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Item Dieta hiperlipídica e hipernatremia: alterações autonômicas e cardiovasculares(Universidade Federal de Goiás, 2018-02-02) Amaral, Nathalia Oda; Colombari, Débora Simões de Almeida; http://lattes.cnpq.br/1684467594359403; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341; Pedrino, Gustavo Rodrigues; Oliveira, André Henrique Freiria; Gomes, Rodrigo Mello; Amaral, Vanessa Cristiane de Santana; Rebelo, Ana Cristina SilvaHypertension is the most common pathologies which affect population. Of the factors that can promote hypertension, food behavior is highlighted in relation to unhealthy eating habits present in western diet. Thus, present study sought to evaluate some aspects of this pathology in several situations, and themes were discussed separately in two chapters. In the first moment, effect of maternal hyperlipidic diet on metabolic and autonomic parameters in offspring was evaluated. It is known which obesity induced by maternal diet can modify central regulatory pathways of the fetus, mainly long-term regulation of appetite, but no work sought to evaluate the influence of the maternal hyperlipidic diet (HD) on cardiovascular and autonomic parameters in the offspring . Female Holtzman rats (280-300 g) were divided into two groups. One group received standard diet (SD) and other HD. The animals had free access to SD or HD for 6 weeks prior and during gestation and lactation period. All pups were weaned after 21 days of life and had free access to SD. After one week some offspring and mothers were submitted to glycemic test and later euthanized for removal of adipose tissue and blood. Other offspring were submitted to in situ preparation. DH increased adipose tissue in all females, but was able to change only metabolic triacylglycerols concentration. Offspring of HFD dams (OffHFD) showed an increase in adipose tissue and total cholesterol and HDL levels. Offspring of SD dams (OffSD) showed decrease in sympathetic activity after pre-colicular transection. This effect was not observed in males offspring of mothers with HFD (OffHFD). KCN infusion caused similar increases in abdominal activity (ABD), in phrenic nerve frequency (PNA f) and sympathetic activity (SNA), but in OffHFD this increase in SNA was smaller. KCN caused increase in phrenic nerve amplitude (PNA) and was higher in OffHFD. Hypercapnia resulted in increase in SNA, ABD and PNA and decrease in PNA f in both groups. The decrease in PNA f was more pronounced in OffHFD and increased PNA was higher in OffHFD. Phenylephrine caused in both groups decrease in SNA, ABD and PNA, and generated an increase in PNA f. The reduction of SNA was higher in OffHFD. These results suggest that maternal HFD during fetal development alters central connections in offspring. In second moment oxytocin effects on vascular reactivity and its role in hypernatremia-induced responses were evaluated. Changes in volume and/or extracellular compartments composition are known to evoke various autonomic, cardiovascular and hormonal responses that to modulate renal excretion of water and sodium. The main vegetative adjustments are: renal vasodilation and oxytocin secretion. Regulation of osmolarity and volume is critical for survival. Despite knowledge, no study evaluated interaction between renal sympathetic activity and oxytocin secretion on renal and cardiovascular responses induced by sodium overload. Male Wistar rats (280–350 g) were anesthetized with sodium thiopental (40 mg/kg, i.v.). Animals were also instrumented for measurement of mean arterial pressure (MAP) and renal blood flow (RBF). Renal vascular conductance (RVC) was calculated as the ratio of RBF by MAP. In anesthetized rats (n = 6), OT infusion (0.03 μg/kg, i.v.) induced renal vasodilation. Ex vivo experiments demonstrated that OT caused renal artery relaxation. Blockade of OT receptors (OTR) reduced these responses to OT, indicating a direct effect of this peptide on OTR on this artery. Hypertonic saline (3 M NaCl, 1.8 ml/kg b.wt., i.v.) was infused over 60 s. In sham rats (n=6), hypertonic saline induced renal vasodilation. The OXTR antagonist (AT; n=7) and renal denervation (RX) reduced the renal vasodilation induced by hypernatremia. Atosiban with renal denervation (RX+AT; n=7) completely abolished renal vasodilation induced by sodium overload. Intact rats excreted 51% of the injected sodium within 90 min. Natriuresis was slightly blunted by atosiban and renal denervation (42% and 42% of load, respectively), whereas atosiban with renal denervation reduced sodium excretion to 16% of the load. These results suggest that OT and renal nerves are involved in renal vasodilation and natriuresis induced by acute plasma hypernatremia. The understanding of regulatory mechanisms activated both during obesity and during hyperosmolarity allows greater possibility development new therapeutic tools for hypertension prevention and treatment.Item Marcadores bioquímicos e inflamatórios em ratos submetidos ao modelo de sobrecarga de sódio pós-natal(Universidade Federal de Goiás, 2021-09-17) Barros, Laiza Alencar Santos; Oliveira, André Henrique Freiria; http://lattes.cnpq.br/0152151142555605; Oliveira, André Henrique Freiria; Gomes, Clayson Moura; Rosa, Daniel Alves; Colugnati, Diego Basile; Almeida, Roberto Lopes deWHO data shows that the consumption of salt by the population exceeds what is indicated, which can become a public health concern, because although the ingestion of high salt levels is classically associated with the installation of hypertension, saline environment leads to an osmotic disorder which results in multiple physiological changes in the renal, central nervous and immune systems. Even studies have linked salt overload to metabolic disorders, little is known about the consequences of a hypersodium diet in the early stages of life. Experimental groups (E) of Wistar and Holtzman male rats were treated with 0.3M saline, while the control group (C) had access to water, for 60 days. During the treatment, plethysmografhy was performed for blood pressure (BP) and heart rate (HR) measurements. After a recover period, when both groups received water, biological samples were taken for analysis of hematological parameters, in blood, ad biochemical and immunological parameters, in serum and plasma. Contrary to expectations, the Holtzman rats showed no increase in BP or HR, after hypertonic saline treatment. In addition, they do not increased inflammatory cytokines, and were normal for all biochemical parameters surveyed, except for serum creatinine, with was decreased (E: 0.27 ± 0.07 vs C:0.46 ± 0.04 mg/dl, p<0.05). On the other hand, Wistar rats developed hypertension (E: 159.9 ± 5.2 mmHg vs. C: 149.7 ± 3.2 mmHg, p<0.05) and increased HR (E: 412.9 ± 7.7 bpm vs. C: 375.7 ± 12.9 bpm, p<0.05). In addition, we found differences in biochemical parameters for renal function, with elevated urea (C: 44.39 ± 0.32 mg/dl and E: 49.5 ± 0.69 mg/dl), and low creatinine (C: 0.96 ± 0.02 mg/dl and E: 068 ± 0.004 mg/dl); also differences for liver function, with increased ALT (C: 16.26 ± 0.43 mg/dl and E: 32.63 ± 0,6 mg/dl) and total and indirect bilirubin. Furthermore, components of lipidogram and serum glucose (C: 83.63 ± 0.42 mg/dl and E: 124.2 mg/dl) were elevated. Therefore, we can reinforce that there are metabolic variations between animal strains, explaining why the Holtzman rats were not sensitive to the saline protocol. In addition, changes in the biochemical parameters of Wistar rats allow us to state that sodium overload causes several metabolic diseases, with involvement of renal and liver function, besides to alterations in the metabolism of lipids and carbohydrates.Item Aplicações biotecnológicas de feijões endurecidos(Universidade Federal de Goiás, 2014-09-16) Batista, Karla de Aleluia; Bassinello, Priscila Zaczuk; Fernandes, Kátia Flávia; http://lattes.cnpq.br/9737543228759171Beans are legumes that present high content of protein, which make them one of the most important sources of nutrients in Brazil. Despite their excellent nutritional profile, changes in biochemical and physico chemical properties during storage leads to the hardening of grains, which compromise their texture and palata bility. It is known that bean’s hardening is responsible for losses about of 100.000-500.000 ton of beans per year. In addition, considering that per capita intake of beans in Brazil was 16 Kg/person/year, the content of discharged grains as function of hardening could feed 34 million of Brazilian. In this scenario, several processes have been tested aiming to provide na alternative of inclusion for these residues. In this work, hardened beans from gen us Phaseolus (Phaseolus vulgaris) and Vigna (Vigna unguiculata) were treated by autoclaving and extrusion and the products were tested regarding their applicability in biotechnological area. The extruded beans presented nutritional and technological properties that allowed their use as ingredients in bread production. Results evidenced that breads made using a mixture of extruded bean and wheat flours presented 10% higher protein and 2.5-hold higher fiber than breads made with 100% wheat flour. These results indicate that extruded bean flours constitute ingredie nts nutritionally and economically viable for technological application in breads elaboration, conferring improvement of nutritional characteristics without changing sensory attributes. The extruded hardened beans were also used as ingredient of culture me dium replacing the traditional nitrogen sources. Results evidenced that the use of extruded cowpea (Vigna unguiculata) flour is not promising since the optical density of microorganisms grown in culture medium containing extruded cowpea was 20% lower than the optical density observed in commercial media. On the other hand, the microorganisms grown in media containing extruded common bean (Phaseolus vulgaris) presented higher values of optical density and protein expression when compared to commercial broths. Alternatively, the grains were treated by autoclaving, with perspective of use the whole grain. Results evidence that after autoclaving the hardness of grains remained high, which disable their use of whole grain. However, the autoclaving process improve d the nutritional quality without modifying the technological properties. Finally, results evidenced that each genus presented a different behavior after treatment by extrusion or autoclaving. Thus, it is evident that the type and conditions processing use dwill depend of bean characteristics as well as the properties required to the proposal application.Item Atividades antimutagênica, antigenotóxica e anticitotóxica de Silybum marianum (L.) Gaertn e sua influência na expressão de genes de resposta a danos no DNA(Universidade Federal de Goiás, 2015-03-26) Borges, Flavio Fernandes Veloso; Gomes, Clever; http://lattes.cnpq.br/9545455455623006; Lee, Chen Chen; http://lattes.cnpq.br/4621907105842007; Lee, Chen Chen; Sabóia-Morais, Simone Maria Teixeira de; Reis, Paulo Roberto de Melo; Silva, Daniela de Melo e; Sousa, Neila Coelho deSilymarin (SM) is a standardized extract from the seeds and leaves of milk thistle Silybum marianum (L.) Gaertn. It is composed mainly of flavonolignans, with silibinin (SB) being its principal active constituent. Known mainly as antioxidant and hepatoprotector, SM and SB were found to be clinically effective in the treatment of a variety of liver disorders, including acute and chronic viral hepatitis, toxin and drug-induced hepatitis and cirrhosis. Due to the wide biological activities presented by SM and SB, the present study aimed to evaluate their antimutagenic activities using the Ames mutagenicity test in Salmonella typhimurium, their antigenotoxic activities using the mouse bone marrow micronucleous test and the alkaline comet assay, and to assess their effect on the gene expression pattern of some genes associated with the process of carcinogenesis and chemoprevention. To assess antimutagenicity, bacterial suspensions of Salmonella typhimurium (TA98 and TA100 strains) were treated with different concentrations of SM or SB simultaneously with the appropriate positive controls for each strain. To assess antigenotoxicity, Swiss mice were orally treated with different concentrations of SM or SB simultaneously with a single intraperitoneal dose of mitomycin C (MMC) for the micronucleus test, and human blood lymphocytes were cotreated with SM or SB and methyl methanesulfonate (MMS) for the alcaline comet assay. To investigate the role of SM and SB in modulating gene expression, we conducted microarray analysis. The results showed that SM was not significantly effective in reducing the number of frameshift mutations in strain TA98, while SB demonstrated significant protection at higher doses (p < 0.05). Regarding strain TA 100, SM and SB significantly decreased mutagenicity (point mutations) (p < 0.05). The results of the antigenotoxic evaluation demonstrated that SM and SB significantly reduced the frequency of micronucleated polychromatic erythrocytes (MNPCE) (p < 0.05). The results also indicated that SM and SB significantly attenuated MMC induced cytotoxicity (p < 0.05). In the comet assay, SM and SB significantly reduced the genotoxicity of MMS (p < 0.05), with a stronger antigenotoxic activity exerted by the extract complex (SM) than the one exerted by the isolated main active constituent (SB). The expression array analysis of five genes related to DNA damage, carcinogenesis and/or chemoprevention mechanisms demonstrated an up-regulation of PTEN and BCL2, down-regulation of BAX and ABL1 and no significant change in ETV6 expression levels.In conclusion, our results demonstrated that both SM and SB presented antimutagenic and antigenotoxic actions, as well as modulated the expression levels of genes analysed under the experimental conditions of this study.Item Farmacogenética em psiquiatria: influência dos polimorfismos CYP1A2*1F e CYP2C19*17 na refratariedade ao tratamento à clozapina e ao escitalopram(Universidade Federal de Goiás, 2015-08-26) Brito, Rodrigo Bernini de; Cruz, Aline Helena da Silva; http://lattes.cnpq.br/1273137151063484; Ghedini, Paulo César; http://lattes.cnpq.br/5789550234984454; Ghedini, Paulo César; Cruz, Aline Helena da Silva; Diniz, Denise Sisterolli; Mendonça, Helena Rezende Silva; Bicudo, Lucilene Arilho RibeiroThe aim of pharmacogenetics is to understand the hereditary basis of therapeutic response and side effects of pharmacological agents for each individual. Antipsychotics and antidepressants are effective drugs for schizophrenia and major depressive disorder (MDD) treatment, respectively. Although a number of patients respond satisfactorily to antipsychotics and antidepressants, 20-40% of them present inadequate response, and the treatment with ineffective medication may take weeks of unremitted illness, potential adverse drug reactions and nonadherence to treatment. This study aims to identify polymorphisms in genes that potentially influence the treatment response to clozapine in schizophrenic patients and the treatment with escitalopram in MDD patients. This approach involved the study of CYP1A2 and CYP2C19 genes related to the metabolism of these drugs and which may to affect the efficacy of treatment. It was studied 54 schizophrenic patients taking clozapine and 31 patients with MDD treated with escitalopram, both for long term. The investigated polymorphisms, CYP1A2*1F in schizophrenic patients and CYP2C19*2 and CYP2C19*17 in depressive patients, were analyzed by polymerase chain reaction (PCR), followed by sequencing (CYP1A2*1F) or by restriction fragment length polymorphism (RFLP) (CYP2C19*2 and *17) techniques. The results pointed for the association between CYP1A2*1F polymorphism and super-refractory clozapine treatment and for the association between CYP2C19*17 polymorphism and the decreased response to escitalopram treatment. No association was observed between CYP2C19*2 and the response to escitalopram treatment. These findings suggest that these genetic variants have an important influence on the treatment effectiveness of antipsychotics and antidepressants in psychiatric disorders, as schizophrenia and MDD. The pharmacogenetics may be useful to the psychiatrists helping in the choice of drugs and doses more efficient for each patient, reducing suffering and costs and contributing to improve the quality of life for patients and families.Item Identificação proteômica, expressão heteróloga, citolocalização, estudos de regulação transcricional e traducional da Aconitase Mitocondrial de Paracoccidioides brasiliensis(Universidade Federal de Goiás, 2009-11-18) BRITO, Wesley de Almeida; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637Paracoccidioides brasiliensis is a thermal-dimorphic fungus, the causative agent of Paracoccidioidomycosis (PCM), an important endemic mycosis in Latin America. A protein species preferentially expressed in yeast cells with a molecular mass of 80kDa and isoeletric point (pI) of 7.79 was isolated from the proteome of P. brasiliensis and characterized as an aconitase (E.C. 4.2.1.3). Aconitase is an enzyme that catalyzes the isomerization of citrate to isocitrate in both the Krebs cycle (KC) and the glyoxylate cycle (GC). We report the cloning and characterization of the cDNA encoding the aconitase of P. brasiliensis (PbACO). The cDNA showed a 2337 bp open reading frame (ORF) and encoded a predicted protein with 779 amino acids. A polyclonal antibody against the purified recombinant PbACO was obtained in order to analyze the subcellular localization of the molecule in P. brasiliensis. The protein is present in the extracellular fluid, cell wall, mitochondria, cytosol and peroxisomes of yeast cells as demonstrated by western blot and immunocytochemistry analysis. The expression analysis of the Pbaco gene was performed through quantitative real time RT-PCR and results demonstrated increasing expression during differentiation from mycelium to yeast cells. Real time RT-PCR assays was also used to evaluate the Pbaco expression when the fungus grows on media with acetate and ethanol as sole carbon sources and in different iron levels. The results demonstrated that Pbaco transcript is over expressed in acetate and ethanol as sole carbon sources and in highiron conditions.Item Avaliação das atividades genotóxica, antigenotóxica, citotóxica, anticitotóxica, angiogênica e antiangiogênica de elagitaninos utilizando ensaios in vitro e in vivo(Universidade Federal de Goiás, 2016-09-16) Carneiro, Cristiene Costa; Santos, Suzana da Costa; http://lattes.cnpq.br/7811945085200334; Lee, Chen Chen; http://lattes.cnpq.br/4621907105842007; Lee, Chen Chen; Reis, Paulo Roberto De Melo; Bailão, Elisa Flávia Luiz Cardoso; Sapanó, Mário Antônio; Cruz, Aline Helena Da SilvaPunicalagin and gemin D are ellagitannins found in some species of plants of medical importance such as Punica granatum and Geun japonicun. For this study, punicalagin and gemin D were isolated, respectively, from the leaves of Lafoensia pacari and Eugenia uniflora, two species of Brazilian medicinal plants with several biological activities, such as antitumoral, antioxidant and healing of wounds. In the present study, we evaluated the genotoxic, cytotoxic, antigenotoxic and anticytotoxic effects of gemin D using the Ames test in Salmonella typhimurium, the micronucleus (MN) test and comet assay in mice. With the punicalagin ellagitannin, we assessed the same effects mentioned above using the comet and MN tests in mice, and we also evaluated the angiogenic and antiangiogenic activities of this ellagitannin by the chick chorioallantoic membrane (CAM) angiogenic assay. The results obtained with gemin D showed that this tannin did not present genotoxic effect by the Ames and MN tests, however, in the comet assay, the highest dose of gemin D (100 mg/kg) induced increase of breaks in DNA in comparison to the negative control (p < 0.05). In the antigenotoxicity, gemin D protected DNA against the harmful action of 4-nitroquinoline-1-oxide and sodium azide by the Ames test, and also against cyclophosphamide (CPA) in pre- and co-treatment by MN and comet tests in mice, but it did not protect DNA in the post-treatment. The results obtained with punicalagin showed that this tannin exhibited no genotoxic effect by MN test and comet assay in mice. Only the highest dose of punicalagin (50 mg/kg) exhibited significant cytotoxic effect by MN test, and in the co-treatment with CPA, this cytotoxicity was enhanced. Co-treatment, pre-treatment and post-treatment of punicalagin with CPA led to a significant reduction in the number of DNA breaks and in the frequency of CPA-induced MN, indicating antigenotoxic effect. Using the CAM model, punicalagin exhibited angiogenic activity in all concentrations, mainly at the lowest concentration (12.5 µg/µL). Therefore, gemin D and punicalagin exhibited relevant antigenotoxic and cytotoxic effects, which indicate that they may be probables candidates for chemoprevention or for the development of new cancer therapies. In addition, the angiogenic activity presented by punicalagin in this study could contribute for the processes of tissue repairing and wound healing.Item Caracterização do plasmídeo pVCM 04 extraídos de Salmonella enterica isolada de carcaçãs de frangos(Universidade Federal de Goiás, 2010-05-31) CARNEIRO, Lílian Carla; BATAUS, Luiz Artur Mendes; http://lattes.cnpq.br/5637230378599476A small cryptic plasmid isolated from Salmonella enterica Enteritidis called pVCM04 was sequenced and characterizated. pVCM04 is a 3583 pb circle molecule that showed no homology with other plasmids deposited in the GenBank. 12 ORF with more than 50 aminoacids were predicted using the ORF finder program. ORF1 and ORF2 showed homology with replication proteins of different plasmids. ORF 3-5 showed homology with mobilization proteins present in several plasmids; the others seven ORF showed no homology with genes deposited in GenBank. The pVCM04 possess a region with more than 500 pb that is not associated with none of the predicted proteins. This region is organizated in a G+C rich, A+T rich and two repeat direct sequences. The second repeat direct sequence contains a region of DnaA box connection (TTTACAC). This region is probably associated to the replication origin theta type. The phylogenetic relationship among replicase and mobilization deduced protein showed highest similarity of replicase proteins than mobilization proteins. Conjugation experiments showed that the pVCM04/pUC18 fusion not have a good ability to transfer, the plasmid stability test showed that the cells lost 60% of pVCM04/pUC18 on the first day of cultivation. The characterization suggests that the pVCM04 probably would be a cryptic plasmid from fusion of different ancestral plasmid.Item Avaliação da expressão de genes de resistência às múltiplas drogas (MDRs) e de metabolização em diferentes linhagens celulares tratadas com complexos metálicos de rutênio(Universidade Federal de Goiás, 2013-02-21) Costa, Cesar Augusto Sam Tiago Vilanova; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751Não consta resumo em outro idioma.Item Estudo de associação ampla do genoma bovino para lactação ajustada em 305 dias em girolando(Universidade Federal de Goiás, 2015-10-19) Cruz, Alex Silva da; Garcia, José Fernando; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727050P5; Cruz, Aparecido Divino da; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793956J0; Minasi, Lysa Bernardes; Reis, Ângela Adamski da Silva; Taveira, Rodrigo Zaiden; Sereno, José Robson BezerraGenomic selection in a dairy cattle breeding is a new strategy in national livestock. Genome wide association study (GWAS) is known as a strategy that involve the use of molecular markers panels distributed throughout the genome which are selected for the identification of chromosomal regions that are important for the interest traits. The aim of this study was apply the GWAS strategy for 305-day milk yield in Girolando cows. We did the genotype from 404 Girolando and after quality control analysis remained 337 individuals and 45.622 markers. The GWAS analysis resulted in 52 SNPs associated to 305-day milk yield. Of these, 23 SNPs were linked to Known genes and only 3 SNPs were linked to NUB1, SLC24A2 and DGAT1 genes that already were associated with cattle lactation. The other SNPs have no relationships described in the cattle lactation literature. In addition, the milk production QTL analysis resulted in 52 SNPs and 14 genes linked or close to 1Mb of the SNP marker. The ARS-BFGL-NGS-414 SNP on BTA19 at 47.9Mbp is located near to GH1 gene. This gene is commonly accepted as causal gene for Quantitative Character Locus of milk production mainly affecting the yield in liters and solid milk components. Thus, our data suggest that NUB1 and SLC24A2 genes could be considered as candidate genes to understand the milk production in Girolando breed. Like this DGAT1 and GH1 genes are valuable predictive markers to be added to genomic selection of dairy cattle in breeding program.Item Efeitos de peptídeos derivados da beta globina LVV-H6 e LVV-H7 sobre os comportamentos tipo-ansiedade e tipo-depressão, função cardíaca e vascular de ratos(Universidade Federal de Goiás, 2019-10-11) Cruz, Kellen Rosa da; Ianzer, Danielle Alves; http://lattes.cnpq.br/7609262674053858; Custódio, Carlos Henrique Xavier; http://lattes.cnpq.br/3868498714495781; Custódio, Carlos Henrique Xavier; Ferreira, Patrícia Maria; Ghedini, Paulo Cesar; Silva, Elder Sales da; Parreira, Ricardo CambraiaHemorphins are peptides derived from the hemoglobin β-globin chain. LVV- hemorphine-6 and LVV-hemorphine-7 (LVVs) are bioactive hemorphins, which exhibit similar amino acid residue sequence, differing only by the amino acid Phenylalanine at the N-terminus of LVV-hemorphine-7. Both hemorphins reduce anxiety-like and depression-like behavior, the latter being promoted by LVV-h7, is oxytocin receptors dependent, but not the effect evoked by LVV-h6. In addition, the data in the literature about the cardiovascular effects evoked by LVV-h7 are controversial and there are no studies on the effects of LVV-h6 on this physiological system. Therefore, the objective of this study was to identify the mechanisms involved in behavioral effects and also to verify the effects on vasomotricity and cardiac function in the aorta artery and isolated heart of Wistar rats. The experimental protocols were carried out according to the norms of use of animals, under project approval by the committee of ethics and animal use of the UFG (Protocol of approval nº 090/14). The Elevated Pluz Maze (EPM) was used to evaluate the anxiety-like behavior and were then placed in the Open Field (OF) to evaluate locomotion. To evaluate the depression-like behavior, the Forced Swim test (FST) was used. The isolated vessel technique was used to evaluate vascular reactivity in thoracic aorta rings isolated and the Langendorff technique to evaluate heart function in heart i solated from Wistar rats. The anxiolytic-like effect of both hemorphins does not depend on the route of biosynthesis of catecholamines or the activation of opioid receptors. However, the antidepressant-like effect of LVVs was reversed by blockade of opioid receptors, indicating the activation of these receptors as a potential mechanism. Both LVVs similarly reduce perfusion pressure, maximal and minimum dP/dt and systolic and diastolic intraventricular pressure in heart isolated from rats, without promoting contraction or relaxation of aorta rings isolated from rats. Thus, although LVVs cause the same effects on anxiety-like and depression-like behavior, the underlying mechanisms are partially different, even with a substantial similarity in the primary structure of these hemorphins. In addition, LVVs act by decreasing cardiac function, in the evaluated parameters, in isolated heart of normotensive rats without affecting the vasomotricity of aorta rings isolated of normotensive rats.Item Desenvolvimento de materiais bioativos e biodegradáveis a partir de polissacarídeos(Universidade Federal de Goiás, 2018-10-19) Cruz, Maurício Vicente; Fernandes, Kátia Flávia; http://lattes.cnpq.br/9737543228759171; Oliveira, Sérgio Botelho de; Caramori, Samantha Salomão; Campos, Ivan Torres Nicolau de; Pereira Júnior, Marcos Antonio; Fernandes, Kátia FláviaPlastics have become indispensable materials for modern society, with a wide variety of applications, mainly in the packaging sectors such as food, pharmaceuticals, cosmetics and chemical inputs. Although they have a number of associated economic advantages, plastics are responsible for a number of environmental problems, which have reached critical levels in all countries around the world. In this context, natural polymers appear as potential substitutes for synthetic polymers, since they have a low associated environmental impact. Cashew gum polysaccharide (CGP) and starch are materials that have excellent properties for the production of film-forming matrices, which can be exploited in several biotechnological applications. In this work, the CGP was evaluated for its potential use as a support for the immobilization of cell wall degradation enzymes produced by Trichoderma asperellum T00 (TCWDE) and it’s in vitro bioactivity was evaluated for the ability to inhibit the growth of fungi Aspergillus fumigatus and Penicillium sp. In addition, CGP films in association with polyvinyl alcohol (PVA) were produced and the fractional factorial design technique was used to study the effects of composition variation on its physical, mechanical and morphological properties. The films produced by the casting technique were analyzed for total carbohydrate content, transparency (wet and dry films), thickness, maximum breaking force, tensile strength, elongation percentage, structural stability and swell index in water, sodium acetate buffer and in glycine buffer. The morphology of the films was evaluated by scanning electron microscopy. The F7 and F8 formulations were bioactivated by immobilization of trypsin inhibitor molecules: Platypodium elegans trypsin inhibitor (PeTI), Acacia polyphylla DC trypsin inhibitor (AkTI), Inga laurine trypsin inhibitor (SW.) Willd (ILTI), and talisin a lectin/trypsin inhibitor isolated from the seeds of Talisia esculenta (St. Hil.) Radlk. In addition, the starch obtained from the fruits of lobeira (Solanum lycocarpum St. Hil) was used for the production of an edible packaging destined to the food industry. The results showed that the best condition for immobilization of EDPCT’s was obtained using the 1.0 mmol L-1 metaperiodate concentration and immobilization time of 20 min with a maximal activity of immobilized N- Acetylglucosaminidase (NAGase) of 30.1 (± 0.38) U mL-1. In the optimum immobilization condition, retention of 93, 41 and 24% was obtained for the enzymes NAGase, chitinase and β-1,3- glucanase, respectively. The CGP/TCWDE system proved to be very effective in inhibiting the growth of A. fumigatus, and inhibiting the growth of Penicillium sp. to a lesser extent. The Transmission Electron Microscopy evidenced the performance of EDPCT's on the cell wall of the target fungi. The lifetime of the CGP/TCWDE was evaluated during 200 days, where a reduction of 49.6% of the initial activity was observed after 40 days of dry storage at room temperature (25 °C). After this period, no further loss in activity was detected. The fractional factorial design revealed that the CGP has positive linear effect on the remaining total carbohydrate properties, transparency and maximum breaking strength. The PVA, on the other hand, had positive linear effect on the thickness, transparency, variation of area in water and the percentage of elongation; negative linear effect on swelling in glycine buffer; and negative quadratic effect on swelling in water and glycine buffer. The sodium metaperiodate (NaIO4) presented a linear negative effect on the properties of total carbohydrates remaining, thickness, transparency and water swell index. Scanning electron microscopy (SEM) revealed that the different formulations of CGP/PVA films show differences in micropore formation capacity during swelling in water (pH 7.0), in sodium acetate buffer (pH 4, 5) and in glycine buffer (pH 9.5). Principal component analysis confirmed the distinct behavior of the formulations and their properties, revealing the existence of three main components, which together account for 91% of the analyzed properties. Immobilization tests of trypsin inhibitors by trapping demonstrated greater efficiency in the 24 h immobilization time with efficiencies of 6.7; 42.5; 3.4 and 93.4% for PeTI inhibitors, AkTI, ILTI and Talisin, respectively. The films produced from the starch of the lobeira showed high transparency, and excellent absorption capacity of dyes and food fragrances. These characteristics favor the use of this material in different segments of the food industry.Item Extração e caracterização físico-química do amido nos cormos de Trimezia juncifolia (KLATT) Benth & Hook(Universidade Federal de Goiás, 2019-08-30) Cruz, Viviane Ovidio de Almeida; Fernandes, Kátia Flávia; http://lattes.cnpq.br/9737543228759171; Fernandes, Kátia Flávia; Batista, Karla de Aleluia; Moraes, Moemy Gomes de; Leal, Maria Carolina Bezerra Di Medeiros; Cruz, Maurício VicenteThe members of Iridaceae family have the storage of carbohydrates in underground organs as main characteristic. Some Iridaceae species might store more than one type of carbohydrate, being starch the main reserve compound. Trimezia juncifolia belongs to the Iridaceae family and has a corm as an underground reserve organ and has fibrosis cataphylls, which presents a large amount of starch. This starch reserve is used to provide energy during the onset of vegetative growth and to maintain metabolism during periods of stress, such as the seasonal drought, when the plant is under the phenological stage of dormancy, interrupting its growth and reducing its metabolism. Starch is a semi-crystalline polysaccharide composed of two types of polymers: amylose and amylopectin. The starch is stored as granules and may present differences in morphology, crystalline structure, amylose and amylopectin ratio and chain size, depending on the botanical origin. In this sense, the objective of this work was to extract and characterize the starch from corms of T. juncifolia collected in the dry and wet season. Results evidenced a yield of starch extraction in the range of 57.2% in the wet season and 69.2% in the dry season. The starch granules presented a bimodal size distribution. The size of starch extracted in the dry season was higher (3.69-33.75 μm) than extracted in the wet season (2.78-14.4 μm) and both presented shape similar to wheat starch. The content of amylose was 44 % and 41 % for starch from corms collected in dry season and wet season, respectively. The birefringence of starch granules was higher in wet season. The degree of polymerization of amylopectin analyzed through chromatography showed a slight difference between starch from wet (DP 81) and dry (DP 80) season. Amylose branches showed higher difference in the degree of polymerization between wet (DP 51) and dry (DP 41) season. X-ray analysis revealed differences in starch stored in the corms. Starch from dry season presented similarity with type-A polymorph more compact whereas a C-type polymorph a mixture of type-A and type-B was observed in the wet season. The crystallinity values were 27% and 25.9% for wet and dry season starches, respectively. The endothermic transition temperature were: wet season To= 39.7 ºC, Tp= 84.1ºC, Tc= 157.4 ºC and ΔT = 117.7 ºC; dry season: To= 27.6 ºC, Tp= 73 ºC, Tc= 116.6 ºC and ΔT = 89 ºC. Furthermore, the enthalpy changes (wet season: ΔH = 84.9 J g−1 / dry season: ΔH = 54.9kJ g−1) were accessed by DCS analysis. In conclusion, the results of this study indicate that the water availability in the environment results in biochemical changes in the characteristics of the starch stored in the cataphylls from T. juncifolia.Item Produção de uma xilanase recombinante de Streptomyces sp. S27 e aplicações biotecnológicas(Universidade Federal de Goiás, 2016-12-19) Cunha, Carolina Cândida de Queiroz Brito; Bataus, Luiz Artur Mendes; http://lattes.cnpq.br/5637230378599476; Ulhoa, Cirano José; http://lattes.cnpq.br/8368469162867277; Faria, Fabricia Paula de; Georg, Raphaela de Castro; Colussi, Francieli; Steindorff, Andrei Stecca; Ulhoa, Cirano JoséThe xylan-degrading enzymes are responsible for the hydrolysis of xylan and can be produced by a variety of microorganisms such as bacteria and fungi. Bacteria produce an effective xylanolytic system composed of endoxylanase, β-xylosidases, arabinofuronosidases, glucuronidases, acetyl xylan esterase and esterase ferulic acid and have been the target of several studies on the production and application of these enzymes in biotechnological processes such as hydrolysis of different substrates to obtain fermentable sugars. Actinomycetes comprising a phylum Gram positive, aerobic, which generally form branched filaments, often in the form of mycelium. Among the actinomycetes, bacteria of the genus Streptomyces in particular, have been studied due to their important role biotechnology. In the present study, we report the expression, purification and characterization of xylanase (XynBS27) recombinantly in yeast P. pastoris. To achieve high levels of production, the gene was designed and synthesized by optimizing codon usage for P. pastoris. The synthetic gene (xynBS27) was cloned into expression vector containing the AOXI promoter followed by integration into the yeast genome. The increased production of XynBS27 was after 96 h of induction, 2% methanol at 28 °C and 200 rpm (80 U/mL), the enzyme was purified in one chromatography step molecular exclusion obtained specific activity of 8525 U/ mg its highest activity at 75 °C and pH 6, it is thermostable and showed stability at pH 5, 6 and 7. The XynBS27 has a molecular mass of 19 kDa active in zymography. Many ions were tested Al3+, NH4+, Ba2+ and Mg2+ were inducers and Ag+ ions, Hg2+ and Cu2+ inhibited, the sulfides of Mn2+, Zn2+, Fe3+ and denaturing agents EDTA and SDS inhibited XynBS27. The enzyme is tolerant to high concentrations of glucose and xylose. The Km and Vmax values were 12.38 mg/mL, 13.679 mmol/(min.mL), respectively, Ki of 180 mM competitive inhibition. The use of the enzyme in baking has proved very promising in the evaluated parameters, volume, density, reducing sugar, stiffness, consistency, firmness and water retention.Item O papel da O-glicosilação com N-acetilglucosamina na gestação(Universidade Federal de Goiás, 2021-06-14) Dela Justina, Vanessa; Vitorino, Fernanda Regina Casagrande Giachini; http://lattes.cnpq.br/3100345884689140; Vitorino, Fernanda Regina Casagrande Giachini; Priviero, Fernanda Bruschi Marinho; Passaglia, Rita de Cassia Aleixo Tostes; Filgueira, Fernando Paranaiba; Castro, Carlos Henrique deO-linked N-acetylglucosamine (O-GlcNAc) is a post-translational modification that modulates several proteins. The increase in O-Glycosylation is observed during hyperglycemia, favoring endothelial dysfunction as well as placental modifications. The hypothesis of this work was that the increase in O-Glycosylation during pregnancy, generates vascular and placental dys- function, resulting in inadequate nutritional support between the maternal -fetal interface. For this, the work was divided in two parts. PART I: To characterize how different glycemic levels affect O-GlcNAcylation of cells in different placental regions. Female Wistar rats were divided into the following groups: severe hyperglycemia (>300 mg/dL; n= 5); mild hyperglycemia (> 140 mg/dL, at least two moments in the oral glucose tolerance test; n= 7) or normoglycemia (<120 mg/dL; n= 5). Cells in the labyrinth and junctional zones are targeted by the accumu- lation of O-GlcNAc in response to severe hyperglycemia, possibly due to reduced O-GlcNAcase (OGA) expression. O-GlcNAc is not seen during mild hyperglycemia, possibly due to OGA compensatory expression/activity. In addition, morphometric changes, occurring simultane- ously with increased accumulation of O-GlcNAc in placental tissue, can contribute to placental dysfunction during hyperglycemia. PART II:) To assess whether the increase in O-GlcNAc impairs the endothelial function of the uterine artery. Pregnant (P) or non-pregnant (NP) rats were treated with glucosamine (300mg/kg) or vehicle for 21 days, i.p. Glucosamine treatment increased the expression of O-GlcNAc in UA in NP rats. A decrease in endothelium-dependent relaxation was observed in UA of treated NP rats, compared to the vehicle, which was abo- lished by incubation of the nitric oxide synthase inhibitor (eNOS). eNOS activity as well as total Akt expression are reduced after treatment with glucosamine in NP rats. In addition, UAs from NP rats treated with glucosamine showed an increase in the activation of glycogen sy n- thase kinase 3 beta (GSK3β), as well as an increase in the expression of OGT. Interestingly, during pregnancy, treatment with glucosamine decreases the expression of O-GlcNAc in UA, accompanied by an improvement in relaxation for acetylcholine, while the activities of eNOS and GSK3β and the total expressions of Akt and OGT remained unchanged. Endothelium- independent relaxation was not altered in the groups studied. Therefore, the underlying me- chanism that occurs between P and NP is, at least in part, dependent on Akt/GSK3β/OGT mo- dification. We believe that, during pregnancy, hormonal changes play a vascular protective role, preventing endothelial dysfunction induced by O-glycosylation. In the future, it will be interesting to assess whether strategies that normalize O-GlcNAcylation levels in placental proteins will result in gestational success in pathological conditions, including hyperglycemia.Item Avaliação neurofarmacológica das atividades tipo ansiolítica e/ou antidepressiva da fração diclorometano, ácido oleanólico e (E)-metilisoeugenol das folhas de pimenta pseudocaryophyllus (Gomes) L. R. Landrum (Myrtaceae) quimiotipo (E)-metilisoeugenol(Universidade Federal de Goiás, 2015-10-16) Fajemiroye, James Oluwagbamigbe; Costa, Elson Alves; http://lattes.cnpq.br/2607893423583912; Costa, Elson Alves; http://lattes.cnpq.br/2607893423583912; Rocha, Fábio Fagundes da; Paula, Joelma Abadia Marciano de; Ghedini, Paulo César; Custódio, Carlos Henrique XavierDepression and anxiety are widely acclaimed as psychiatric disorders of global concern. These disorders are among the leading causes of disability worldwide. Unsatisfactory responses of patients to the available pharmacotherapy make the search for new drugs a necessity. Medicinal plants remain important source of new drugs and new chemical entities. The ethnopharmacological knowledge and previous data have revealed calming and anxiolytic like effects of the organic leaf extract of Pimenta pseudocaryophyllus (Gomes) L.R. Landrum. The present study sought to investigate antidepressive like effect of dichloromethane fraction (DF) of the ethanolic leaf extract of Pimenta pseudocaryophyllus as well as anxiolytic and antidepressive like effects of oleanolic acid (OA), (E) methyl isoeugenol (MIE) and possible mechanisms of action that are involved. Animal models like barbiturate-induced sleep, light dark box test (LDB), elevated plusmaze (EPM), open field (OF), wire hanging test, pentylenetetrazol-induced convulsion test, forced swimming test (FST), tail suspension test (TST) were conducted to evaluate behavioural alterations that were elicited by the administrations of vehicle, DF, OA, MIE or reference drugs. Bioassays (ex vivo and in vitro) of monoamine oxidase (MAO) and quantification of hippocampal level of brain derived neurotrophic factor (BDNF) were conducted in an attempt to elucidate possible mechanisms of action. Oral administration of DF 125, 250 or 500 mg/kg (potentiated the hypnotic effect of sodium pentobarbital). In the TST and FST, DF 125 or 250 mg/kg induced antidepressant-like response. The data obtained in the OF suggest sedative effect of DF at 500 mg/kg. Pretreatment (i.p) with pchlorophenylalanine methyl ester (PCPA) 100 mg/kg (serotonin depletor) or 𝛼-methyl-ptyrosine (AMPT) 100 mg/kg (catecholamine depletor) blocked anti-immobility effect of DF viii in the FST. The enzymatic activity of MAO remained unaltered by DF. Oral administration of OA (5-20 mg/kg) increased the duration of barbiturate - induced sleep and demonstrated anxiolytic like effect in both LDB and EPM. In the FST and TST, OA 5-20 mg/kg elicited antidepressant like effect without altering locomotion activity of the animals. The antidepressant like effect of OA was attenuated by NAN-190 (non-selective antagonist of 5-HT1A), AMPT, PCPA, WAY and PRAZ. Chronic administration of OA increased hippocampal level of BDNF. Oral administration of MIE 250 or 500 mg/kg potentiated hypnotic effect of sodium pentobarbital without protecting mice against PTZ - induced convulsion. The parameters evaluated in the LDB, EPM and OF demonstrated anxiolytic like property of MIE. This effect was blocked by WAY (selective antagonist of 5-HT1A) pretreatment. MIE 125 or 250 mg/kg showed antidepressant like effect in the FST. Locomotion activity of the animal in the OF remained unaltered by MIE administration at 125 or 250 mg/kg. Pretreatment of mice with PCPA attenuated antidepressant like property of MIE. In conclusion, our findings demonstrated anxiolytic and/or antidepressant like effects of dichloromethane fraction, oleanolic acid and (E) methyl isoeugenol, thereby suggesting the involvement of monoaminergic pathway.Item Morfologia, taxonomia, filogenia, anatomia foliar e titoquímica de espécies do gênero Hyptis Jacq. (Labiatae) ocorrentes em em Goiás e Tocantins(Universidade Federal de Goiás, 2010-04-29) FERREIRA, Heleno Dias; FERRI, Pedro Henrique; http://lattes.cnpq.br/2129799749473005; REZENDE, Maria Helena; http://lattes.cnpq.br/5093753722360659The genus Hyptis, with 280 species worldwide, belongs to the family Labiatae (Lamiaceae), subfamily Nepetoideae, tribe Ocimae, and subtribe Hyptidinae. The genus was divided in 27 sections, with 13 of them (Apodotes, Cephalohyptis, Cyanocephalanthus, Cyrta, Eriosphaeria, Gymneia, Induratae, Mesosphaeria, Polydesmia, Pusilae, Trichosphaeria, and Hylodontes) occur in the states of Goiás and Tocantins (Brazil). The collection of botanical material for morphological, taxonomical, anatomical, phytochemical studies, as well as geographical distribution and phylogenetic relationships, was realized with several field trips in the states of Goiás and Tocantins. For morphological studies, several loans of herbarium specimens of Hyptis were requested from Brazilian and international institutions. The genus Hyptis is mostly American, with Neotropical distribution, ranging from southern United States to Argentina. In South America, most species of Hyptis are associated with open areas, and occur from 300 m altitude in the valley of the Rio Araguaia, in Goiás, to 3300 m in the western slopes of the Peruvian Andes. In Goiás and Tocantins they can be found in several vegetation types: cerrado, rocky outcrops, margins of cow pastures, cultivated fields, wet fields, humid forests, and flooded areas with Mauritia palms. In these states they occur in a variety of soils: sandy, sandygravelly, with rocky outcrops, hydromorphic, and latossoils. Specie of the section Cyrta are exclusive of humid and flooded soils. The two main centers of diversity of Hyptis in Brazil are in the states of Minas Gerais and Goiás. Many herbarium specimens were analyzed and many species were studied and collected during field trips in Goiás and Tocantins. The geographic position of each collection was verified with the use of a GPS instrument. The specimens collected were deposited in the UFG Herbarium of the Federal University of Goiás. Thirty-two distribution maps of Hyptis in Goiás and Tocantins were elaborated from the specimens studied. A total of 89 species was catalogued, and 18 of them reported from Goiás and Tocantins were not found during the collecting trips. A key for the identification of the species was elaborated using morphological characters. A parsimony analysis using 35 morphological characters was realized, obtaining 1864 most parsimonious trees and a strict consensus tree. The parsimony analysis supports the monophyly of the sections Gymneya, Cyrta, Apodotes, and Cyanocephalus (except for the H. nitidula - H. peduncularis clade), and indicates that the sections Mesospheria, Xylodontes, Hyptis, Polydesmia, and Eriosphaeria are not monophyletic. Section Trichosphaeria is not well resolved. An anatomical study of 60 species, representing 11 sections of Hyptis, was undertaken. Some characters, as hypodermis, schlerenchimatic sheath extensions, trichomes, stomatal cripts, supply useful taxonomic for inferring phylogenetic relationships within the genus. For the analyses of chemical components, 29 species and two varieties of Hyptis were selected. With the chemical analises, 216 constituents of essential oils were identified, mostly monotherpenes and sesquiterpenes. Average percentage and pattern of each chemical component were calculated. The most common chemical components were arbitrarily selected, as present in nine or more species: sabinene (9 spp.), 1, alpha-tujene, alpha-cubebene, beta-selinene, and 14-hidroxy-(Z)-cariophyllene (10 spp.). alpha-muurolol and 8(13)-dien- 5beta-ol (11 spp.), mircene, cariophil-4 and germacrene B (12 spp.), gamma-cadinene, epialpha- caninol (13 spp.) and 14-hydroxy-9-epi-(E)-cariofilene (13 spp.), 1-epi-cubenol- and gamma-muurolene (14 spp.), humulene poxide II and bicyclogermacrene (16 spp.), betaelemene and alpha-cadinol (17 spp.), beta-pinene (18 spp.), limonene (19 spp.), espatulenol and germacrene D (24 spp.), alpha copaene and beta-bourbonene (25 spp.), delta-cadinene (26 spp.), alpha-humulene (27 spp.), cariofilene oxide and (E)-cariofilene (28 spp.). Two dendrograms were obtained with two multivariate analyses of chemical constituents, one with quantitative data and the other with qualitative data (presence/absence). Species of section Cyanocephalus were the only ones found in a consistent group in both dendrograms based on chemical constituents. The data obtained in the present study contribute important information for the taxonomy and phylogenetic relationships of the genus Hyptis and the subtribe Hyptidinae, and ultimately for the family Labiatae.Item Biotividade de extratos e frações das folhas da Eugenia uniflora L. e da Hyptidendron canum (Pohl ex Benth.) Harley em microrganismos (Bactérias e fungo) e em Oreochromis nilotius(Universidade Federal de Goiás, 2009-01-22) FIUZA, Tatiana de Sousa; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128; SABÓIA-MORAIS, Simone Maria Teixeira de; http://lattes.cnpq.br/6723881044959716The purpose of this work is to perform the pharmacognostic studies of the species Hyptidendron canum (Pohl ex Benth.) Harley (Lamiaceae) and Eugenia uniflora L. (Myrtaceae), investigate the population variability of the constituents of the essential oil from H. canum leaves and inflorescences, and assess the bioactivity of the crude ethanol extracts and fractions from leaves of this plants against microorganisms (bacteria and the fungus Candida albicans) and on Oreochromis niloticus Linnaeus. For the variability study of the H. canum essential oil, samples obtained from leaves and inflorescences from Hidrolândia, Silvânia, Bela Vista and Goiânia cities were analyzed by gas chromatography and gas chromatography with mass spectrometry. The anatomic analysis and the phytochemical screening of the leaves from the two species were performed using conventional techniques. The antimicrobial activity was assessed with Gram-positive and Gram-negative bacteria and with the Candida albicans fungus using the well diffusion test and the agar dilution method to determine the minimal inhibitory concentration (MIC). The biological activities of the crude ethanol extract and ethyl acetate, hexanic and chloroformic fractions of the leaves from the two species were tested in O. niloticus fish hepatopancreas and gill. The H. canum essential oils analysis indicated high chemovariability in the oils from different locations. The phytochemical screening and the thin layer chromatography (TLC) analysis of the H. canum leaves evidenced the presence of flavonoids, saponins, terpenes and lignans, while the E. uniflora leaves evidenced the presence of tannins, steroids, triterpenes, anthraquinonic heterosides, saponinic and flavonoids. As for the antimicrobial activity testing, the crude ethanol extracts from both species presented antimicrobial activity against all the microorganisms tested and the E. uniflora ethyl acetate fraction presented satisfactory activity against resistant Pseudomonas aeruginosa. The H. canum crude extract and fractions presented vasoactive activity in the hepatopancreas and gill of the tilapias, causing varying degrees of vasodilation and vascular congestion. An apparent proliferation of blood capillaries was detected in the hepatopancreas of the fish that ingested the hexanic fraction. The E. uniflora crude extract and the ethyl acetate, chloroform and hexanic fractions caused vasodilation, vascular congestion and tissue alterations in the O. niloticus hepatopancreas and gillsItem Caracterização do estado indiferenciado de células tronco embrionárias murinas expandidas na presença de nanopartículas magnéticas e isolamento de células tronco embrionárias a partir de blastócitos bovinos(Universidade Federal de Goiás, 2009-08-14) FREITAS, Erika Regina Leal de; GUILLO, Lídia Andreu; http://lattes.cnpq.br/3401436781775091Magnetic nanoparticles (MNPs) have been used in a great variety of biomedical applications, especially in cancer treatment, drug delivery, and diagnosis by magnetic resonance imaging. Embryonic stem cells (ESCs), due to its capacity of auto-renewal and differentiation in some types of cells, offer a great potential for its use in tissue regeneration and in alternative treatments for many degenerative diseases. The study had as aims: i) to evaluate in vitro citotoxicity of maghemite nanoparticles functionalized with lauric acid, DMSA, and citrate in human melanoma cells (SK-MEL-37) by the MTT assay, electronic microscopy, and DNA electroforesis by agarose gel; ii) to develop a culture system using the previously selected MNPs and a magnet to expand in vitro murine embryonic stem cells (mESCs) in the absence of a co-culture of murine embryonic fibroblasts (MEF) (the indifferentiated state of the mES was analyzed by alkaline phosphatase cytochemistry, electronic microscopy, and analysis of Oct-4 and Nanog gene expression by RT-PCR); iii) to isolate and expand ESCs from bovine blastocysts, and to characterize its pluripotency by analysis of Oct-4 and STAT-3 gene expression by RT-PCR. The MNPs coated with lauric acid, citrate, and DMSA showed no citotoxicity, judging by the high values of IC50 found (254, 433 and 2260 μg-iron/ml, respectively), and that the nanoparticles coated with citrate was chosen to expand the mESCs. The doubling time for the cells cultivated in the presence of MNPs was slightly higher than in the presence of MEF (20.67 ± 0.19 vs. 15.95 ± 0.21) (p= 0.001). The mESCs cultivated in the presence of MNPs showed morphology similar to ESCs, and its pluripotency was confirmed by expression of indifferentiation markers Oct-4 and Nanog by RT-PCR and high alkaline phosphatase activity. One bovine embryonic stem cell (bESCs) line was obtained and maintained by six subcultures for 60 days period. The pluripotency of the bESCs was confirmed morphologically as well as by Oct-4 e STAT-3 gene expressionItem Caracterização genética da população do estado de Goiás baseada em marcadores STRs autossômicos e do cromossomo Y(Universidade Federal de Goiás, 2013-12-17) Gigonzac, Thaís Cidália Vieira; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985; Cruz, Aparecido Divino da; Silva, Cláudio Carlos; Vasconcelos, Breno de Faria e; Rodrigues, Flávia Melo; Georg, Raphaela de Castro(Sem resumo em outra língua)