVCAM-1 levels in serum were carried out by immunoenzymatic ELISA test utilizing monoclonal antibodies (moAb) against human sICAM-1 and sVCAM-1 (ELISA system, R&Q, USA). Among the patients at risk of preterm labour and Bishop's score > or = 6 statistically higher serum levels of sICAM-1 were found. There were no statistically significant differences in sVCAM-1 levels between examined groups. We conclude that serum sICAM-1 determination might improve the prediction of preterm birth.

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.

The expression of transcription factor 7-like 2 (TCF7L2) in fat and placental tissue from women with gestational diabetes.

In the present study we showed that the expression of transcription factor 7-like 2 (TCF7L2) mRNA in visceral adipose tissue obtained from 20 women with gestational diabetes was lower than in 18 pregnant women with normal glucose tolerance (p = 0.02), however after adjusting for BMI values, the difference was not significant.

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.

Permeability of human skin to selected anions and cations--in vitro studies.

Among the many factors influencing the human body every day, metal cations and anions are some of the most important and are constantly present in atmospheric fallout. Some of them such as sulfate or nitrate anions along with different detergents may cause pathological changes within the skin. The aim of this study was to estimate whether sulfate (SO4(2-)) and nitrate anions (NO3-) as well as metal cations, magnesium (Mg2+) and calcium (Ca2+) are able to pass through isolated patches of human skin and to investigate the dynamics of ion diffusion. Concentrations of the substances corresponded to those observed in the atmospheric fallout in two regions of Poland with low (Bialowieza) and high (Katowice) pollution rate. Ion chromatography method was used to determine cations and anions levels. In vitro, we found human skin to be permeable to magnesium and calcium ions as well as sulfate and nitrate anions at concentrations observed in the atmospheric fallout in two regions of Poland having different levels of pollution. Diverse influence of sodium lauryl sulfate (SLS) detergent on the grade of metal ion permeability (acceleration in the first 10 hours of experiment) and anion permeability (lack of SO4(2-) influence) suggest the presence of different routes of ions penetration through the skin. It is also supported by different transportation characteristics of individual ions through the skin over time.