Francisco Lazaro Pereira de Sousa

Oncostatin M and leukaemia inhibitory factor trigger signal transducer and activator of transcription 3 and extracellular signal-regulated kinase 1/2 pathways but result in heterogeneous cellular responses in trophoblast cells

Leukaemia inhibitory factor (LIF) and oncostatin M (OSM) are pleiotropic cytokines present at the implantation site that are important for the normal development of human pregnancy. These cytokines share the cell membrane receptor subunit gp130, resulting in similar functions. The aim of this study was to compare the response to LIF and OSM in several trophoblast models with particular regard to intracellular mechanisms and invasion. Four trophoblast cell lines with different characteristics were used: HTR-8/SVneo, JEG-3, ACH-3P and AC1-M59 cells. Cells were incubated with LIF, OSM (both at 10ngmL(-1)) and the signal transducer and activator of transcription (STAT) 3 inhibitor S3I-201 (200µM). Expression and phosphorylation of STAT3 (tyr(705)) and extracellular regulated kinase (ERK) 1/2 (thr(202/204)) and the STAT3 DNA-binding capacity were analysed by Western blotting and DNA-binding assays, respectively. Cell viability and invasiveness were assessed by the methylthiazole tetrazolium salt (MTS) and Matrigel assays. Enzymatic activity of matrix metalloproteinase (MMP)-2 and MMP-9 was investigated by zymography. OSM and LIF triggered phosphorylation of STAT3 and ERK1/2, followed by a significant increase in STAT3 DNA-binding activity in all tested cell lines. Stimulation with LIF but not OSM significantly enhanced invasion of ACH-3P and JEG-3 cells, but not HTR-8/SVneo or AC1-M59 cells. Similarly, STAT3 inhibition significantly decreased the invasiveness of only ACH-3P and JEG-3 cells concomitant with decreases in secreted MMP-2 and MMP-9. OSM shares with LIF the capacity to activate ERK1/2 and STAT3 pathways in all cell lines tested, but their resulting effects are dependent on cell type. This suggests that LIF and OSM may partially substitute for each other in case of deficiencies or therapeutic interventions.

Estudo da morfologia vascular do leito placentário na hipertensão arterial crônica

OBJECTIVES To analyze histopathological patterns of placental bed arteries in pregnancies complicated by chronic arterial hypertension. Alterations were considered according to clinical classification of the hypertensive disorders as mild (MG); moderate (MoG) and severe (SG) for comparison with uncomplicated pregnancies, control group (CG). METHODS Placental bed biopsy was performed in 60 pregnant women; the study group was comprised of pregnant women with hypertension, subdivided in 13 with severe chronic hypertension (CH), 11 with moderate CH and 11 with mild CH, and results were compared to 25 placental bed biopsies from uncomplicated pregnancies. All the pregnant women had a gestational age of at least 28 weeks of gestation with a live fetus and were submitted to cesarean section. Hypertension was considered mild with diastolic blood pressure (DBP) 90 I? 100 mmHg, moderate DBP 100 I? 110 mmHg and severe DBP=110 mmHg. Placental bed variables selected for histological analysis were: unaltered patterns, physiological changes, medial layer disorganization, medial and intimal hyperplasic changes, acute necrosis and atherosis. RESULTS In cases with SG and MoG there was predominance of abnormal histophysiological findings: medial layer disorganization and hyperplasic changes, with a statistically significant difference when compared to MG and CG. Alteration in the medial layer was observed in these cases. The normal pattern, unaltered patterns and physiologic changes were more frequent in CG and MG. Physiological changes were the most usual finding, further, there was no acute necrosis or atherosis. CONCLUSION 1. Abnormal histophysiological findings were predominant in hypertensive pregnant women compared to the normotensive ones; 2. These patterns were more frequent, according to the severity of the hypertensive disorders: Severe, Moderate and Mild; 3. More significant abnormal findings were a change in the medial layer, mainly in the group with severe hypertensive disorders; 4. The groups with moderate and severe hypertensive disorders when compared had similar results; 5. The normal histological pattern was higher in the control group and patients with mild hypertensive disorders and disclosed a similarity between them 6. The most usual pattern was physiologic changes, which was more frequent in the control group.

PP044. Profile of pregnant women with pre-eclampsia treated in priority area

INTRODUCTION The maternity mortality rate in the Santos lowlands region, priority region of São Paulo/Brazil, is higher when compared to regions in the state of São Paulo. The hypertensive disorders are implicated in approximately 20% of death. Severe preeclampsia (PE) is recognized as the frequent diagnosis associated with maternal and fetal ominous outcomes. The recognition of potential risk support antenatal intervention which could anticipate the installation of catastrophic results. Identifying the epidemiological profile of women who may present with this evolution can guide public health policies and actions of the professionals involved in caring for these women avoiding preformed concepts and contributing to the recognition of local reality situation. OBJECTIVES Identify the epidemiological profile of women affected by severe PE. METHODS A descriptive study involving 46 pregnant women admitted to specialized hospital for high-risk pregnancy with the diagnosis of severe PE according to the criteria of NHBPEP (2000), as: blood pressure ⩾160/110mmHg, proteinuria 2.0g/24h or +2 dipstick, serum creatinine >1.2mg/dl (new onset), platelets <100,000/mm(3), microangiopathic hemolysis (increased lactate dehydrogenase), elevated alanine aminotransferase or aspartate aminotransferase, persistent headaches or other cerebral or visual disturbance, persistent epigastric pain or eclampsia in the period from January/2008 to November/2010. RESULTS The average age of the pregnant women was 25.5years (±6.5), 11 women were multiparous and nulliparous and 35 (76.1%), the average gestational age at admission was 34 4/7, developing to eclampsia and/or HELLP syndrome were 07 women (15.2%), admission often occurred before the installation of labor in (93.5%) had a previous diagnosis of chronic hypertension (30.4%) and in all cases magnesium sulfate administration with an average length of 32h (±16.6h). CONCLUSION The occurrence of severe PE was more common in nulliparous women at the age of 25years, admitted in prematurity and not in preterm labor. The outcome of seizure and/or HELLP has a considerable impact. The association of chronic hypertension with severe PE is more than 1/4 of the cases. These data are equivalent to other areas of Brazil and may contribute to emphasize the practice of prenatal acting that can recognize and pre-comment possible adverse developments and attitudes aimed at intercepting the process.

347. Study of maternal and perinatal variables in preeclampsia according to its clinical expression

Introduction The severity of the clinical expression of preeclampsia determines maternal-fetal repercussions. Recognize indicators of severity may guide the adoption of behaviors in a timely manner. Objective To relate maternal characteristics and perinatal outcomes in patients with severe preeclampsia and without signs of severity. Method Cross-sectional study. Location: Hospital Guilherme Alvaro-Santos/Brazil, January/2015-May/2016. Patients with preeclampsia (NHBPEP/2000) were divided into two groups: no signs of severity (control) and severe (study). Maternal variables: age, body mass index (BMI), parity, prenatal care, gestational age at birth, serum creatinine, aspartate aminotransferase (AST) and alanine (ALT); and neonatal: weight and admission to the Intensive Care Unit (ICU). Statistical analysis: Fisher’s exact test (p significant Results The preeclampsia group presented more commonly than women without signs of severity: age ⩾40 years (6%), obesity (56.5%) and nulliparity (38%) against 0%, 61.5%, 6.7%, respectively. Absence of prenatal care 24% against 6.7%. The childbirth occurred between 29–346/7weeks in 36% in the group with severe preeclampsia, whereas in the preeclampsia without signs of severity: 6.7%. Average weight of newborns of 3205 g and 02 (13.3%) ICU admissions, against 2528 g and 21 (42.9%). The average creatinine level was similar; AST and ALT was 18.67 and 12.53 in the control group compared to 22.63 and 15.65. Discussion Similar rates of severe preeclampsia were found in the literature. (Batista, 2009). The severe expression of preeclampsia was associated with women in the extremes of age, with previous gestation, obesity and with worse perinatal performance. Noting that these women lack counseling and qualified care. Inadequate follow-up of prenatal care may be explained by the greater number of hospital admissions in severity. It is proposed a reflection on strategies to increase vaginal parturition in this group. The variations of AST-ALT indicate hepatic involvement anterior to the renal, which will be clearer with the continuation of the study.

350. Profile of pregnant women affected by severe preeclampsia

Introduction The clinical expression of severe preeclampsia is associated to the worst maternal and neonatal outcomes. Identify the profile of women with this condition may target the adoption of care effectively. Objective Analyze the profile and major complications of pregnant women affected by severe preeclampsia. Method transversal study. Location: Complexo Hospitalar dos Estivadores/Instituto Social Hospital Alemao Oswaldo Cruz-Santos/Brazil (November/2017–March/2018). The study include 43 pregnat women admitted with the diagnosis of severe PE according to the criteria of NHBPEP (2000), as: blood pressure ⩾160/110 mmHg, proteinuria 2.0 g/24 h or +2 dipstick, serum creatinine >1.2 mg/dl (new onset), platelets Results They were identified 43 women with serious preeclampsia. Average maternal age: 29,49 years, history of previous gestations (55.81%). The prenatal had more than six appointments for 30 women (69.8%). Cessarian labor: 65,1%. Evolution to complications: HELLP syndrome (16.3%), Eclampsia (4.6%), used magnesium sulfate (93%) and admission in Intensive Care Unit (37.2%). Discussion A gravity recognized condition reaches in this priority area women in productive phase with potential socioeconomic impact, often multiparous and at least 30% not prenatal care with lots of appropriate appointmets probably for early admission. The frequency of complications and caesarean sections also leads to the need for recognition of the problem of public life, as the need for appropriate guidance and access to qualified services for it attendance.

348. Severe maternal morbidity and pathological puerperium: The impact of hypertensive syndromes

Introduction Recognizing severe maternal morbidity (MMG) allows the adoption of effective measures, with emphasis on the contribution of hypertensive syndromes (HS). Objective To analyze the characteristics and maternal-neonatal outcomes of the hypertensive syndrome selected as severe maternal morbidity and/or pathological puerperium. Method Cross-sectional study (November/2017-March/2018), location: Complexo Hospitalar dos Estivadores/Instituto Social Hospital Alemao Oswaldo Cruz-Santos/Brazil. 70 women with at least one of Santos criteria derived from the definition of MMG of the World Health Organization (2009), Waterstone et al. (2001), Mantel et al. (1998), seeking to homogenize/facilitate this definition according to clinical/laboratory/management and/or recognized as pathological puerperium: ⩾ three days of hospitalization. Variables analyzed: diagnosis of hypertension (NHBPEP/2000), age, parity, blood pressure, gestational age and way of delivery, adequate prenatal care (⩾06 appointments), maternal complications and immediate perinatal outcomes. Results We identified that 53 (75.7%) patients had HS. Average age: 28 years. Previous pregnancies: 35 (66%). Adequate prenatal care: 49 (92.4%). Cesarean section in 73.6%. Signs of severity in 62.3%, HELLP syndrome (28.3%) and eclampsia (3.8%), used magnesium sulfate (69.8%) and admission to an adult ICU (7.5%). Neonatal: 5’ Apgar score of 8–10 (96.2%), prematurity (20.8%) and neonatal ICU admission (24.5%). Discussion A high HS rate was observed among women with MMG and with prolonged hospitalization with their risks/costs. The numerically adequate number of prenatal consultations questions the quality of the care (not evaluated). There is a need to reflect on strategies that enhance vaginal delivery and reproductive planning. The frequency of complications demonstrated the severity of the clinical expression and the expressive neonatal impact: prematurity and need for ICU neonatal admission, although the Apgar score frequently above 07, reinforces the need for recognition of SH regarding its frequency and extension of postpartum complication.

349. Association between body mass index and preeclamspia

Introduction Lipotoxicity is a consequence from low grade inflammation caused by obesity. Women presenting body mass index (BMI) above 35 kg/m[b] have four times higher risk of preeclampsia development (Bianco, 1998). Objective To characterize the association of BMI and maternal and perinatal variables between puerperas with and withoutpreeclampsia. Method transversal study, Hospital Guilherme Alvaro-Santos/Brazil, (January/2015-March/2016). Antropometric data was collected from 160 women (immediate postpartum), 75 patients without preeclampsia (control group) and 85 with preeclampsia (case group). Variables: blood pressure (hospital admission), gestational age at childbirth and immediate perinatal outcome. Results Average mother age and BMI were 25 years and 26 kg/m[b] for the control group, in the group case: 29 years and 35 kg/m[b]. Patients with pre-eclampsia showed higher blood pressure: medium level of 136 mmHg (sistolyc) and 85 mmHg (diastolic). 84% of the deliveries in the case group were cesarean sections and the majority of these women had gestation resolution above 37 weeks. Newborns out of pre-eclampsia mothers weighed among 2685 g and five-minute Apgar score was 8,7. Those neonates presented higher admission rates in a ICU (36.5%) and 12% were small for gestational age. Cesarean in 84% of births in the case group and 40% case of births occurred in prematurity. Newborns of mothers with preeclampsia weighed on average 2685 g, average 5’ Apgar score of 8,7, higher rates of admission to ICU (36.5%) and 12% were small for gestational age. Discussion Was observed in women with preeclampsia higher average BMI and blood pressure values and their newborns showed smaller weight and 5’ Apgar score, need frequent admission to ICU and prematurity. Reproductive counseling appears to be relevant to the obese women to warn about potential complications, it’s important to adopt strategies for assistance to this population to qualify the attendance, particularly in actions preventive and possible reduction in the rate of cesarean section that could magnify risks.

PP181. The role of the JAK–STAT pathways and SOCS in preeclampsia

decreases insulin sensitivity and is associated with obesity. Recent data indicates that subclinical endotoxemia is associated with inflammation in obese women in late pregnancy. Objectives: The objective of this study was to quantify circulating endotoxin across pregnancy in lean and obese women, and assess the relationship between endotoxin and markers of inflammation and insulin sensitivity. Methods: Endotoxin was measured in sterile maternal EDTA plasma samples from 24 lean pregnant women (BMI = 22.4 ± 1.9 kg/m) and 45 obese pregnant women (BMI= 32.6 ± 2.1 kg/m), and 6 non-pregnant women. Samples were collected at 10.5 ± 3.1, 21.3 ± 4.6 and 35.2 ± 2.1 weeks gestation. Endotoxin was quantified using the PyroGene Recombinant Factor C endotoxin detection assay from LONZA, inter-assay variability <10%. IL-6, myloperoxidase, uric acid, triglycerides, insulin and glucose were also measured. Statistical analysis was by repeated measures ANOVA and students t-test as appropriate. Correlation analysis was performed using Pearson product moment correlation coefficient. Statistical significance was accepted at p < 0.05. Results: Endotoxin was significantly increased in both lean (10.4 ± 5.3 EU/ml) and obese (9.1 ± 5.3 EU/ml) pregnant women compared to non-pregnant women (4.3 ± 2.6 EU/ml, p < 0.05). Endotoxin increased significantly across pregnancy in both lean and obese pregnant women (p < 0.001), but was not different between these groups (table). Endotoxin was not associated with adiposity, IL-6, myloperoxidase, uric acid, triglycerides or insulin sensitivity as assessed by homeostasis model of insulin resistance (HOMA). Data are mean ± SD. Repeatedmeasures ANOVA p<0.001.

PP008. Lipotoxicity and preeclampsia: A probable correlation (preliminary results).

INTRODUCTION The impact of preeclampsia (PE) of pregnancy is seen as a global concern. Despite their importance, PE does not have well-established causes. Several studies point to obesity as a predisposing factor for PE [1]. In a systematic review Duckitt and Harrington showed that in women with BMI⩾30 the relative risk of developing PE is equal to 2.1 [2]. Bianco et al. demonstrated that the chance of patients with BMI⩾35 developing PE is more than 4 times [3]. Furthermore, it is believed that the intracellular accumulation of triglycerides cause mitochondrial dysfunction, more electron transport and generation of ROS (Reactive Oxygen Species) [4], causing a harmful effect on the endothelium. OBJECTIVES To compare the incidence of potential lipotoxicity markers among normal pregnant women and with pre-eclampsia. METHODS A case-control study, including pregnant women from October 2011 to January 2012 at Hospital Guilherme Álvaro in Santos/Brazil. The study group consisted of 27 mothers with pre-eclampsia, according to the criteria of NHBPEP (2000) and the control group consisted of 27 mothers with normal pregnancy, who gave birth vaginally, at term without hypertension. The exclusion criteria were diagnosis of collagen diseases, smoking, diabetes, twin pregnancy and fetal malformations. Blood samples were collected immediately after delivery and the results were analyzed in conjunction with maternal data, according to the following variables: age, body mass index (BMI), serum cholesterol total and fractions, glucose, triglyceride and creative protein. Analysis of SF data was performed by calculating the odds ratio adopting an hypothesis rejection level of 0.05. RESULTS The PE group had BMI>30 (or), serum triglycerides>150 (or) and LDL cholesterol>100 (or) more frequently than normal mothers, and as regards statistical significance comparing the age of the women, glucose level, HDS cholesterol, there was no significant difference between the groups. CONCLUSION In pregnancy, lipotoxicity reduces the ability of trophoblastic invasion, causing maternal systemic endothelial and placental dysfunction causing alterations in metabolic functions. All such changes maintain a close relationship with the pathophysiology of PE. Monitoring pregestational period of obese patients or those with lipid disorders is part of the medical arsenal against PE by monitoring and advising on the best time to conceive.

PP139. The elective prematurity birth and its perinatal effects of syndromes in hypertensive pregnancy

INTRODUCTION The hypertensive pregnancy disorders affect 7.5% of Brazilian women, being a major cause of maternal and perinatal morbidity and mortality, affecting 6-10% of primigravidae. They represent a significant cause of prematurity birth, with elective responsible for 20-30% of premature births, with higher incidence of neonatal morbidity due to high rates of complications in short and long term. OBJECTIVES Identify the repercussions in a short time, on caused by elective obstetric parity decisions in women with hypertensive neonatals pregnancy syndromes. METHODS Retrospective analysis of medical records of pregnant women admitted to Hospital Guilherme Álvaro, in the city of Santos/SP - Brazil, between Jan/2005 to Jan/2012. Calculations were performed for dichotomous variables and an epidemiological analysis with data obtained on pregnant women with hypertension with a single fetus, subjected to delivery before 37 weeks. Classification of Hypertensive Syndromes followed the criteria of the NHBPEP - 2000, after blood pressure measurements were calibrated following standard technique: Chronic hypertension (CH), Pre-eclampsia, superimposed pre-eclampsia and late gestational hypertension and multiple births were excluded, premature labor and fetal malformations, totaling 102 cases. RESULTS Throughout the study, it was observed that 67.6% are appropriate for gestational age, birth weight 1500-2500grams (43.1%). Support in the neonatal were excluded ICU was necessary in the vast majority (75.7%), and length of stay exceeding 28 days in 29.3% of cases. The main indication was respiratory distress syndrome (63.6%) requiring intubation orotraquel and jaundice in 18.2% of premature infants was a common comorbidity (39.4%). Even with all the support in-hospital, the neonatal mortality was present in 8.1% of cases. The main indication of the resolution was for obstetric maternal complications (61.8%) where pre-eclampsia and preeclampsia superimposed were expressions which caused more frequent (42.2 and 40.2% respectively), showing the level of proteinuria between two -4.9g (32.1%). Gestational age at delivery was 32-34 weeks (43.1%), corticosteroid therapy was not performed (54.9%) and cesarean delivery was (94.1%). The most frequent age group was 20-34 years (53.9%), Caucasian (54.9%), primigravida (62.7%) and less than five prenatal visits (36.3%) were performed. CONCLUSION The hypertensive pregnancy are risk factors for preterm elective delivery and is more frequent before 34 weeks, resulting in high perinatal morbidity and mortality, especially in cases of preeclampsia with severe clinical presentation and long hospital stays, with consequent costs and conducting excessive procedures in the neonate. Enable early referral to specialist services and consequent prenatal follow up, enables the adoption of early action, such as clinical support in a timely manner to facilitate the handling of cases which are still stable, optimizing the opportunity for administration of antenatal corticosteroids. It is necessary to review decisions about the mode of delivery in prematurity related to hypertension cases, with the aim of increasing the occurrence of vaginal deliveries.