LC/MS/MS assessment of changes in placental angiotensin I metabolism in preeclampsia.

BACKGROUND: Preeclampsia (PE) is a condition characterized by high blood pressure and significant proteinuria in pregnant women. It affects about 7% pregnancies and can be cause of fetal and maternal morbidity and mortality. During pregnancy, a physiological overexpression of the Renin-Angiotensin System (RAS) components is observed, including increased plasma Ang II level. Dysregulation of RAS in placenta may contribute to preeclampsia and uterine growth retardation. The aim of the study was to evaluate the Ang I metabolism in human preeclamptic placentas and to compare to normal pregnancies condition. METHOD: Fragments of placental tissues were collected right after ceasarian section from PE and physiological pregnancies. Tissues were incubated in Krebs buffer in the presence of Ang I. Evaluation of Ang I metabolites in incubating fluid was performed by LC/MS/MS method. mRNA expression of main RAS components was measured by RT-PCR. RESULTS: Pattern of angiotensin metabolites did not differ between groups. The main products were Ang 1-7 and Ang II. Comparing to control group, more than 3-fold lower production of Ang II and Ang 1-7 in preeclampsia was observed. mRNA expressions of ACE and AT1 were significantly decreased in pre- eclamptic placentas, whereas higher expression of mRNA of ACE2 and MAS receptor were observed. CONCLUSIONS: Production of Ang 1-7 by PE placentas was significantly lower than in control group. Significantly decreased mRNA expression of ACE and AT1 receptor and lower production of Ang II in placentas of PE patients suggest that placental Ang II/ACE/AT1r pathway could be less important than Ang 1-7/ACE-2/MASr pathway in development of preeclampsia, but this requires further investigations.

The dynamics of autonomic nervous system activity and hemodynamic changes in pregnant women.

OBJECTIVE: The purpose of this research was to assess the dynamics of autonomic nervous system(ANS) and hemodynamic activity changes during uncomplicated pregnancy. METHODS: We enrolled 36 pregnant women (mean age 29 ± 4.8 years) and a control group of 10 non-pregnant women (mean age 25.9 ± 0.88 years). The examination was performed in the 1st, 2nd, and 3rd trimester. Continuous registration of BP, ECG, and cardioimpedance was performed with Task Force Monitor 3040i. ANS activity was measured using the following parameters: HRV, BPV, BRS at rest, and in response to autonomic tests. RESULTS: Compared to the 1st trimester, an increase in HR (73 vs. 92 bpm; p < 0.001) and mean BP (80 vs. 85 mmHg, p < 0.01) was observed in the 3rd trimester. In the 1st trimester, the BRS of pregnant women was insignificantly higher than in the controls (24.8 vs. 22.3 ms/mmHg); subsequently, it decreased significantly, to 13.4 ms/mmHg in the 3rd trimester (p = 0.0004). An increase in nLF (39.57 ± 13.75 vs. 58.73 ± 15.55; p = 0.001) and LF/HF ratio (1.03 ± 0.76 vs. 1.85 ± 0.8; p < 0.00002) was revealed in HRV analysis conducted in the 3rd trimester, as compared to the 1st tri- mester, along with a decrease in nHF (60.43 ± 13.71 vs. 41.26 ± 15.55; p < 0.001). An increase in LF/HF-sBPV (1.05 ± 0.48 vs. 1.58 ± 0.44; p = 0.01) was recorded in BPV analysis at rest in the 3rd trimester as compared to the respective 1st trimester value. CONCLUSION: Our findings suggest that pregnancy is associated with dynamic changes in autonomic balance, namely doubled dominance of the sympathetic component. Hypervolemia seems the major factor responsible for autonomic and hemodynamic changes observed during pregnancy, as it causes an increase in BP and simultaneous decrease in BRS.

Association of single nucleotide polymorphism rs7579169 with hypertension disorders during pregnancy and perinatal outcome.

BACKGROUND: Hypertension during pregnancy is a heterogeneous group of disorders with elevated blood pressure with or without proteinuria. The multiple researches are held on the subject of a genetic conditioning of preeclampsia and pregnancy induced hypertension. OBJECTIVES: The study was designed to evaluate the impact of the single nucleotide polymorphism (SNP) rs7579169 on hypertension disorders in pregnancy, especially on PE and PIH as well as on the perinatal outcome. METHODS: It is a case-control study. The study included 104 women with uncomplicated pregnancies in the control group and 75 pregnant women with hypertension disorders in the study group, hospitalized in the Perinatology and Obstetrics Department of the University Hospital in Cracow. Genomic DNA was extracted from peripheral blood leukocytes and SNP rs7579169 was genotyped from all patients. We analyzed the genotypes distribution and allele frequencies of polymorphism rs7579169 and its association with perinatal outcome in all groups. A p-value<0.05 was considered as significant. RESULTS: Clinical evaluation included standard anthropometric measures like weight and height for the calculation of the body mass index in the beginning and in the end of the pregnancy, blood pressure, time and a method of delivery, birth weight, Apgar score. The heterozygote CT was associated with a 4.5-fold increased risk of preeclampsia in pregnant patients. The presence of TT genotype significantly increased the risk of intrauterine growth restriction (<10 percentile). CONCLUSIONS: The study show probable impact of SNP rs7579169 on pregnancy, but further studies on larger groups are needed.

[Pregnancy in patients with cardiomyopathy--cases presentation]. / Ciaza u pacjentek z kardiomyopatia--opis przypadków.

It is estimated that 0.2-4% of all pregnancies are complicated by cardiovascular diseases in industrialized countries and the number of the patients with cardiac problems in pregnancy is currently increasing. Cardiomyopathy is a heart disease, congenital or acquired, that may cause heart impairment and as a consequence it may lead to insufficient adaptation under conditions of increased load during gestation. Cardiomyopathy during pregnancy occurs relatively rarely, but potentially it may have severe consequences. Moreover up to 11% of maternal deaths during pregnancy may be caused by cardiomyopathies. We present a series of clinical cases of cardiomyopathy diagnosed before pregnancy. In our observation there are two pregnant women with the hypertrophic cardiomyopathy and one pregnant woman with the dilated cardiomyopathy. Our experience corresponds with previous reports. Pregnancy in the course of cardiomyopathy, leading to changes in blood volume, cardiac output and peripheral vascular resistance, with the limited capacity of the heart, is a therapeutic challenge. The key to the successful therapy is to quantify precisely the risks for both the mother and the fetus. The counseling should be provided by a specialist with appropriate experience, and should be performed in the cooperation with obstetricians, cardiologists and anesthetists.

[Selected polymorphisms of Renin-Angiotensin System in the pathology of hypertensive disorders of pregnancy]. / Wybrane polimorfizmy genów ukladu renina-angiotensyna w patogenezie nadcisnienia w przebiegu ciazy.

Pregnancy induced hypertension and pre-eclampsia, defined as hypertension and proteinuria after 20 weeks of pregnancy with no history of such symptoms, are a great challenge in the field of perinatology They are a serious threat to the mother and the child. Regardless of numerous studies on the subject, the pathogenesis of pregnancy-induced hypertension and pre-eclampsia is still unknown, although a crucial role of genetic factors combined with environmental factors has been confirmed. New theory based on genetic analysis of Renin-Angiotensin System and its impact on blood pressure in pregnant women, has been subject to much debate recently Numerous genes have been studied but angiotensinogen remains to be the best known. Little is known about angiotensin receptor type 1 and 2. This paper presents the knowledge about selected genes of RAS and their impact on pregnancy

Assessment of angiotensin I metabolism in the human placenta using an LC/MS method.

The local renin-angiotensin system (RAS) in the placenta plays a very important role in placental development. It is well known that during normal pregnancy most of the circulating and local RAS components are over-expressed and any disruption of this new balance may cause pregnancy complications. The aim of this study was to assess the metabolism of Ang I in placentas from normal pregnancy, in an ex vivo model, using an LC/MS method. The obtained results suggest that placental tissue is able to produce many angiotensin peptides but the main metabolite is Ang-(1-7).

Leptin and interferon-gamma as possible predictors of cesarean section among women with hypertensive disorders of pregnancy.

BACKGROUND: This study was designed to evaluate maternal levels of leptin and interferon-gamma (IFN-gamma) in pregnancy complicated with hypertension and to assess the role of cytokines in predicting the risk of cesarean section. MATERIAL/METHODS: This was a cohort study with a prospective follow-up. After proportional sampling procedure, the study included the follow-up of 40 women with hypertensive disorders of pregnancy (pregnancy-induced hypertension [PIH] or preeclampsia [PE]) and 40 uncomplicated pregnancies. Women were followed from the time of admission to the delivery. Levels of leptin and interferon-gamma were measured in serum samples from all women. A p-value <0.05 was considered as significant. RESULTS: Significant increase in IFN-gamma and leptin concentration in women with pre-eclampsia was observed. We found a significant 1.4-fold increase in the risk of birth by cesarean section associated with the increase of the IFN-gamma concentration by 0.1 pg/ml and almost 3-fold increase in the risk associated with the increase of the leptin concentration. CONCLUSIONS: IFN-γ and leptin might be risk markers of cesarean section in hypertension disorders of pregnancy, but further studies supporting this evidence are needed.

Effect of pentoxifylline, administered in preterm labour, on the foetal-placental circulation and neonatal outcome: a randomized, prospective pilot study.

The aim of the study was to evaluate the pentoxifylline administration on the foetal-placental circulation and neonatal outcome in women with threatened preterm labour. Pentoxifylline was given as a supplement to standard tocolytic therapy in a group of 43 patients (pentoxifylline group) as an intravenous infusion and oral supplementation in a total dosage of 800 mg/day. The drug was administered within 3 weeks after admission. No pentoxifylline was given in the control group (53 patients). Doppler velocimetry of pulsatility indices (PI) of the umbilical (UA) and middle cerebral (MCA) arteries as well as cerebro-placental ratio (CPR) were calculated. Also, the neonatal outcome was estimated in both groups. From the second week of therapy with pentoxifylline, the PI decreased in umbilical artery and increased in the MCA, whereas in the control group, there were no changes. The value of PIUA, evaluated after the third week of pentoxifylline administration, was statistically significantly lower when compared to data obtained on admission (mean: 0.99 ± 0.22 versus 0.82 ± 0.12; p =0.016). Pentoxifylline significantly increased CPR values calculated after third week of drug administration, which were statistically significantly higher in the pentoxifylline group when compared with respective data in the control group (mean: 2.30 versus 1.61; p = 0.001). The risk of severe neonatal complications was significantly lower in the pentoxifylline group (p = 0.026). Pentoxifylline changed foetal-placental blood circulation in patients with threatened preterm labour and improved neonatal outcome.

Aspirin resistance may be associated with adverse pregnancy outcomes.

OBJECTIVE: Verify that resistance to aspirin may have an impact on pregnancy and neonatal outcome. METHODS: We enrolled 43 pregnant women, aged 30.7 ± 4.0 years regularly taking 75 mg of aspirin daily and 32 (aged 30.8 ± 4 years) pregnant women not receiving aspirin who served as control group. Laboratory tests were performed at 18 to 22 weeks of gestation, 28 to 32 weeks of gestation and 16 to 32 weeks after delivery. Resistance to aspirin was defined as urinary 11-dehydrothromboxane B2 (u11-dTXB2) concentrations in the highest quartile and additionally, as the resistance index (RI) calculated for each woman, defined as the difference between u11-dTXB2 concentration of each woman treated with aspirin and the median value at the same time point measured in the control group. RESULTS: Women taking aspirin in the highest quartile of u11-dTXB2 delivered prematurely (35.8±3.4 vs 38.1±1.7 weeks, p=0.02). Delivery of small for gestational age (SGA) newborns (p=0.003) as well as fetal distress (p=0.014) and preeclampsia (p=0.003) occured more frequently in aspirin-resistant women. Resistance to aspirin based on the RI value was also associated with higher prevalence of preeclampsia (p=0.02) and SGA newborns delivery (p=0.01). The two groups resistant to ASA designed on the basis of both (RI and u11-dTXB2 urine levels) methods compared with ASA sensitive group differed in frequency of SLE prevalence. CONCLUSION: Aspirin resistance may be associated with increased risk of adverse pregnancy outcomes including preeclampsia, premature delivery and delivery of SGA newborns.

Doppler velocimetry of the materno-fetal circulation in preterm delivered pregnancies complicated with hypertension.

BACKGROUND: Hypertension is one of the most frequent complications of pregnancy. Due to high risk of morbidity and mortality in both mothers and children, it is necessary to continuously monitor the pregnancy, principally with biophysical methods. Particularly, doppler velocimetry of the materno-fetal circulation proves useful. THE AIM of the study was to assess the usefulness of doppler test in monitoring the condition of the foetus in preterm delivered pregnancy complicated with hypertension. MATERIAL AND METHODS: The retrospective analysis comprised the data of 116 women who delivered prematurely at the Clinics of the Department of Gynaecology and Obstetrics at the Collegium Medicum of the Jagiellonian University in the years 2006-2007, resulting in creation of Group I involving 38 pregnant women with preeclampsia, and Group II of 36 women whose pregnancy was complicated with gestational hypertension. Control group was formed of 42 women with correct arterial blood pressure. When describing the groups, the differences in the birth weight and Apgar score were indicated. RESULTS: A significant statistical difference was found in the area of pulsation rate in the umbilical artery and cerebro-placental ratio (CPR). In the case of preterm delivery complicated with arterial pressure disorders, the foetus is characterised with worse organic perfusion and slower somatic growth than if no concomitant hypertension is present. Hypertension forms an additional risk factor in the course of preterm delivery, and doppler velocimetry is a good method for monitoring the condition of the foetus, as it allows for detection of irregularities and for implementation of relevant treatment to improve the newborn's condition at birth.

Changes in the subpopulation of CD25+ CD4+ and FOXP3+ regulatory T cells in decidua with respect to the progression of labor at term and the lack of analogical changes in the subpopulation of suppressive B7-H4 macrophages--a preliminary report.

PROBLEM: The initiation of labor is accompanied by alterations in the level of maternal immune tolerance toward fetal antigens. It is a complex molecular response leading to a brief activation of the maternal immune system with an accompanying capacity to restrict this same activation. The aim of our study was to evaluate the subpopulation of regulatory T cells (Tregs) and B7-H4 macrophages in the decidua basalis during cesarean sections performed on patients in various stages of labor. METHOD OF STUDY: The decidual tissue samples evaluated in our study were obtained from 23 pregnant women who underwent cesarean sections at term. Moreover, the patients were divided into three subgroups according to the progression of labor at the time of the cesarean. The presence of Treg cells and B7-H4 positive macrophages were analysed by fluorescence-activated cell sorter. RESULTS: The percentages of FOXP3+ cells in the subpopulation of CD25+ CD4+ T lymphocytes found in the deciduas of patients decreased with the successive stages of labor, while the percentages of B7-H4 positive cells in the macrophage subpopulation remained almost constant. CONCLUSION: These changes in the Treg cell subpopulation in the decidua would seem to be related to a brief activation of the maternal immune system as labor begins and lack of analogical changes in the subpopulation of decidual suppressive B7-H4+ macrophages that enable the restriction of this same activation as labor progresses.

Group B streptococcus colonization of pregnant women and their children observed on obstetric and neonatal wards of the University Hospital in Krakow, Poland.

The study was arranged to assess the actual rates of colonization of pregnant women and their children with group B streptococcus (GBS) in a Polish university hospital. Resistance of these cocci to macrolides and clindamycin was also tested and routes of transmission of GBS were followed in some cases using molecular typing. Colonization with GBS was checked in 340 pregnant women living in the south-eastern region of Poland (Malopolska) in the years 2004-2006. Women with a complicated pregnancy were more often colonized than those with a normal pregnancy (20.0 % versus 17.2 %). Moreover, women with a complicated pregnancy were twice as often colonized with GBS strains with the MLS(B) phenotype indicating resistance to macrolides and clindamycin. Regarding neonatal colonization by GBS, we found that neonates born from the colonized mothers with a complicated pregnancy were more often colonized with GBS than those from the mothers with a normal pregnancy (35 % versus 26.7 %). By molecular typing of the GBS strains isolated from mothers and their newborns we have been able to suggest the possibility of horizontal transmission of the strains from the hospital environment to newborns. Our results clearly indicate that rates of GBS colonization among pregnant women and neonates in a Polish university hospital have reached levels comparable to those reported in other European clinical centres.

[Feto-placental blood flow--practical remarks]. / Zaburzenia przeplywu lozyskowego--praktyczne zastosowanie kliniczne.

Pregnancy is a physiological state which still needs close observation from the very beginning till the delivery to decrease the potential risk of complications for the mother and her baby. The finding of pathology is possible thanks to advanced diagnostic technology--cardiotocograph and ultrasonography, especially Doppler technique. Monitoring of the fetus is based not only on the routine obstetric examination, but also on the observation and analysis of the fetal heart rate, biometry measurements with expected fetus weight, as well as assessment of blood flow in the fetal, the umbilical and uterine vessels. Assuming that pathology in fetal heart rate can predict intrauterine discomfort, early examination and appropriate reaction to the situation is essential. Early identification of intrauterine growth restriction enables intensive treatment which improves the outcome and reduces perinatal mortality. Information concerning biophysical monitoring of a fetus and basic rules of test result interpretations of are presented in this article.

[Physiological maternal changes during pregnancy and their consequence in GP-practice]. / Zmiany fizjologiczne w organizmie kobiety ciezarnej i ich znaczenie w praktyce lekarza ogólnego.

Physiological maternal changes, as well as signs and symptoms during pregnancy were presented. Changes in cardiopulmonary, urinary, digestive tract, metabolic and endocrinal changes in the light of practical application in G-P practice were presented.

[Increasing Streptococcus agalactiae colonization of pregnant women and newborns in south-eastern region of Poland]. / Narastanie stopnia kolonizacji kobiet w ciazy i noworodków przez Streptococcus agalactiae na obszarze Polski poludniowo-wschodniej.

Streptococcus agalactiae, group B streptococci (GBS) are a constituent of normal vaginal bacterial microflora which often do not give any clinical symptoms. On the other hand, during pregnancy there are optimal conditions for GBS multiplication in the vagina, which may have very serious consequences for both the mother and her child. The women (n = 563) that participated in our study were in their 3rd trimester and they were divided into groups: normal pregnancy or high risk pregnancy. We also examined their newborns. GBS identification was done basing on traditional culture method and its modification recommended by the CDC. We showed a slightly improved (about 4%) effectiveness of GBS detection in pregnant women using the CDC method. In high risk pregnancy GBS colonization was 20% (among them 35% newborns were colonized) and in normal pregnancy it was found to be 17.2% (among them 26.7% newborns were colonized). Both in the high risk group and their newborns we confirmed a higher and statistically significant frequency of detection of GBS strains which had MLS(B) mechanism of antibiotic resistance. In newborns we confirmed two cases which were fatal. The results of our study show the need and necessity for implementing unified procedures recommended by the CDC in Poland.

Effects of oral L-arginine on the pulsatility indices of umbilical artery and middle cerebral artery in preterm labor.

OBJECTIVE: The objective of the study was the estimation of the influence of oral supplementation with low-dose l-arginine on feto-placental circulation in women with threatened preterm labor. STUDY DESIGN: Oral administration of 3g of L-arginine daily or placebo as a supplement to standard tocolytic therapy was tried in 70 women with threatened preterm delivery, randomly assigned to the L-arginine (n=37) or placebo (n=33) groups. Twenty-five and 20 completed the study, respectively. Doppler velocimetry of pulsatility indices (PI) of the umbilical (UA) and middle cerebral (MCA) arteries as well as pregnancy outcome and biochemical markers of nitric oxide synthesis (plasma amino acid and nitrite/nitrate levels, as well as 24 h nitrite/nitrate excretion with urine) were estimated. RESULTS: Starting from the second week of therapy, the UA PI values were significantly lower in the L-arginine group than in the placebo group. Moreover, treatment with L-arginine caused a significant increase in MCA PI and cerebro-placental ratio (CPR) values. The changes in feto-placental circulation in the L-arginine group were not associated with any signs of increased nitric oxide synthesis. CONCLUSION: Oral supplementation with low doses of L-arginine changed feto-placental blood flow distribution in patients with threatened preterm labor. The exact mechanism of L-arginine action on feto-placental circulation requires further investigation.

RCAS1 decidual immunoreactivity in severe pre-eclampsia: immune cell presence and activity.

PROBLEM: Pre-eclampsia seems to be related to the disturbance of immune tolerance regulation during pregnancy. Receptor-binding cancer antigen expressed on SiSo cells (RCAS1) decidual level alterations were concomitant with changes in immune cell number and activity in decidua. As decidual immunomodulating activity participates in the development of immune tolerance during pregnancy, we aimed to evaluate the immunoreactivity level of decidual RCAS1 with respect to the presence and activity of immune cells. METHOD OF STUDY: RCAS1, CD3, CD56, CD69, and CD25 immunoreactivity was assessed by immunohistochemistry in 30 decidual samples derived from patients with severe pre-eclampsia (sPE) and from a healthy control group. RESULTS: RCAS1 immunoreactivity was statistically significantly higher in decidual tissue samples derived from patients with sPE tissue than in those derived from healthy patients in whom elective cesarean section at term was performed. A statistically significantly lower number of CD56(+) and CD3(+) cells and lower immunoreactivity level of CD69 were found in patients with sPE compared with those from the control group. CONCLUSION: The limited immune cells infiltration in decidua during sPE is associated with increase in RCAS1 decidual level.

Nebulized pentoxifylline for prevention of bronchopulmonary dysplasia in very low birth weight infants: a pilot clinical study.

OBJECTIVE: To evaluate the effectiveness of nebulized pentoxifylline (PTXF) compared to intravenous dexamethasone (DX) or placebo (nebulized distilled water) for the prevention of bronchopulmonary dysplasia (BPD). METHODS: One hundred and fifty very low birth weight infants were randomly assigned to three groups. Entry criteria were the need for oxygen administration on the fourth day of life, irrespective of whether ventilatory support was required. PTXF was administered with a nebulizer every 6 hours on three consecutive days (a single course) in a dose of 20 mg/kg when infants were breathing spontaneously or 10 mg/kg when they needed ventilatory support. DX was given every 12 hours on three consecutive days in a dose of 0.25 mg/kg. Nebulized distilled water was administered with the schedule of inhalation as in the PTXF group. When the need for ventilatory support or oxygen dependency persisted, the course of both drugs and placebo administration was repeated every seven days until the diagnosis of BPD was established. RESULTS: Both PTXF and DX reduced the incidence of disease when compared with placebo. The respective data obtained for the PTXF-group versus the placebo group were as follows: difference in risk, 27%; OR: 0.32; CI: 0.11-0.94; p = 0.039; whereas the results for the DX-group versus the placebo group were: difference in risk, - 23%; OR: 0.39; CI: 0.14-1.14; p = 0.07. CONCLUSION: Our data show that nebulized PTXF reduces the risk of BPD and may be a potential alternative to steroids in the prevention of this disease.

Effects of oral L-arginine on the foetal condition and neonatal outcome in preeclampsia: a preliminary report.

Estimation of the influence of oral supplementation with low dose of L-arginine on biophysical profile, foeto-placental circulation and neonatal outcome in preeclampsia. Randomized, placebo-controlled, double-blind, clinical trial. Oral therapy with 3 g of L-arginine daily or placebo as a supplement to standard therapy. Eighty-three preeclamptic women, randomly assigned to the L-arginine (n=42) or placebo (n=41) groups; [n=30 (L-arginine) and n=31 (placebo) ended the study, respectively]. Foetal gain chances due to ultrasound biometry, biophysical profile, Doppler velocimetry of pulsatility indices of umbilical and middle cerebral arteries, cerebro-placental ratio, as well as differences in duration of pregnancy and clinical data of newborn. L-arginine treatment transitory accelerated foetal gain and improved biophysical profile. Starting from 3rd week of therapy, the umbilical artery pulsatility indices values were significantly lower in L-arginine than in placebo group. Moreover, treatment with L-arginine caused significant increase of middle cerebral artery pulsatility indices and cerebro-placental ratio values. Latency was longer in L-arginine group. Neonates delivered in the L-arginine group revealed higher Apgar score. Supplementary treatment with oral L-arginine seems to be promising in improving foetal well-being and neonatal outcome as well as in prolonging pregnancy complicated with preeclampsia. However, these benefits require confirmation in more-powered, larger studies.