Doctorate


Title: Effect of Pre-Gestational Obesity on the Cytokine Content of Maternal and Fetal Compartments and on the Expression of Placental Fatty Acid Transporters
Author: LIVIA BELCASTRO DE ALMEIDA
Conclusion Paper: TESE
Abbreviation: ALMEIDA, L.B.
Date of Defense: 29/08/2017
Abstract: Placental fatty acids (FA) transport proteins are responsible for a large part of the maternal-fetal transfer of these nutrients, which is fundamental for the development of the fetus. Among the factors capable of interfering with the expression of these transporters, inflammatory markers are included. Since obese individuals are prone to chronic inflammation, it is postulated that pregnant women with this condition could present changes in the amount of FA transporters in the placenta. The aim of the present study was to evaluate the concentrations of the cytokines IL1β, IL6, IL10 and TNF-α in placental tissue, umbilical cord plasma and maternal plasma of the first and last trimester of gestation in obese (GO) and eutrophic (GE), to verify the associations between the contents of these cytokines and the placental protein and RNAm expression of FABP1, FABP3, FAT / CD36, FATP1, FATP2 and FATP4, as well as to test their correlation with characteristics of newborns. The recruitment of voluntary pregnant women occurred in two public institutions in Rio de Janeiro. In the first and third trimesters, samples of maternal blood were collected and, at delivery, umbilical cord blood and placental tissue. The biochemical analyzes consisted of Luminex, qPCR and Western blotting assays. The cytokine levels of maternal plasma in the first trimester correlated positively with the protein expression of several FA transporters. The protein content of FABP1 in placental tissue was lower in GO than in GE. The placental concentrations of FABP1 and FABP3 correlated positively with maternal weight gain and placental efficiency. The expression of FATP4 mRNA in placental tissue correlated positively with the head circumference. These results reinforce the importance of nutritional monitoring, prior to conception, to adjust maternal body mass index and the quality of lipid reserves, and, during the gestational period, to ensure optimal weight gain.
Keywords: placenta, fatty acids, transporters, cytokines, obesity, pregnancy

Read full


Title: Association Between Polymorphisms in Genes Related to Obesity and to Body Weight Changes and Food Consumption of Pregnant
Author: MAISA CRUZ MARTINS
Conclusion Paper: TESE
Abbreviation: MARTINS, M. C.
Date of Defense: 14/07/2017
Abstract: A large proportion of women are initiating gestation above the recommended weight, gaining excessive weight during pregnancy and retaining a high percentage of the gestational weight gain as a result of the global epidemic of obesity. Among the many factors contributing to overweight, genetic differences play an important role in the process of expressiveness of the obesity phenotype, probably through control mechanisms of satiety/appetite and food preferences. Objective: To study the association between single nucleotide polymorphisms (SNPs) in obesity-related genes (FTO - fat mass and obesity, rs9939609; MC4R - melanocortin 4 receptor, rs17782313; LEP – leptin, rs7799039 e LEPR - leptin receptor, rs1137101) and changes in body weight and dietary intake during gestation. Method: Prospective cohort of pregnant women attending a Municipal Health Center, located in the district of Tijuca in the city of Rio de Janeiro, in the following periods: 5-13, 20-26 and 30-36 gestational weeks and 30-45 days postpartum. Anthropometric measurements (weight and height) and plasma concentration of leptin were performed. Food intake data were obtained through a food frequency questionnaire (FFQ), referring to the pre-gestational and gestational periods. SNPs were analyzed by real-time polymerase chain reaction (PCR). The associations between the gene polymorphisms and the dependent variables (pre-gestational weight, gestational weight gain, postpartum weight retention, plasma leptin concentration and dietary intake) were investigated by longitudinal mixed effects regression models, multiple linear regression and Poisson regression models, adjusted for obstetric, socioeconomic, demographic factors and energy intake. Results: The FTO-rs9939609 (A-allele) gene SNP was significantly associated with pre-pregnancy overweight and with the highest percentage of energy from carbohydrates in pre-pregnancy. This SNP was also associated with the highest percentage of total energy and with the highest percentage of energy from ultraprocessed foods during pregnancy. The SNP of the MC4R-rs17782313 gene (C-allele) was positively associated with the highest percentage of energy derived from ultraprocessed foods throughout pregnancy. SNPs in the genes LEP-rs7799039 and LEPR-rs1137101 were not statistically associated with plasma concentrations of leptin throughout pregnancy and with food consumption components, however, the LEP-rs7799039 gene was significantly associated with the increased risk of excessive gestational weight gain. Conclusion: SNPs in xigenes related to obesity are associated with changes in body weight and dietary intake of pregnant women.
Keywords: Pregnant women; Single nucleotide polymorphisms; Pre-gestational weight; Gestational weight gain; Postpartum weight retention; Food consumption; Cohort study.

Read full


Title: Sistemic Blood Pressure and Serum Lipids During Pregnancy: Effect of Leptin and Leptin Receptor Genetic Polymorphisms and Association with Neonatal Outcomes
Author: DAYANA RODRIGUES FARIAS
Conclusion Paper: TESE
Abbreviation: FARIAS, D.R.
Date of Defense: 06/07/2017
Abstract: Studies have suggested that maternal blood pressure (BP) and serum lipids seem to be associated with birth weight (BW). However, little is known about how dietary intake, body mass index (BMI) and genetic polymorphisms interacts in the regulation of BP and lipid concentrations during pregnancy and in which way maternal lipid changes during pregnancy predicts BW. Objective: To evaluate the association between leptin (G2548A) and leptin receptor (Q223R e K109R) genetic polymorphisms with BP changes during pregnancy and post-partum, to investigate if dietary intake is a mediator in the association between these polymorphisms and serum lipid concentrations throughout pregnancy, and to test if lipids changes throughout pregnancy are associated with BW. Methods: Prospective cohort of 154 pregnant women conducted at a Public Health Center, in Rio de Janeiro, Brazil, that followed women during the three pregnancy trimesters and between 30-45 days postpartum. Serum concentrations (mg/dL) of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-c) and triglycerides (TG) were measured by enzymatic colorimetric method and low-density lipoprotein cholesterol (LDL-c) was calculated. The systolic (SBP) and diastolic (DBP) blood pressure (mmHg) was measured using an automatic sphygmomanometer. The leptin (G2548A) and leptin receptor polymorphisms (Q223R e K109R) were genotyped using real time polymerase chain reaction (PCR) method. BW was obtained from the child vaccination booklet and BW z-score for gestational age and sex was estimated according to the Intergrowth-21st standard. Statistical analyses included adjusted linear mixed-effect and linear regression models, reporting β coefficients and 95% confidence interval (CI). Results: The adjusted models showed that: (1) Women carrying the G-allele of G2548A polymorphism presented higher BP levels compared to the AA genotype (βSBP=4.5; 95%CI=1.0;8,0; βDBP=2.9; 95%CI=0.1;5.8). There was a significant interaction between leptin polymorphism and BMI, in which the effect of BMI on increasing BP was steeper in women homozygous for the A-allele, compared with those who had at least one G-allele. We did not find significant associations between polymorphisms Q223R and K109R and BP, in the adjusted models (2) Women with the AA genotype presented higher concentrations of LDL-c (β=16.0; 95% IC=-0.2;32.2) at the second and third trimester, a higher increase/gestational week in TG (β=4.5 vs. β=8.4) and TC (β=6.2 vs. β=7.8) and higher means of total energy and lipid intake, compared with those with at least on G-allele. We did not find significant associations between polymorphisms Q223R and K109R and lipids concentrations during pregnancy, in the adjusted models. (3) The pregnancy variation of maternal LDL-c concentrations (β=0.36; 95% CI=0.16;0.56) was positively associated with BW z-score, while HDL-c concentrations (β=-2.0; 95% CI=-3.1;-0.8) were negatively associated. Conclusions: The leptin polymorphism (G2548A) was associated with the variation of BP levels and presented a significant interaction with maternal BMI. Women with AA genotype of G2548A polymorphism presented higher rate of change/gestational weeks of TG and TC, higher concentrations of LDL-c at the second and third trimesters and higher means of total energy and lipid intake, but the association between the polymorphism and serum lipids was not explained by the dietary intake. The variation of maternal LDL-c and HDL-c concentrations was significantly associated with BW. We did not observed significant associations between Q223R and K109R polymorphisms with BP or lipids changes during pregnancy in the adjusted models.
Keywords: Prospective cohort; Lipids; Leptin, Blood pressure; Pregnancy, Birth weight; Genetic polymorphisms.

Read full


Title: Metabolic Syndrome and its Risk Factors in Schools Aged 6 to 10 Years of Age Macaé-RJ
Author: FABIANA DA COSTA TEIXEIRA
Conclusion Paper: TESE
Abbreviation: TEIXEIRA, F. C.
Date of Defense: 19/05/2017
Abstract: Metabolic syndrome (MetS) and its risk factors (RF) may begin at early ages and are correlated with structural, functional and metabolic changes in childhood and with non-communicable chronic diseases in adulthood, including cardiovascular diseases and type 2 diabetes melittus. Although studies to evaluate SM and its FR in juvenile group are growing, they are not so frequent in children, especially in developing countries. Objectives: The aim of the present study was to identify the prevalence of RF to the diagnosis of MetS and its association with nutritional status of schoolchildren from 6 to 10 years old. Methods: A cross-sectional study was conducted in 505 students aged between 6 and 9 years, 11 months and 29 days, of both sexes, in nine schools in the city of Macaé, Rio de Janeiro. The study was conducted from March 2013 to November 2014 by trained staff of the laboratory of Nutritional Epidemiology, UFRJ, Macae. Data collected: bodyweight (Kg), height (m) and waist circunference (cm), blood pressure (mmHg), triglycerides, HDL-cholesterol (mg/dl) and fasting blood glucose (mg/dl). The MetS was defined according to the Third Report of the National Cholesterol Education Program, with the cut-off points adapted for children. Results: The sample included 505 schoolchildren 221 (43.8%) girls and 284 (56.2%). By nutritional status, 14 (2.8%), 312 (61.8%), 85 (16.8%) and 94 (18.6%) had underweight, normal weight, overweight and obesity, respectively. At least one RF was present in 61% (n=308) of the sample. Hypertriglyceridemia was the most prevalent RF, observed in 29.3% (n=148) of the sample. The low HDL-c, more prevalent in males than in females, was the only RF that showed a significant difference between sexes (p = 0.03), and was not associated with nutritional status (p =0.43). There was higher prevalence of RF in overweight/obese schoolchildren compared to those with normal weight. The prevalence of one, two and three RF (MetS) were 34.7% (n = 175), 21.0% (n = 106) and 5.3% (n = 27), respectively. Two RF were more present in overweight (28.2% 95%CI 19.0; 39.0) and obese (41.5% 95%CI 31.4; 52.1) compared to normal weight children (13.5% 95%CI 9.9; 17.8). Three or more RF were more frequent among obese (25.5% 95%CI 17.0; 35.5) in relation to overweight (2.4% 95%CI 0.2; 8.2) and normal weight children (0.3% 95%CI 0; 1.7). Conclusion: This study indicated a high prevalence of RF that contributes to MetS and its association with the magnitude of the weight excess. Therefore, it is indicated to identify children with cardiometabolic RF regardless of MetS’ diagnosis. The data reinforce the importance of prevention and treatment of excess weight in order to minimize the risk of MetS and related diseases in adulthood.
Keywords: children, cardiovascular risk, metabolic syndrome, abdominal obesity, dyslipidemia, hypertension

Read full


Title: Serum Concentrations of Retinol, Beta-Carotene, and Zinc in a Cohort of Women with Second Stage Breast Cancer and Treatment Toxicity
Author: CINTIA LETICIA DA SILVA ROSA
Conclusion Paper: TESE
Abbreviation: ROSA, C. L. S.
Date of Defense: 29/03/2017
Abstract: The procedures used to treat breast cancer are able to raise free radical production, increase oxidative stress and deplete antioxidants. Objective: To analyze the serum concentrations and synergism between retinol, β-carotene and zinc in women with breast cancer, according to the stage of the disease and the acute and late toxicity of the radiotherapy, considering different modalities of treatments prior to radiotherapy. Methods: This is a longitudinal study in a cohort of women with breast cancer evaluated at the pre-treatment periods, 7 and 120 days after radiotherapy. Patients were divided into 3 groups: G1 undergoing conservative surgery, G2 undergoing chemotherapy, and G3 undergoing conservative surgery and chemotherapy. The serum concentrations of retinol and β-carotene were quantified by high performance liquid chromatography with ultraviolet detector, and zinc by atomic absorption spectrophotometry. Staging of breast cancer was based on the classification of malignant tumors TNM, proposed by the International Union Against Cancer (UICC). Radiotherapy treatment was performed using teletherapy using linear accelerator with 6Mv energy with total dose ranging from 50-50.4 Grays (Gy). Acute toxicity was assessed according to the acute toxicity scale of the Radiation Therapy Oncology Group (RTOG) and late toxicity RTOG / EORTC (European Organization for Research and Treatment of Cancer). The toxicities were divided into groups I and II, composed of groups of toxicities grades 0 and 1 and 2 and 3, respectively. Results: A total of 230 patients were evaluated, with a mean age of 63.6 years (SD + 9.3). In the sample, stage II of breast cancer predominated. It was observed that serum concentrations of the micronutrients evaluated were reduced as the level of disease staging increased. At all stages and in all groups, the highest percentages of deficiency occurred in serum concentrations of zinc and retinol. Surgery alone had a greater negative impact on serum retinol concentrations. Surgery combined with chemotherapy had a greater negative effect on the serum concentrations of β-carotene and zinc. Considering the groups studied at T0, the highest percentage of retinol deficiency was in G3 with 21.3%, betacarotene in G1 38.5% and zinc also in G1 corresponding to 30.8%. A statistically significant reduction was observed in the mean of all the micronutrients studied, when T0 was compared one week after the radiotherapy treatment (T1). After 120 days all the nutrients evaluated increased significantly when T1 and T2 were compared. Acute toxicity was observed in 67.2% of the cases and the late toxicity of 100% of the sample, both belonging to group I. Considering the acute toxicity, there were higher serum concentrations of retinol, beta-carotene and zinc in group I when compared to Group II. With regard to late toxicity, there was a significant difference in the majority of treatment groups prior to radiotherapy, with the exception of zinc. Conclusion: Among the treatments prior to radiotherapy, surgery combined or not with chemotherapy exerted a greater negative impact on all micronutrients studied. There was a significant reduction of all micronutrients in T1, regardless of the type of previous treatment. In acute and late toxicity, there was a reduction of antioxidants in the groups at the highest grade, demonstrating the need for monitoring and maintenance of the nutritional status of these antioxidants throughout cancer treatment.
Keywords: Breast cancer, staging, surgery, chemotherapy, radiation therapy, toxicity, vitamin A, zinc.

Read full


Title: Membrane Microdomains and Lipid Profile of Human Breast Cancer Cells MCF-7 and MDA-MB-231 Under the Action of Resveratrol
Author: LUCIANA DOS SANTOS GOMES
Conclusion Paper: TESE
Abbreviation: GOMES, L. S.
Date of Defense: 10/03/2017
Abstract: Cancer is the second largest cause of death worldwide. For the biennium 2016-2017 the estimate for Brazil indicates the occurrence of approximately 600 thousand new cases, with female breast cancer representing 28.1%. Resveratrol (Resv) is a phytochemical which can act at the three stages of carcinogenesis through several molecular targets. Frequently its effect is related to its uptake by membrane microdomains. These regions are composed of cholesterol, sphingolipids and proteins, such as flotillins (flot), which are associated with cancer’s poor prognosis when overexpressed. Thus, the aim of this study was to investigate the effect of Resv on the membrane microdomains and the lipid profile of human breast cancer cell lines MCF-7 and MDA-MB-231. The cytotoxicity was checked by MTT (methylthiazolyldiphenyl-tetrazolium bromide) assay, after 24 hours of treatment. Resv effect on flot-1 was analyzed by dot-blotting after 24 hours of treatment. Flot-1 and flot-2 expression was measured by Western Blotting analysis, and the regulation of the gene expression was analyzed by quantitative reverse transcriptase polymerase chain reaction (RT-qPCR). Immunofluorescence microscopy was used to screen for flot-2 in MDA-MB-231 cell. Study of the lipid profile was performed by high performance thin-layer chromatography (HPTLC) and thin-layer chromatography (TLC). Fatty acids (FA) identification occurred by gas chromatography with mass spectrometry (GC/MS). Phospholipid (PL) biosynthesis was performed using radioactive inorganic phosphate and TLC. Lipid metabolism were analyzed by Western Blotting, checking diacylglycerol O-acyltransferase 2 (DGAT2), fatty acid synthase (FAS), acetyl-CoA carboxylase β phosphorylated (ρACCβ), 5' adenosine monophosphate-activated protein kinase (AMPKα) and phosphorylated (ρAMPKα) enzymes. Results demonstrate that for MCF-7 there was cytotoxic effect of Resv from 50 μM and for MDA-MB-231 from 100 μM. Concentrations of 80 and 200 μM were used in subsequent assays for MCF-7 and MDA-MB-231, respectively. The data obtained by Western Blotting did not indicate significant differences for flot-1 and flot2, however, in immunofluorescence microscopy it was observed a 13.4-fold increase in the area corresponding to flot-2. RT-qPCR indicated a sharp increase in the gene expression of flot-1 and flot-2 in MDA-MB-231. Significant reduction (p < 0.05) of cholesterol ester and an increase of FA (2.9-fold) was observed im MCF-7. In MDAMB-231 FA increased 5.3-fold. The predominant saturated FA in both cells were palmitic acid and stearic acid and unsaturated FA were oleic acid and elaidic acid. MCF-7 cells showed an increase of 65% of phosphatidylethanolamine. Sphingomyelin and lysophosphatidylcholine (LPC) showed the highest reduction percentage, 57% and 40%, respectively. MDA-MB-231 showed reduction of phosphatidylcholine (63%) and LPC (52%). Resv reduced ρACCβ (3.3-fold, p < 0.05) and AMPKα / ρAMPKα (1.5fold, p <0.04) in MCF-7 and in MDA-MB- 231 at 111.8 (p < 0.03) and 1.2 (p < 0.005) times, for ρACCβ and AMPKα / ρAMPKα, respectively. The results obtained allow us to verify that Resv increases the expression of flot and FA in MCF-7 and MDA-MB231, and that the cytotoxic effect of the phytochemical remains. This indicates that although the compound is favoring events related to tumor progression, other molecular targets of Resv might be active, contributing to its anticancer action. With this, new studies are needed to analyze by what mechanisms Resv acts under these conditions. Keywords: Resveratrol, flotillin, detergent-resistant membranes, breast cancer, lipid metabolism

Read full


Sample Description
Sample Description