PPGB - Mestrado em Bioquímica
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Master Thesis Beta-Glucanas extraídas do fungo Caripia montagnei (Berk.) Kuntze: uma análise das suas propriedades antioxidante, antitumoral, antiangiogênica e anti-inflamatória de forma isolada e em sinergia com AINEs, L-NAME e ligantes de PPAR alfa e gama(Universidade Federal do Rio Grande do Norte, 2011) Castro, Allisson Jhonatan Gomes; Leite, Edda Lisboa; Cruz, Ana Katarina Menezes da; http://lattes.cnpq.br/9810605697247961; Martins, Daniella Regina Arantes; http://lattes.cnpq.br/3695151190501921; http://lattes.cnpq.br/7712988818433906; http://lattes.cnpq.br/6781850620408652Glucans are homopolymers of D-glucose with bioactive properties with direct influence o activation of leukocytes and antitumoral, antioxidant and immunomodulatory activities. The aim of, this paper was to verify the efect of p-glucans extracted from the mushroom Canpia m 9 the inhibition of the inflammation, angiogenesis and free radicais. The fraction polysaccna showed carbohydrate (98.5%), protein (1.5%) and 0.1 mg / mg of phenolic compounds. Its monosaccharide composition showed predominantly glucose by means of HPLC, with nuclear magnetic resonance (nmr) confirmed the presence of D-glucose to 103.5 ppm in P configuratio. The viability of fibroblasts in the presence of glucans from C. Montagnei at 25 uq was 83.3±17,6%, with high cytotoxicity on tumoral cells (HeLa) at 100 ug (94.36±2.3%). The anti-edematous activity observed at 90 mg/kg of Carípia montagnei was 90.7 ± 3.5%. The synergy with parecoxib showed was 81 3+7.4%, 100±12% to L-NAME and 97.8+10% with PFOA. In study made with carrageenan 91.043±7,3%. The production of nitric oxide with Caripia polysacchandes was (48.2%) was better than the activity of indomethacin. The glucans of C. montagnei showed significant antiangioge activity with 1 mg/egg (p>0.05). In total antioxidant activity and reducmg power was foundI for the relative antioxidant activity of BHT 57% and 84%, respectively. The evaluation of the inhibition of hydroxyl radical (1 mg/ml) was 38±0.01%, while for the lipid peroxidation was observed an inhibition of 37 6±0 16%. The present results showed that p-glucans from Carípia montagnei have inhibitory activity in these models of inflammation, angiogenesis and inhibition of free radicais.Master Thesis Comparação do potencial antioxidante in vitro, em cultivo de células e in vivo de fucoidans obtidos comercialmente(Universidade Federal do Rio Grande do Norte, 2021-10-08) Silva, Maylla Maria Correia Leite; Câmara, Rafael Barros Gomes da; Rocha, Hugo Alexandre de Oliveira; 0000-0003-2252-1221; http://lattes.cnpq.br/4651814546820796; http://lattes.cnpq.br/9200036511632745; http://lattes.cnpq.br/3703561529618882; Lucena, Eudes Euler de Souza; http://lattes.cnpq.br/6969519171915250; Pinhal, Maria Aparecida da Silva; http://lattes.cnpq.br/7511274763693292Fucoidans are sulfated L-fucose-rich polysaccharides that are extracted from brown seaweed. These polymers are already commercially available and have been evaluated for various purposes. One of the most evaluated activities is the antioxidant, among the fucoidans that have already been analyzed for antioxidant capacity are the fucoidans from brown seaweed: Undaria pinnatifida; Macrocystis pyrifera and Fucus vesiculosus. However, the antioxidant activity of fucoidans from the respective seaweeds have not yet been compared. Thus, the objective of this work was to identify which of these fucoidans, which are commercially obtained (Sigma-Aldrich), has the best antioxidant activity. Therefore, the fucoidans were analyzed for their physicochemical composition, the results obtained showed that they are rich in fucose and sulfate. And regarding the apparent molecular mass, it was found that it ranged from 70.4 to 168.5 kDa. The fucoidan from U. pinnatifida being the largest polymer. In vitro antioxidant tests all showed fucoidans with moderate activity, except for the H2O2 scavenging assay, where all fucoidans exhibited high activity. When fucoidans were evaluated under macrophage cell culture conditions (RAW 264.7) by MTT (3-(4,5-dimethylthiazol-2-yl) 2,5- diphenyl tetrazolium bromide) and live/dead reduction assays, after induction of oxidative stress with H2O2, it was observed that fucoidans reduced the effect of H2O2 on the metabolism of these cells, especially the fucoidan from F. vesiculosus (5 mM). The data obtained in vitro and with cells provided support for the performance in vivo tests with the Danio rerio (zebrafish) embryo model. Two analyzes were carried out: determination of the survival rate of the embryos and quantification of the fluorescence intensity of the hatched larvae. It was observed that in the presence of fucoidans and H2O2, the survival rate of the embryos was little affected, compared to the treatment that received only H2O2 (0.4 mM), where there was a 50% reduction in the survival rate. The evaluation of larvae treated with fucoidan and with H2O2 (0.4 mM) showed that the lowest percentage of fluorescence intensity was observed with those treated with F. vesiculosus and M. pyrifera fucoidan. Considering the results of the tests carried out, fucoidan from M. pyrifera is pointed out as the fucoidan with the best antioxidant potential, especially in vivo tests. Thus, fucoidan from M. pyrifera is the most promising in terms of antioxidant activities.Master Thesis Caracterização de comunidades e isolados bacterianos obtidos a partir de amostra de petróleo de pré-sal brasileiro sob cultivo em presença de CO2(Universidade Federal do Rio Grande do Norte, 2021-07-20) Barbalho, Kamilla Karla da Silva; Lima, Lucymara Fassarella Agnez; http://lattes.cnpq.br/1083882171718362; http://lattes.cnpq.br/8583882001958993; Thompson, Fabiano Lopes; http://lattes.cnpq.br/4148145822928884; Melo, Vânia Maria Maciel; http://lattes.cnpq.br/1572504650930605Microorganisms represent the largest biomass on Earth and are directly related to biogeochemical cycles. It has been described that microbial activity affects the production chain related to the oil and gas sector, being of interest the knowledge about the microbiology associated with reservoirs, aiming the optimization of the production chain, as well as the mitigation of environmental impacts due to environmental contamination by hydrocarbons. The Brazilian pre-salt reservoirs are extreme environments with high carbon dioxide (CO2) concentration and high depth, whose microbiology is still poorly understood. The objective of this work was to characterize bacterial communities and isolates obtained from a sample of pre-salt oil submitted to cultivation at high CO2 concentration. From an oil sample, six consortia were obtained and analyzed, organized in two groups: three undefined microbial consortia grown at high CO2 concentration and the other three obtained from the same sample under atmospheric aeration and later submitted to high CO2 concentrations. The microbial consortia were characterized for: i) biosurfactant (BS) production; ii) growth curve in the presence of hydrocarbons as carbon source; and iii) taxonomic and functional characterization by metagenomic sequencing. Functional assays of the consortia showed that three are able to produce BS molecules capable of generating emulsion, whereas none of the consortia is able to disperse oil. The growth curves showed that five consortia are able to grow in the absence of external carbon sources, indicating the occurrence of inorganic carbon fixation, which was corroborated by the metagenomic sequencing of the consortia via identification of sequences assigned to the genus Ochrobactrum, which is able to fix carbon. In the consortia, the most abundant phyla were Proteobacteria, Actinobacteria, and Firmicutes, and the consortia submitted to high CO2 concentration, after growing in O2, were more diverse than the consortia grown in high CO2 concentration. As for the metabolic profile, the most enriched pathways in the consortia grown in CO2 were the amino acid metabolism and carbohydrate metabolism pathways. Nineteen bacterial isolates were obtained from the consortia, and nine were characterized and tested. For the isolates the following assays were performed: i) BS production, ii) hydrocarbon degradation capacity, iii) cellular hydrophobicity, and iv) enzymatic activity of hydrolases. Of these, five were able to degrade between 50% and 80% of the xylene and toluene fractions, and eight others degraded 100% of the gasoline, diesel and kerosene fractions. It is concluded that the growth of undefined consortia in O2 leads to a greater diversity of microorganisms, that CO2 inhibits BS production, and that isolates and microbial consortia from the pre-salt show promise as a source of bioremediation products, as well as hydrolases with potential industrial application.Master Thesis Atividade antibacteriana de magnetita (Fe3O4) isolada e associada a campos eletromagnéticos sobre Escherichia coli DH5a pVAX-1 (resistente à canamicina)(Universidade Federal do Rio Grande do Norte, 2021-06-30) Santos, Sunamita da Silva Barbosa; Silva, Marcelo de Sousa da; http://lattes.cnpq.br/1295430560645312; http://lattes.cnpq.br/6988181736543670; Melo, Maria Celeste Nunes de; http://lattes.cnpq.br/0580551464788795; Barbosa, Júlio César Pereira; http://lattes.cnpq.br/0437190557200470Due to the increase in bacterial resistance in recent years, research and development of alternative antibacterial therapies has become necessary and, in this sense, the use of magnetic fields has been explored as it is a non-pharmacological therapy and, therefore, probably exempt of the molecular effects of the resistance phenomenon. At the same time, nanoparticles with magnetic properties, especially magnetite (Fe3O4), are still little explored in terms of their antibacterial potential. In this context, the present work evaluated the intrinsic antibacterial properties of magnetite nanoparticles, synthesized by high-energy milling, and their association with a variable external magnetic field of 70 mT at 10 Hz on Escherichia coli DH5α pVAX-1. After synthesis, the nanoparticles were characterized as to their composition and primary size, by X-ray diffraction (XRD); as to its hydrodynamic size, by dynamic light scattering (DLS) and as to its surface charge, by zeta potential. Subsequently, in vitro assays were performed, in which alterations in the bacterial inhibition curve were investigated; in bacterial morphology, using microscopy (SEM), and in bacterial surface load, using zeta potential. The association between the antibiotic kanamycin and magnetite was also evaluated regarding the potentiated effect between the substances. Exposure to isolated magnetite, or combined to a magnetic field, generated significant bacteriostatic effects, in addition to morphological changes in the bacterial surface and an increase in the zeta potential of the suspension. The use of magnetite nanoparticles also potentiated the inhibitory effect of the antibiotic kanamycin, pointing to a second option for use as an adjunct to antibiotic therapy. The results presented reveal important antimicrobial potential and additional in vivo tests are needed to prove the importance of the effects observed in a biological system.Master Thesis Efeito do inibidor de tripsina isolado da semente de tamarindo (Tamarindus indica L.) nanoencapsulado sobre parâmetros bioquímicos e histopatológicos do fígado de ratos Wistar alimentados com dieta de alto índice glicêmico(Universidade Federal do Rio Grande do Norte, 2021-05-28) Aguiar, Ana Júlia Felipe Camelo; Morais, Ana Heloneida de Araújo; Passos, Thais Souza; 0000-0003-2054-1544; http://lattes.cnpq.br/9685790797554876; http://lattes.cnpq.br/1233944493334651; http://lattes.cnpq.br/3937321074677193; Moura, Sérgio Adriane Bezerra de; http://lattes.cnpq.br/1323062374722389; Ferreira, Ederlan de Souza; http://lattes.cnpq.br/4952207117831158There has been a nutritional transition based on diets composed of foods with a high glycemic index and a high glycemic load and ultra-processed foods worldwide. This, associated with other factors, is crucial for increasing the obesity prevalence and various diseases, including the disease Non-Alcoholic Fatty Liver (NAFLD). NAFLD ranges from steatosis and steatohepatitis to severe conditions, such as cirrhosis and hepatocellular carcinoma. NAFLD treatment is a clinical necessity not answered. Thus, several studies have sought new therapies not only for NAFLD but also for obesity. Among them, we highlight the findings related to the trypsin inhibitor isolated from tamarind seeds (TTI) nanoencapsulated in chitosan and whey protein isolated (ECW). Therefore, the present study aimed to investigate the effect of ECW on the liver status of Wistar rats fed a high glycemic index and high glycemic load diet (HGLI diet). First, nanoparticles with and without TTI, respectively ECW and CW, were obtained by the technique of nanoprecipitation in an organic solvent and, characterized by morphology (SEM), particle size (Laser diffraction), chemical interactions (FTIR), and efficiency of encapsulation. The animals were divided into four groups, containing five animals, each treated for ten days with 1. HGLI diet and water (1 mL), 2. standard diet and water (1 mL), 3. HGLI diet and ECW (12.5 mg/kg in 1 mL), and 4. HGLI diet and CW (10.0 mg/kg in 1 mL). The hematological biochemical parameters of renal and hepatic involvement were evaluated (liver enzymes, AST/ALT ratio), lipid profile, fasting glucose, insulin and insulin resistance, and β-cell homeostasis model (HOMA-IR and HOMA-β, respectively). Besides, liver scores FIB-4 (Fibrosis-4 Index for Liver Fibrosis) and APRI (AST to Platelet Ratio Index) were also investigated, and the liver histopathology of the animals was evaluated. Given the results, smooth, spherical nanoparticles were obtained in nanometric diameter (120.7 nm) containing TTI. In animals, ECW reduced (p < 0,05) fasting blood glucose levels by 17%, AST in 39%, and alkaline phosphatase in 24%, also significantly reduced (p < 0,05) the two scores of assessment of liver fibrosis, APRI, and FIB-4, and presented a better aspect of liver morphology when compared to the other groups. Thus, it was possible to show that the ECW promoted a new functionality for the isolated TTI, promoting benefits on the liver injury caused by the HGLI diet, considering the biochemical, morphological, and liver fibrosis evaluation parameters.Master Thesis Avaliação de extratos ricos em polissacarídeos, obtidos a partir de cebolas amarelas e roxas (Allium cepa, Linn), quanto ao seu potencial antioxidante e modulatório da cristalização de oxalato de cálcio (CaOx)(Universidade Federal do Rio Grande do Norte, 2021-01-29) Machado, Raynara Iusk de Araújo; Rocha, Hugo Alexandre de Oliveira; ; http://lattes.cnpq.br/4651814546820796; ; http://lattes.cnpq.br/3979838028433164; Câmara, Rafael Barros Gomes da; ; http://lattes.cnpq.br/9200036511632745; Lima, Jailma Almeida de; ; http://lattes.cnpq.br/9077722100091492The onion (Allium cepa Linn.), plant of the Amaryllis family (Amaryllidaceae), is one of the most important vegetable species in the world, standing out for being the second with greater added value. Its structure is mostly made up of carbohydrates, whose pharmacological applications have been studied and verified for a long time. Thus, the objective of this work was to obtain extracts rich in polysaccharides, from yellow (YAC) and red onions (PAC), characterize them chemically - through the quantification of sugar, protein, sulfate, and phenolic compounds - and structurally - by procedures such as HPLC and infrared chromatography - in search of compounds that can provide greater added value and applicability to these plants. Following this search, tests were carried out to evaluate the extracts as to the antioxidant potential and potential modulator of the formation of calcium oxalate crystals (CaOx). It was observed that the extracts obtained are mainly composed of heteropolysaccharides, which have the following monosaccharides in their structures: glucose, galactose and uronic acids (PAC) and glucose, galactose, uronic acids and arabinose (YAC). In addition to the carbohydrate portion, the presence of sulfates and low levels of proteins and phenolic compounds were also identified in the extracts. However, despite the considerable amount of sulfate found in the samples, there is no evidence that onions synthesize sulfated polysaccharides. To evaluate the antioxidant potential of YAC and PAC extracts, five tests were performed: total antioxidant capacity (CAT), reducing power, copper chelation, iron chelation and hydroxyl radical scavenging. In three of the five tests performed, better results were obtained for the YAC extract. These best values were identified specifically in the tests: CAT (110.8 mg of AA per gram of the extract), reducing power (48% of activity) and iron chelation (80.01% of chelating activity). Regarding the modulatory activity of CaOx, positive results were found for the YAC extract, which was effective in directing crystallization to the formation of small-sized dihydrate calcium oxalate crystals (COD), which could, in biological systems , facilitate the process of excretion of the crystals in the urine and decrease the ability of the crystals to bind to the epithelial cells of the renal tubules causing damage. The results found in this work are promising and open the way for further studies that can evaluate and confirm such potentials in vivo.Master Thesis Avaliação do potencial antioxidante de Condroitim sulfato extraído das vísceras de tilápia Oreochromis niloticus(Universidade Federal do Rio Grande do Norte, 2020-08-27) Medeiros, Luiz Henrique Costa de; Chavante, Suely Ferreira; Souza, Giulianna Paiva Viana de Andrade; ; http://lattes.cnpq.br/9747224552107070; ; http://lattes.cnpq.br/3440814329803472; ; http://lattes.cnpq.br/9786859259093914; Bezerra, Iglesias de Lacerda; ; http://lattes.cnpq.br/9190547635625352; Filgueira, Luciana Guimaraes Alves; ; http://lattes.cnpq.br/9951316929526841Glycosaminoglycans are polysaccharides present in the cell surface and extracellular matrix of the vast majority of animals. They perform several biological functions, such as cell growth, differentiation, antioxidant, neuroprotec and others. Among glycosaminoglycans, chondroitin sulfate (CS) stands out for its antioxidant effect and can also be found in many fish species, such as tilapia (Oreochromis niloticus) whose cultivation represents an important economic activity in Natal (RN / Brazil). ), eliminating a big quantity of viscera as a waste of its production, daily. Thus, the following study uses the tilapia wastes that have been obteined in free markets in city of Natal, as raw material for the extraction and purification of chondroitin sulfate, in addition to evaluete its antioxidant effect. For this, the residues were exposed to an enzymatic proteolyse process, followed by complexation and decomplexing with Lewatit resin, obtaining a pool of glycosaminoglycans that have been used in purification procedures like acetone fractionation and ion exchange chromatography, accompained by agarose gel electrophoresis and enzimatic degradation. Subsequently, antioxidant tests were performed to quantify the ability of CS to inhibit reactive species intracellular and in vitro, since the imbalance in their quantities has a role in clinical conditions such as autoimmune, neurodegenerative diseases and cancer. For this, methods such as inhibition of intracelular ROS (DCFH-DA), chelation of ferrous and cupric ions, inhibition of superoxide and hydroxyl radicals, in addition to inhibition of nitric oxide and lipid peroxidation were used. As a result, it was possible to observe that in each kilogram of tilapia viscera dry mass it is possible to extract approximately 69mg of purified chondroitin sulfate, being 59% composed of chondroitin-4-sulfate. By observing the values obtained in the antioxidant tests, it was possible to highlight an inhibition of up to 52% of the reactive intracellular species in culture of RAW 264.7 cells, without demonstration of cytotoxicity up to the concentration of 200mg /mL of chondroitin. The other tests showed that this inhibition occurs due to several factors, ranging from the inhibition of metals and free radicals (p<0,05), to the probable participation in secondary pathways, such as the activation of antioxidant proteins. The results obtained in this study strengthen the extraction and use of glycosaminoglycans, especially chondroitin sulfate, helping to understand the biological effects of chondroitins from multiple sources, emphasizing their antioxidant effect and providing a disposal alternative for the biological waste produced by commercial tilapia production, opening perspectives for possible uses of this molecule in research and industry.Master Thesis Caracterização de frações ricas em polissacarídeos sulfatados da alga marrom Dictyopteris delicatula e avaliação do seu potencial antiurolítico, a partir da determinação de suas atividades antioxidante e moduladora da cristalização de oxalato de cálcio(Universidade Federal do Rio Grande do Norte, 2020-03-27) Batista, Lucas Alighieri Neves Costa; Rocha, Hugo Alexandre de Oliveira; ; ; Scortecci, Kátia Castanho; ; Silveira, Raniere Fagundes de Melo;Fucans and fucoidans are polysaccharides that contain sulfated L-fucose in their constitution and are found in brown algae, several pharmacological activities are attributed to these molecules such as: anticoagulant, antithrombotic, antiproliferative, anti-nociceptive, osteogenic and others. Wide presence of carboxyl and sulfate groups in fucans and fucoidans gives these molecules a high negative charge, which is a common characteristic of inhibitors of calcium oxalate (CaOx) kidney stones formation, so fucans and fucoidans are probably antiurolithic agents, which has already been demonstrated in some studies. Antiurolithic activity is also related to reduction of oxidative stress in renal tubules therefore fucans and fucoidans are also considered antiurolytic due to their antioxidant action. Considering the negative charge and antioxidant activity as a key factor for antiurolithic activity, the present study aims to evaluate the antiurolithic potential of polysaccharides extracted from the macroalgae Dictyopteris delicatula - a macroalgae widely found throughout the northeastern coast - through analysis of these polysaccharides effect in CaOx crystallization and their potential to prevent oxidative stress in MDCK cells. D. delicatula total polysaccharides were extracted by proteolysis and fractionated by precipitation with increasing volumes of acetone in five different fractions: F0.5v, F0.7v, F1.0v, F1.5v and F2.0v. Fractions compositions was determined by physical-chemical analysis, which proved they were sulfated polysaccharides rich fractions (SPRF). CaOx crystallization was induced in vitro by adding sodium oxalate to a solution of calcium chloride, which contained or not one of the FRPS solution. When formed in the presence of FRPS, CaOx crystals are smaller, more numerous and with a highly negative surface charge, with emphasis on the F0.5v modulatory activity, whose presence in crystallization increased 1721 times the number of crystals formed, reduced by more than 90 % crystals size, increased the negative charge on the surface from -7 mV to -35 mV and completely prevented the formation of monohydrated CaOx crystals, based on these results the F0.5v was chosen to be better evaluated. F0.5v was fractionated by ion exchange chromatography and five subfractions were obtained: 0.2 M, 0.4 M, 0.5 M, 0.6 M and 1.0 M. The chemical characterization of F0.5v and its subfractions revealed that F0.5v is consisting of alginate, heterofucans and laminarin. The antioxidant and CaOx crystallization modulating activities of none of the subfractions were superior to the activities of unfractionated F0.5v, indicating that F0.5v is more efficient when in the native form. Regarding the antioxidant activity, F0.5v was able to scavenge 79% of hydroxyl radicals, 10% of superoxide radicals, 22% of nitric oxide and have antioxidant capacity equivalent to 31 mg of ascorbic acid, in addition it has antioxidant activity ex vivo and increased up from 55% to 100% the viability of renal cells exposed to hydrogen peroxide, as for the modulation in crystallization, F0.5v, even in small concentrations like 0.01 mg / mL, promotes heterogeneous nucleation and the formation of small, dihydrated and unaggregable crystals. Considering the antioxidant and modulatory activity of CaOx crystallization of F0.5v we conclude this compound has the potential to prevent the formation of renal CaOx stones.Master Thesis Avaliação do efeito da inibição química das funções redox e reparo de APE1/Ref-1 sobre a transcrição gênica durante a inflamação(Universidade Federal do Rio Grande do Norte, 2020-04-14) Oliveira, Thais Teixeira; Lima, Lucymara Fassarella Agnez; Oliveira, Riva de Paula; Pinto, Nadja Cristhina de SouzaAPE1/Ref-1 is a protein that has two functional domains in mammals. The N-terminal domain is capable of reducing transcription factors, such as NF-kB, AP-1 and HIF-1 and increase its DNA binding capabilities and the expression of its target genes. The C-terminal domain, on the other hand, is responsible for the endonuclease activity of APE1/Ref-1, which is the main endonuclease of the base excision repair (BER) pathway that acts in the repair of apurine/apyrimidine (AP) sites. Due to this, APE1/Ref-1 is a potentially well-studied therapeutic target, especially in relation to cancer. Recently, it has been demonstrated that the presence of 8-oxoG, the main oxidized DNA damage, in the promoters of some genes can result in the induction of gene expression in the presence of DNA glycosylase OGG1 and APE1/Ref-1. Thus, APE1/Ref-1 also regulates gene expression in a manner dependent on its endonuclease function. However, the role of each of the APE1/Ref-1 functions on transcriptional regulation is not yet fully elucidated. In a previous work by our group, it was observed through RNAseq, that the inhibition of APE1/Ref-1 endonuclease activity by methoxyamine changes the expression of several genes during inflammation as well as decreases the expression of cytokines and chemokines in LPS-treated monocytes. Thus, the objective of this research was to verify which other genes have expression affected by the inhibition of AP repair, as well as to identify through bioinformatics tools possible regulators and partners of APE1/Ref-1 in this response. For this, RNA-seq was re-analyzed using bioinformatics tools and updated databases. In addition, the genes differentially expressed after treatment with methoxyamine were analyzed for their functions and possible regulators of their expression. The analysis of the transcriptome indicated that treatment with methoxyamine increases the expression of genes related to inhibition of the mTORC1 pathway and the protein polyubiquitination. On the other hand, it reduces the expression of genes related to cell cycle progression, transcription, expression of mitochondrial genes, biosynthesis of prostaglandins and Toll-like signaling independent of MyD88. In addition, these genes were compared with those differentially expressed after treatment with E3330 (specific inhibitor of redox function) under the same conditions. The main regulators of common genes as well as their targets were analyzed for their expression using q-PCR and Western blot. It was observed that both, the repair and redox function of APE1/Ref-1, are able to decrease the transcription of NRF1, GABPA and ELK1 target genes. These genes are related to mitochondrial biogenesis, rRNA processing and ribosomal biogenesis. In addition, it was observed that both APE1 / Ref-1 functions similarly decrease the expression of genes related to mitochondrial biogenesis, as well as being able to decrease Myc expression. On the other hand, the expression of ribosomal proteins was mainly affected by E3330, while inhibition of DNA repair was able to decrease the expression of the 47S pre-rRNA without affecting the processing of this RNA. Finally, we can conclude that both functions of APE1/Ref-1 may be involved in the regulation of genes related to mitochondrial biogenesis, but play different roles in the regulation of genes related to the processing and transcription of rRNA.Master Thesis Uma nova lectina lactose-específica isolada do fungo Langermannia bicolor com propriedades antibiofilme(2019-12-20) Moura, Geovanna Maria de Medeiros; Santos, Elizeu Antunes dos; ; ; Nascimento, Ermeton Duarte do; ; Rocha, Hugo Alexandre de Oliveira; ; Santos, Paula Ivani Medeiros dos;Constituted by a complex matrix of biopolymers, the structure of the bacterial biofilm acts as a strategy of resistance to environmental stress conditions, including for example host immunity and antibiotic action. This problem has intensified the search for molecules with antibiofilm action, such as those capable of interfering with the matrix structure of the biofilm and thus allowing the action of antibiotic drugs. In this context, the lectins, proteins of non-immune origin capable of binding specifically to biofilm carbohydrates, stand out. The aims of this work are isolate and characterize the lectin from the basidiomycete Langermannia bicolor (LLB) and evaluate its binding potential to bacteria and antibiofilm activity. LLB was isolated after separation with cation-exchange chromatography and gel filtration, showing specificity calcium-dependent to bind lactose, as well as high stability at different pH and temperatures values. LLB did not affect the human red blood cells and murine fibroblast strains (3T3) viability. Although LLB had no antibacterial activity, it was able to agglutinate (interaction dependent on its recognizing domain) and significantly reduce total biofilm biomass preventing early biofilm formation and preformed biofilm in Staphylococcus aureus and Pseudomonas aeruginosa. Moreover, these data suggest LLB as a future candidate for prototype antibiofilm, however further studies should be conducted to elucidate the mechanism of action as well as its possible combination with conventional antibiotics as a therapeutic strategy to reduce the development of bacterial resistance.Master Thesis Avaliação do efeito in vitro, in vivo e ex vivo do picolinato de cromo(2019-09-30) Morais, Yara Campanelli de; Rocha, Hugo Alexandre de Oliveira; ; ; Lima, Jailma Almeida de; ; Scortecci, Katia Castanho;Chromium picolinate (PICCr) is a complex made up of three picolinic acid residues (in salt form) linked by coordination to a chromium III atom. PICCr supplementation by athletes and others has been increasing every year and without a determined dose. The hypothesis of this study is the safety of prescribing varying doses of chromium picolinate. After commercial acquisition of PICCr capsules from two different countries, England and Brazil, this work began by checking by Atomic Absorption Spectrometry, the masses contained in these capsules. We found a higher concentration of the mineral in the capsules of English origin, which was selected as a sample for the research. Four cell lines: murine macrophages (RAW264.7), Chinese hamster ovary (CHO) cells, murine fibroblast (3T3) and human hepatic adecarcinoma (HepG2) cells were exposed to varying concentrations of PICCr (5 to 20 µg /mL) CHO, 3T3 and HepG2 were found to have their ability to reduce (10 to 80%) decreased MTT (3- (4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazole). RAW 264.7 cells were not affected in this analysis. HepG2 cells when exposed to PICCr (20 to 50 µg / mL) had micronucleus numbers significantly similar to cells that were not exposed to PICCr. RAW 264.7 cells showed a decrease in nitric oxide release when they were exposed to bacterial wall liposaccharides (LPS). In in vivo assays, male Swiss mice were exposed to different doses of PICCr (1000 and 2000 µg / mL) for 15 days. After this period, it was found that the treated animals had lower weight gain and decreased plasma cholesterol, triglycerides and glucose levels. They did not present macroscopic or histological changes in the hepatic and renal tissues. Treated animals were also found to have 50% decrease in serum plasma iron levels. However, metallomics analyzes show that iron and hepatic tissue iron levels have not been affected, however, human studies are necessary to confirm these observations, and may boost future studies to better understand these effects and add economic and pharmacological value to the study. PICCr.Master Thesis Polissacarídeos sulfatados da alga marrom Dictyota mertensii: avaliação do efeito antioxidante sinérgico em associação com o ácido ascórbico e purificação de uma fração(2019-09-23) Silva, Cynthia Haynara Ferreira da; Costa, Leandro Silva; Rocha, Hugo Alexandre de Oliveira; ; ; ; Filgueira, Luciana Guimarães Alves; ; Telles, Cinthia Beatrice da Silva;Seaweed are organisms with great biological diversity and wide geographicol distribution. Considering of avaliability, they are important for several areas of knowledge, such as environmental, economic and scientific. They have several organic compounds with bioactive functions, and among these are sulfated polysaccharides (SP). The brown seaweed SP (fucans and fucoidans) have several activities, including antioxidant activity, which may be enhanced by the presence of other antioxidants The antioxidant potential of PS obtained from D. mertensii is already reported in the literature. However, this is below what is found in other PS. Thus, the objective is to evaluate the synergistic effect of antioxidant activity in combination with ascorbic acid. The algal were collected SP from Potiguar coast and submitted to proteolysis and differential acetone fractionation to obtain PS, which led to obtaining 6 fractions (F0.3; F0.5; F0.7; F1.0; F1.5; F2.1). Polysaccharide fractions were subjected to chemical and physical analyzes to determine their composition and as a result, D. mertensii synthesizes structurally different PS. It was observed that the fractions are rich in heteropolysaccharides, with predominance of monosaccharides fucose, glucuronic acid and xylose in the fractions F0.3, F0.5, F1.0, F1,5 e F2.0. In addition to these monosaccharides, glucose and galactose was observed in F1.0 and F1.5. The F0.7 fraction is composed mainly of fucose and glucuronic acid. Three patterns of electrophoretic migration were observed in fractions: a slower profile was observed in fractions F0.3, F0.5 and F0.7; an intermediate profile in F1.0 and a faster profile in fractions F1.5 and F2.1. Different values were also observed for the quantification of total sugars and sulfate in fractions. A higher percentage of sugars was observed in fractions F0.7 and F0.5, with 85% and 66%, respectively, and higher sulfate percentages were observed in fractions F1.0 and F0.7, with 40.2% and 32%, respectively. Fractions were evaluated for antioxidant potential in combination with ascorbic acid in in vitro and cell culture tests. As a result, there was an increase in antioxidant potential from the synergistic effect of PS and ascorbic acid in all fractions, with emphasis on F0.7, in which potentiation of over 100% in total antioxidant capacity and 40% in reducing power activity. The analysis of cell viability in dog renal epithelial cells (MDCK) by the 3- (4,5-dimethyl-thiazol-2-yl) -2,5- diphenyltetrazolium bromide reduction assay (MTT), It was possible to observe decreased cell viability only with the use of fraction F1.5, and F1.0 and F2.0 at a concentration of 1 mg / mL The potential synergistic effect on antioxidant activity in cell culture was evaluated by the hydrogen peroxide induced oxidative stress model in MDCK cells. As a result, a protective effect was observed only with fractions F0.5, F0.7 and F1.0, evaluated separately. The F0.7 fraction was chosen for the purification steps and after the use of ultracentrifugation, ion exchange chromatography and gel filtration chromatography (HPLC) devices, the analyzes showed that F0.7 is composed of three main PS populations. The data found here open the way for further studies to propose the structure of at least one PS, as well as better understand the synergistic effect of these polysaccharides with ascorbic acid.Master Thesis Caracterização da microbiota de uma amostra de petróleo do pré-sal(2019-07-30) Freitas, Júlia Firme; Lima, Lucymara Fassarella Agnez; ; ; Porto, Ana Lúcia Figueiredo; ; Campos, Julliane Tamara Araujo de Melo; ; Dalmolin, Rodrigo Juliani Siqueira;Petroleum is one of the world's main source of energy. On Brazil pre-salt reservoir discovery and extraction has boosted oil production. However, such activity generates recurrent environmental contaminations, causing global problems. Microorganisms demonstrate the ability to degrade oil, so microbial metabolism can provide an opportunity to bioremediate contaminated environments and assist in reservoir management, contributing to maintenance oil quality or oil recovery. Identification of the microbiota in the reservoir provides data to understand the metabolism of oil degradation. It is worth mentioning that there are still no pre-salt microbiota analyzes, described in the literature, by dependent or independent culture approaches. The objective of this work was to obtain and characterize isolates and microbial consortia (indefinite and definite) capable of degrading petroleum. From a pre-salt oil sample three indefinite consortia and 26 isolates were obtained. To evaluate the potentials of the consortia and isolates, the following tests were performed: i) oil degradation ability, ii) production of biosurfactants; iii) cellular hydrophobicity; iv) biofilm production; v) enzymatic activity of hydroxylases; vi) presence of rhamnolipid and vii) growth curve. All isolates demonstrated the ability to degrade petroleum and some by-products by the DCPIP test, 46% used kerosene and 7% degraded xylene and toluene. The oil dispersion test showed that all isolates produce biosurfactants, yet 22 to emulsify kerosene. Lipase was produced by all isolates, however, only 4 strains produced esterase. The isolates were identified as belonging to the genera Bacillus (n= 9), Ochrobactrum (n = 11), Acinetobacter (n=2), Dermacoccus (n=1) e Staphylococcus (n = 2). One definite consortium was elaborated with eleven isolates that presented the best results in the functional tests. The estimation of degradation indicate that all consortia degraded 100% of oil, diesel, gasoline, soybean oil, olive oil and hexadecane, while degradation of kerosene was 46 to 90%, xylene 44 to 77% and toluene 43 to 78%; the emulsification index of the consortia ≥58%. The comparison of the results between the selected and the indefinite consortia shows similarities in the functional characteristics (growth curve, hydrocarbon degradation and biosurfactant production). It is concluded that microorganisms obtained from or enriched by petroleum have the ability to degrade oil from biofilm production and cell hydrophobicity modification, and the selected consortium presents the same potential of the native microbiota, being a good possibility for application of bioremediation; while indefinite consortia can provide data for reservoir management.Master Thesis Estudo da influência do voo com o foguete VSB-30 no transcriptoma de plantas de Saccharum spp. (cana-de-açúcar)(2017-05-26) Nascimento, Vladimir Vieira do; Scortecci, Katia Castanho; ; ; Lima, João Paulo Matos Santos; ; Calsa Júnior, Tercilio;With the submission of sugarcane plants in a flight with the VSB-30 rocket, histological and biochemical changes were observed in your different organs. Therefore, we aimed to compare their transcriptome profiles to identify in tissue samples from leaves and roots possible responses to the flight condition with the VSB-30 rocket. For this, initially, 118 Differentially Expressed Transcripts were detected with values of Log2FoldChange ≥2 or ≤-2 which were obtained by executing the DESeq package, using the blind method. Through these comparisons that were performed between the samples belonging to the groups Test flight with the VSB-30 rocket and Controls, it was verified that the highest sensitivities occurred in samples of roots tissues, which allowed to select 77 specific transcripts. In addition, the submission of the whole set of transcripts the tool STRING v.10. returned networks of orthologous proteins from the organism Sorghum bicolor. However, the analyzes of systems biology that were performed in the Cytoscape v.3.4 program, allowed to identify for the roots samples changes in Genetics information Processing, with induction, mainly, in biological processes of translation. In addition, the sample of Vertical Rocket Root showed the highest sensitivity to the microgravity and hypergravity conditions, which can be interpreted by biological systems as a condition of abiotic stress, which results in induction of the biological processes that involve the proteasome and repression for ribosome maturation. This, possibly, result in progression of the cell cycle and provides maintenance of nitrogen levels in the cell. In contrast, at tissue samples from leaves appeared as possible responses the induction of the transcription. Furthermore, on the sample Vertical Rocket Leaf, induction was detected on the metabolic pathway of glycine degradation and decarboxylation. With all that, all these results indicate the main responses to the flight with the VSB-30 rocket, which reaches microgravity and hypergravity environments, which in plants are interpreted as a condition of environmental stress.Master Thesis Caracterização da atividade antiparasitária do dietilditiocarbamato frente a diferentes cepas de Trypanosoma cruzi(2019-08-29) Oliveira, Johny Wysllas de Freitas; Silva, Marcelo de Sousa da; Rocha, Hugo Alexandre de Oliveira; ; ; ; Pontes, Daniel de Lima; ; Ribeiro, Aline Rimoldi;The Chagas Disease caused by protozoan Trypanosoma cruzi, affects thousands of people annually. The traditional pharmaceuticals used in treatment have high toxicity and are not efficient in the chronic phase of disease. Furthermore, the genetic diversity of parasite may cause different clinical manifestations and resistance for treatment. The sodium diethyldithiocarbamate attachment the dithiocarbamate class, chemical compounds of high pharmacological versatility that can interact with metals and induced the production of radical oxygen and nitrogen species. The main of this project is characterize the inhibitory action of DETC against different strains of T. cruzi. Firstly, the different strains Dm28c (TcI), Y (TcII), QMM9 (TcIII) e Cl Brener (TCVI) grown in LIT medium. The antiparasitic assay evaluated distinct concentration of DETC at time 24 hours and 48 hours, by count and resazurin methods, against epimastigote and trypomastigote forms of T. cruzi. The cellular cytotoxicity was analyzed utilizing animal lineage 3T3 (fibroblasts) and RAW (Macrophage) 24 hours after compound application and evaluated by the MTT assay. Additionally, was analyzed the parasite dead mechanisms triggered by the compound averse to T. cruzi epimastigote in 24 hours. Was observed too the mitochondrial activity of different strains of T. cruzi in epimastigote form after 24 hours of treatment by scanning electronic microscopy. In addiction, was evaluate the inhibitory action of DETC on proteolytic proteins of T. cruzi extracts by zymography followed by densitometry. Considering that the dithiocarbamate has an important inhibitory action against α-carbonic anhydrase of T. cruzi sought to demonstrate the docking of DETC in this protein through of in silico assay of molecular docking. Upon antiparasitic assay was determinate DETC concentration necessary for reducing the populational growth in 50% (IC50) each strains in 24 hours, resulting of 28,8 µM until 66,4 µM for epimastigote form and 23,5 µM until 79,0 µM for trypomastigote form depending on strain. Through of cellular cytotoxicity was evaluate inhibitory profile of compound that demonstrated moderate cytotoxicity against animal lineages 3T3 and RAW, thus determining IC50 81,46 µM for 3T3 e 96,34 µM for RAW. The inexistence or very low activity of markers annexin and PI (Inorganic phosphate),cause the unavailability determinate of cellular death mechanism by DETC against the parasite. Upon the mitochondrial potential evaluation was show the mitochondrial damages caused by DETC on different parasite strains, inhibiting the internal mitochondrial potential activity in 80%. Through the scanning electronic microscopy was observed that the compound present different actions in parasite cellular structure, resulting in membrane discontinuity, pores and ruptures. Furthermore the zymography founded the reduction in proteolytic activity of parasite extracts achieving 36%. Furthermore, DETC showed high capacity for biding with αcarbonic anhydrase of T. cruzi, analyzing the docking location in protein structure. Therefore, DETC presents like a possible candidate for alternative treatment of Chagas Disease due elevate antiparasitic activity, moderate cytotoxicity and present acting against specific targets of parasite.Master Thesis Análise do perfil epigenético de crianças com fendas labiopalatinas não sindrômicas(2019-08-14) Rêgo, Amanda Jussara Figueirêdo; Amaral, Viviane Souza do; ; ; Navoni, Julio Alejandro; ; Medeiros, Silvia Regina Batistuzzo de;Non-syndromic clefts of the lip and/or palate (NSCL/P) are birth defects arising from environmental and genetic causes. Recent studies have linked epigenetic factors, such as DNA methylation, to the inheritability of this complex malformation, as well as nutrition has been considered an environmental trigger of NSCL/P acting directly on the methylation profile. Thus, the aim of this work was to analyze the epigenetic pattern of children with NSCL/P. The dietary pattern of oral clefts children and the gestational exposure profile of their mothers were also investigated. Buccal mucosa cells were collected from all children for DNA extraction and for global and specific methylation determination. For this, the bisulfite restriction analysis (COBRA) technique was performed to the 5-methylcytosines (5-mC) of LINE-1 and Alu regions since they comprise about 30% of the human genome, in addition, the polymerase chain reaction was performed to analyze the specific methylation of BRCA1 promoter. The choice of this gene was based on its role in genome stability. In addition, the mothers collaborated answering a questionnaire about their exposure factors during pregnancy and their children dietary information. As a result, the control group showed a higher consumption of "ultraprocessed", "refined wheat", "processed vegetable fats", "simple carbohydrates" and "sausages" (p <0.05), but for the other food categories there was no statistical difference between groups. Regarding the gestation data, a low gestational programming rate was observed and consequent delay in the beginning of nutritional supplements use that support a proper embryo development. In addition, contraceptive use time prior to gestation was significantly higher for mothers in the clefts group and it was associated with a nearly two-fold increased risk for non-syndromic oral clefts (p = 0.036, 95% CI= 0.639–4.134). About the global DNA methylation, it was found a mean LINE-1 5-mC of 48% and 45.3% for control and clefts group, respectively, with significant statistical difference (p = 0.047). The BRCA1 promoter was methylated in 30.4% and 56.5% of control and clefts subjects, respectively, with no statistical difference between groups. In general, it was seen a poor quality dietary profile of the research participants and there was a higher tendency to global hypomethylation and BRCA1 hypermethylation in children with clefts. These findings corroborate with recent speculations that DNA methylation may be the result of a transgenerational epigenetic inheritance and it have effects on the etiology of this malformation. Further studies are needed to confirm these findings and to understand other epigenetic mechanisms that are acting in conjunction with the genetic susceptibility variants and as a consequence of the environmental factors responsible for this phenotype characterization.Master Thesis Caracterização química e avaliação das atividades biológicas de extratos da polpa de Melocactus zehntneri (Britton & Rose)(2019-06-26) Martins, Verônica Giuliani de Queiroz Aquino; Scortecci, Katia Castanho; Rocha, Hugo Alexandre de Oliveira; ; ; ; Hoskin, Roberta Targino; ; Santos, Deborah Yara Alves Cursino dos;In order to correlate the popular use plants and to identify new biomolecules, cacti become the aim of this study for phytochemical and pharmacological. In this context, the present work obtained six serial extracts from the cladode cactus internal part-pulpof Melocactus zehntneri and evaluated their potential antioxidants and biological through different assays using in vitro and in vivo methods. The extraction process was done using the lyophilized pulp and apolar and polar solvents, thus generating six extracts: hexane (HE), chloroform (CE), ethanol (EE), methanol (ME), final water (FWE) and only water (WE). First, sugar, protein and phenolic compounds were measured. Then, thin layer chromatography (TLC) has shown the presence of terpenes (ursolic and oleanolic acids), saponins, sugars (raffinose, fructose, sucrose and galactose), glycoproteins, as well as uronic and glucuronic acids using the descending cellulose TLC. In high performance liquid chromatography (HPLC), the preliminary results suggested the presence of phenylpropanoids, such as coumaric acids, benzoic acid, luteolin, hesperidin and ellagic acid. Moreover, the antioxidant assays were done using different concentrations (25, 50, 100, 250 and 500 μg/mL). The extracts presented an antioxidant activity at the Iron Ion Chelation, DPPH, Superoxide Ion Scavenging Assay, Hydroxyl Radical Scavenging Assay. When, it was evaluated the antibiofilm activity against the bacterium Staphylococcus epidermidis 70D MRSE, it was observed an inhibitory action in the formation of the biofilm in almost all the extracts. Moreover, it was important to observe that the aqueous extract (WE) presented the highest percentage of biofilm inhibition - 90%. Furthermore, it was evaluated if these extracts were cytotoxic for 3T3 cell line (fibroblast cells). The data showed that no cytotoxicity was observed in most of the extracts. Based on these results, it was analysed the possible protective activity from these extracts in the H2O2 presence for the 3T3 cell lines. Then, it was observed that these extracts presented a curative activity (they treated the cells). As the antioxidant activity is also associated to cell protection, then it was analysed in vitro photoprotective activity. It was observed that HE and CE extracts presented this activity, with the values of 2,4 and 3,2 respectively. Then, it was showed a potential photoprotective activity. Therefore, the results obtained showed the potential of these extracts to act as antioxidants, antibiofilm, not cytotoxicity and with curative action against H2O2 stress and activity in sunscreens, as additives.Master Thesis Efeitos da suplementação de creatina no estado redox do tecido renal de ratos diabéticos induzidos por estreptozotocina(2019-03-14) Medeiros, Matheus Anselmo; Lima, João Paulo Matos Santos; Medeiros, Karina Carla de Paula; ; ; ; Pedrosa, Lúcia de Fátima Campos; ; Bortolin, Raul Hernandes;Introduction: Creatine is a nutritional supplement that has been the subject of research in several pathologies, with a wide possibility of clinical applications. Studies have shown the effect of creatine on increasing glucose uptake by elevating the expression of GLUT4 receptors in metabolic disorders such as type II Diabetes Mellitus, however, there are no studies that report the effect of creatine on the glycemic metabolism of type I Diabetes Mellitus. Thus, this work aims to evaluate the effect of creatine supplementation in diabetic rats induced by streptozotocin. Methods: Thirty-two Wistar rats were divided into four groups: (C) animals without diabetes and without creatine supplementation (n = 8), (CCr) animals without diabetes and creatine supplementation (n = 8), (D) animals Streptozotocin-induced diabetics without supplementation (n = 8), (DCr) creatine-treated diabetic-induced diabetic animals (n = 8). Groups D and DCr received a single dose of streptozotocin (40 mg/kg i.p.). The following evaluations were performed: clinical, biochemical, as well as the analysis of redox state parameters in the renal tissue of the animals. Results and discussion: In fasting glycemia, diabetic groups presented significant hyperglycemia in comparison with the control groups, but the DCr group showed a significant improvement in relation to group D. This result corroborates previous studies that verified the same positive effect in pathological conditions that culminate in hyperglycemia. In the quantification of urea and alanine transaminase, the diabetic groups presented a significant increase in comparison with the control groups, however, the DCr group showed a significant improvement in relation to group D in both parameters. Our results of ALT and serum urea in group D can be interpreted as an increase in systemic gluconeogenesis, provable hepatic injury and protein catabolism, since urea is the main product of the ornithine cycle. The enzymatic activity of catalase, superoxide dismutase and glutathione peroxidase were significantly lower in group D compared to control groups, however, group DCr had restored activity at similar level to the control groups in all enzymes. In the quantification of the hydrogen peroxide content, group D had significantly higher levels in comparison to the control groups, however, group DCr had a similar level of restoration to the control groups in this parameter. These results converge with previous studies that observed that creatine supplementation modulates the enzymatic antioxidant activity and may be a direct antioxidant agent of reactive oxygen species. Conclusion: Creatine can improve metabolic profile in DM and restore redox state parameters in this condition. These results support creatine supplementation as a potential adjunctive therapeutic agent.Master Thesis Caracterização in silico e análise de uma sequência homóloga à sec14 em angiospermas(2019-06-07) Silva, Camila Rayane Pereira da; Scortecci, Katia Castanho; ; ; Golbert, Daiane Cristina Ferreira; ; Calsa Júnior, Tercilio;Brazil is the world's largest producer of sugarcane. Although its importance, the molecular studies are difficult due to its genomic complexity, a high ploidy level. However, the discovery of new genes is important to improve field and sugar-alcohol industry. The aim of this work was to characterize a sugarcane homologous sequence PHOSPHATIDYLINOSITOLPHOSPHATIDYLCHOLINE TRANSFER PROTEIN SEC14 that has been identified in previous studies associated to the flowering process. The in silico characterization was performed by functional analysis of the gene, construction of protein-protein interaction networks, phylogeny, three-dimensional model and molecular dynamics and co-expression analysis. In addition, a RT-qPCR expression assay was performed in different plant tissues (leaves, roots or shoot apical meristem – SAM) as well drought stress, oxidative stress and calcium sulphate treatment. The results obtained using the bioinformatics tools showed the CRAL-TRIO domain presence in the same position, suggesting a conservation of the SEC14 sequence in plants. The phylogenetic analysis indicates probable duplications of the SEC14 protein in plants. The threedimensional model showed a structural conservation due to the presence of hydrophobic pocket for lipid binding, and it was confirmed by molecular dynamics. Furthermore, it was observed in sugarcane model another domain that was not known. Expression data revealed that SEC14 gene was differentially expressed SAM in the juvenile phase from early-flowering variety and in the adult phase from late-flowering variety. Moreover, comparing the expression in leaves and roots, it was observed a lower expression in leaves. The SEC14 expression had no significant difference in leaf tissues from plants submitted to drought and oxidative stress in growth room. Furthermore, it was analyzed the potential effect of calcium treatment in sugarcane plants in field condition. Moreover, it was not observed any expression difference in leaves or in SAM. Then, the data showed here provides a better understanding of how the SEC14 gene/protein may acts on sugarcane. This data revealed new knowledge about the 3D protein structure and it also brings new perspectives for molecular studies for crops such as sugarcane.Master Thesis Efeito da suplementação com 800 UI de Alfa-Tocoferol no soro e leite de mulheres lactantes(2019-05-15) Rebouças, Amanda de Sousa; Dimenstein, Roberto; Rodrigues, Karla Danielly da Silva Ribeiro; ; ; ; Lima, João Paulo Matos Santos; ; Melo, Illana Louise Pereira de;During lactation, there is a physiological reduction of vitamin E in milk and maternal supplementation is an efficient way to increase this concentration and ensure adequate nutritional intake for the infant. However, the impact on the serum-to-breast-milk ratio is not known. Additionally, studies show that the response to supplementation occurs in a differentiated way, suggesting the possibility of being influenced both by the dose of the supplement and by other factors that may interfere with mechanisms of transfer of the vitamin to the mammary gland, as yet unknown. Therefore, the objective of the study was to evaluate the effect of maternal vitamin E supplementation on the relationship between alphatocopherol in serum and breast milk, and to investigate the factors associated with increased serum vitamin and milk. A randomized clinical trial was conducted with 79 lactating women attending a public clinic in Natal-RN, Brazil. Participants were allocated to the supplemented group, which received 800 IU of RRR-alpha-tocopherol (588 mg), and in the control group, which did not receive supplementation. Mature milk and serum were collected in both groups between 30 and 90 days postpartum (collection 1) and the next day collection 1 (collection 2). The supplemented group received supplementation immediately after collection 1. Alphatocopherol was analyzed by High Performance Liquid Chromatography and the lipid profile was determined by commercial kits and formulas. Food consumption was obtained from the 24-hour recall. To evaluate the effect of supplementation on milk, participants in the supplemented group were also divided into quartiles, according to the percentage of increase of alpha-tocopherol in milk between collection 1 and collection 2 (quartile 1 and quartiles 2- 4). There was no difference in lipid profile, dietary intake of vitamin E, concentration of alphatocopherol in milk and serum between groups in collection 1, and between collection 1 and 2 of the control group (p > 0.05), showing the homogeneity of groups. The evaluated women did not present vitamin E deficiency (SVD), but the majority presented inadequate vitamin E intake (< 16 mg). In the supplemented group there was an increase of the serum vitamin, where in the collection 1 the average was 1136.8 μg/dL and in the collection 2 was 2080.3 μg/dL (p < 0.001), as well as in the milk that went from 300.8 μg/dL to 646.3 μg/dL after supplementation (p < 0.001). After supplementation, there was a positive correlation between alpha-tocopherol in milk and serum and with dietary vitamin E. When assessing the factors associated with the effect of supplementation, only alpha-tocopherol in milk from collection 1 and food consumption of vitamin E showed a significant relation with the increase of vitamin in milk, whereas serum from collection 1 was the only one associated with positive effect of maternal serum supplementation. These findings demonstrate that in cases of high ingestion it is possible to have a higher uptake of circulating alpha-tocopherol by the mammary gland, contributing to the association between serum vitamin and breast milk. The importance of adequate intake of vitamin E from source foods to maintain satisfactory serum vitamin and milk levels is important, as well as a better efficacy of supplementation with 800 IU of alphatocopherol. This clinical trial was recorded at http://www.ensaiosclinicos.gov.br/rg/RBR38nfg2/ (RBR-38nfg2).