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Item Diversidade, relações filogenéticas e perfil de expressão gênica de transportadores MFS em Trichoderma spp(Universidade Federal de Goiás, 2021-04-21) Barbosa Filho, Jomal Rodrigues; Georg, Raphaela de Castro; http://lattes.cnpq.br/3971276154067590; Georg, Raphaela de Castro; Coelho, Alexandre Siqueira Guedes; Steindorff, Andrei SteccaThe Trichoderma genus comprises filamentous fungi with mycoparasitism ability and high tolerance to abiotic stresses, such as the presence of metals. In both conditions, cellular transporters are vital to nutrients absorption and metallic ion extrusion. Among those, the Major Facilitator Superfamily (MFS) correspond to the largest superfamily of secondary transporters found in every organism, however a deep understanding of those is needed in Trichoderma spp. The present work aimed to identify the MFS transporters from 32 species of this genus, as well as to study their phylogenetic relationship and gene expression profiles. Global MFS identification were performed in silico using the Trichoderma spp. reference proteomes available on the Mycocosm database from Joint Genome Institute (JGI), by searching for annotated aminoacid sequences using the BlastP tool. The sequences were categorized in their respective families, according to reference sequences from the Transporter Classification Database (TCDB), and maximum likelihood phylogenetic trees were estimated for each of the families, using the IQTREE software. Lastly, MFS genes expression profiles were investigated during mycoparasitism and presence of metals, using RNA-Seq public data. 8,467 putative MFS were identified in Trichoderma spp., with a mean of 264 ± 51 sequences, and this number was higher than those found in other organisms. Mycoparasitic species presented a higher number of MFS. 55.5% of the sequences showed 12 transmembrane helices and 85.5% showed molecular weights between 50 and 70 kDa, with a mean of 58 kDa. In total, 24 MFS families were identified in the analyzed species, and the families regarding to nutrition, maintenance of homeostasis and cell defense showed a higher number of members. We did not observed enrichment of families in any species analyzed. In the phylogenetic inference the sequences grouped according to their functions and not to their lineage, moreover, SHS and GPH families showed subdivisions. In T. harzianum, besides a higher number of differently expressed genes during mycoparasitism, a higher number of upregulated genes was also observed in this condition compared to those in the presence of metals. In the presence of cadmium, we observed a higher downregulation of MFS genes than in the presence of aluminum. We also observed that ACS and DHA2 families were mostly differentially expressed during mycoparasitism 24 h, whilst FHS and PHS were mostly differentially expressed during the presence of aluminum 1.5 and 3.0 mg/mL, and MCT during the presence of cadmium 2.0 mg/mL. This work shed light on the description of MFS transporters and their functions during some of the biotic and abiotic stresses faced by Trichoderma spp. in the course of its development on environment.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 Crisina previne a neurotoxicidade induzida por cloreto de alumínio em camundongos swiss machos(Universidade Federal de Goiás, 2022-01-28) Campos, Hericles Mesquita; Ghedini, Paulo César; http://lattes.cnpq.br/5789550234984454; Ghedini, Paulo César; Duarte, Djane Braz; Munhoz, Carolina DemarchiChronic aluminium (Al) metal accumulation contributes to the progression of several neurological and neurodegenerative diseases. In particular, Al promotes oxidative damage, neuronal death, and functional decline resulting in deficits in cognition, memory and behavior. In this regard, the use of natural antioxidants can be a strategy to prevent the neurotoxicity of Al. Chrysin is a natural phenolic compound found in foods such as honey, passion fruit, and propolis and with antioxidant and anti-inflammatory properties. Here, we evaluated the chrysin neuroprotective effects against the neurotoxicity induced by chronic exposure to aluminium chloride (AlCl3), in mice (100 mg/kg, p.o.). The experimental protocol consisted of the treatment of mice with AlCl3 for ninety consecutive days, being the chrysin administration (10, 30. 100 mg/kg, p.o.) started on the forty-sixth day of the Al intake. Chrysin reduced the cognitive impairment induced by AlCl3 in the step-down passive avoidance task, normalizing the hippocampus acetylcholinesterase and butyrylcholinesterase activities. Along with this, chrysin decreased oxidative damage observed in the parameters of the lipid peroxidation, protein carbonylation, catalase and superoxide dismutase activities in the brain cortex and hippocampus. Furthermore, necrotic cells frequency was also decreased by chrysin in the same brain regions. These results highlighted that chrysin is able to reduce the neurotoxic effects associated with Al chronic exposure, being a potential compound present in foods that can be used for the benefit of mental health.Item Efeito antipsicótico de novos inibidores do transportador de prolina (SLC6A7)(Universidade Federal de Goiás, 2020-05-29) Carvalho, Gustavo Almeida de; Pinto, Mauro Cunha Xavier; http://lattes.cnpq.br/0868250984727943; Pinto, Mauro Cunha Xavier; Menegatti, Ricardo; Leite, Jacqueline AlvesSchizophrenia is a serious psychiatric disorder that expresses a complex set of positive, negative and/or cognitive symptoms. Some hypotheses attempt to explain the cause of schizophrenia, one of which is the glutamatergic hypothesis based on pharmacological evidence and pathophysiological studies that point to a hypofunction of NMDA receptor signaling in the brain. Proline transporters (SLC6A7) are predominantly expressed in the central nervous system (CNS) and are associated with glutamatergic neurotransmission, however, their role in regulating neural function and pharmacological potential is little known. The objective of this work was to pharmacologically characterize new prototypes of proline transporter inhibitors (SLC6A7) and to test the effect of one of these in a ketamine-induced psychosis model. The results presented here demonstrate that the compounds LQFM 215, LQFM 216 and LQFM 217 have a low impact on the viability of LUHMES cells and in culture of peripheral neurons and are capable of reducing the uptake of intracellular proline in synaptosomes. The compound LQFM 215 had a greater impact on the viability of LUHMES cells as well as on peripheral neurons and a greater inhibitory effect on the uptake of proline. In the behavior of mice, the compound LQFM 215 reduced the mobility of the animals, without prejudice to the working memory in the highest tested dose of 10 mg/kg and was not able to induce anxious or depressive behaviors. In the marble burying test the compound LQFM 215 in all the tested doses of 2.5, 5 and 10 mg/kg was able to stimulate this behavior. By testing the compound LQFM 215 in the ketamine-induced psychosis model, he was able to reduce the animals' induced hyperlocomotion at the tested doses of 20 and 30 mg/kg. The set of these results indicates that the compound LQFM 215 is a neuroactive compound with an effect on the mobility of animals and with antipsychotic potential. This work demonstrates that PROT inhibitors can be targets for the development of new antipsychotics.Item Estimulação adrenérgica causada pela administração de isoproterenol induz disfunção metabólica e remodelamento pancreático em ratos Wistar(Universidade Federal de Goiás, 2022-08-30) Castro, Alessandra Gisele de; Gomes, Rodrigo Mello; http://lattes.cnpq.br/3121095341590269; Gomes, Rodrigo Mello; Mascioli, Cristina da Costa Krewer; Custódio, Carlos Henrique XavierIsoproterenol is a drug commonly used for the treatment of bradycardia, heart block, bronchospasm during anesthesia and rarely for asthma. It is a non-selective β-adrenergic agonist analogue of epinephrine, for this reason it promotes positive inotropic and chronotropic effects in cardiac tissue, being used as an experimental model for the study of cardiac alterations. It is known that the adrenergic stimulation caused by the administration of isoproterenol induces cardiac remodeling, usually through pathways mainly related to redox status and inflammatory cytokines. However, little is known about the metabolic damage of this model. Thus, this study aims to evaluate the effects of isoproterenol administration on metabolic and morphological parameters. Adult Wistar rats (50-60 days old; weighing 200- 250g) were allocated into two experimental groups: control rats (CO) that received vehicle (150 mM NaCl, 0.1ml/kg/day; i.p.), and rats that received received injections of isoproterenol (ISO; 1 mg/kg/day; i.p.) for 7 consecutive days. After the treatment period, the animals were submitted to in vivo tests and later euthanized for sample collection. The animals in the ISO group did not show a significant difference in body mass compared to the CO animals. On the other hand, ISO animals showed increased brown adipose tissue mass and brown adipose area. Furthermore, the administration of isoproterenol promoted pancreatic remodeling, with an increase in the area of pancreatic islets and interstitial fibrosis. In short, in addition to cardiac changes, the adrenergic overload induced by isoproterenol administration was also able to promote significant morphological changes in brown adipose tissue and pancreas, these changes may be linked to changes in the heart and in other systems, thus, the study Integrated physiological training is fundamental for a better understanding of the exacerbated stimulation of β-adrenergic receptors.Item Efeitos da restrição alimentar severa e moderada durante a lactação sobre parâmetros metabólicos e morfológicos maternais e parâmetros cardiovasculares na prole na idade adulta(Universidade Federal de Goiás, 2022-07-26) Cavalcante, Keilah Valéria Naves; Gomes, Rodrigo Mello; http://lattes.cnpq.br/3121095341590269; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341; Pedrino, Gustavo Rodrigues; Costa, Renata Mazaro; Ferreira, Patricia MariaDiseases developed in adulthood may be associated with conditions to which the individual was exposed in the early stages of life. It has long been postulated that changes in fetal development can trigger different effects on the newborn's quality of life. However, the great responsiveness to environmental/nutritional stimuli in postnatal phases such as lactation and puberty has been reported in the literature as an important factor in long-term phenotypic adaptation. Lactation is an important period of development, mainly due to leptin arises where the neuroendocrine circuitry is consolidated, so breast milk is the main source of nutrients for puppies in this growth phase. The objective of this study was to evaluate the effects caused on mothers due to food intervention, and the effects on cardiovascular parameters in the offspring, of both sexes, of mothers submitted to malnutrition during lactation. Pregnant Wistar rats were allocated to one of three experimental groups: Control (CO) to which a standard diet was offered ad libitum; Moderate Food Restriction (MFR, Moderate Food Restriction) for which a diet restricted to 50% of the daily consumption of the CO mothers was offered; and Severe Food Restriction (SFR) for which a low-protein diet (4.5 g protein / 100 g ration) was offered ad libitum. To maintain equity between litters, after calving all litters were adjusted to 9 pups (balancing the numbers of males and females). Feeding interventions were performed from the 1st to the 14th day of lactation, and from the 14th to the 21st day of lactation all dams received a standard diet ad libitum. The offspring of all groups were fed a standard rodent diet ad libitum throughout the experimental period. All animals were offered filtered water ad libitum. During the food intervention, the body mass and food intake of the mothers were monitored, and after weaning, samples were collected for serum biochemical measurements (Total Proteins, Albumin, Total Cholesterol, Triglycerides, HDL, TGO, Alkaline Phosphatase and Glucose) and breast milk. (Total proteins and Triglycerides), and histological analyzes (diameter of cardiomyocytes and thickness of the aorta). The offspring were followed up until 120 days of age, when the echocardiogram, invasive blood pressure recording, baroreflex test and vascular reactivity were performed. morphometric evaluation and sample collection. The protocols were approved by the UFG ethics committee (case number 023/2015). MFR mothers had reduced caloric and macronutrient intake due to the 50% food restriction protocol and had reduced ovarian adipose tissue weight. The SFR mothers showed a reduction in food intake, and consequently, there was a reduction in caloric and macronutrient intake, and a decrease in heart mass and an increase in brown adipose tissue. Males in the MFR group showed cardiovascular changes, such as increased blood pressure (BP), impaired baroreceptor sensitivity, cardiac remodeling, and changes in vascular reactivity. In contrast, males in the SFR group did not show changes in BP and cardiac tissue, however there was a loss in the sensitivity of the baroreceptors. Females from both food restriction groups did not show any cardiovascular changes, which indicates a cardioprotective effect when compared to their opposite sex pairs, possibly this cardioprotection is conferred by estrogen. Based on the results found, we conclude that moderate food restriction during lactation promotes functional and morphological cardiovascular changes in males in adulthood.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 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 Análise de diferentes misturas enzimáticas do fungo Humicola grisea var. thermoidea na hidrólise da fração de hemicelulose de bagaço de cana-de-açúcar(Universidade Federal de Goiás, 2011-08-31) Faria, Syd Pereira; Castro, Carlos Henrique de; http://lattes.cnpq.br/6354834854727314; Faria, Fabrícia Paula de; http://lattes.cnpq.br/3739169267521003; Faria, Fabrícia Paula de; Lopes, Francis Júlio Fagundes; Ulhoa, Cirano JoséThe biomass consists mainly of cellulose, hemicellulose and lignin. Cellulose is the most abundant polymer xylan and the main component of hemicellulose. Enzymatic hydrolysis is an important step in the bioconversion of cellulose and hemicellulose fraction of lignocellulosic residues. However, for the conversion of the hemicellulose, the substrate must undergo a pretreatment step which allows the refining of biomass at the microscopic level, separating the same into its main components (cellulose, hemicellulose and lignin) or loosening the fibers of its components, facilitating the accessibility of enzymes to hemicellulose chain. The conversion of cellulose and xylan to glucose and xylose may be performed by a group of enzymes produced by bacteria and fungi. The thermophilic fungus Humicola grisea var thermoidea produces an efficient complex of cellulolytic enzymes (endoglucanases, cellobiohydrolases and β-glucosidase) and xylanolítics (endoxylanases and β-xylosidase) with high thermal stability when cultivated in different lignocellulosic substrates. The aim of this study was to analyze the efficiency of enzymes produced by H.grisea in the hydrolysis of the hemicellulose fraction of sugarcane bagasse (BCA). BCA assays used in the hydrolysis was subjected to alkaline pretreatment (Treatment I, II and III) and steam explosion. BCA pretreated was subsequently hydrolyzed with different enzyme mixtures using culture supernatant of the fungus H. grisea supplemented with recombinant enzymes endoxylanase (HXYN2r), cellobiohydrolase (CBH1.2r) and β-xylosidase (XYNB2r). The results showed that the pre-treatment was most suitable to steam explosion which revealed a yield of about 78% xylan. The best enzyme formulation was a mixture of the supernatant of culture of H. grisea (SHG) grown in wheat bran (WB) and BCA supplemented with HXYN2r (950 U), CBH1.2r (950 U) and XYNB2r (1.17 U).Item Efeito transgeracional da obesidade materna por superalimentação precoce sobre os parâmetros biométricos da prole na vida adulta(Universidade Federal de Goiás, 2022-08-29) Ferreira, Lucas Araújo; Gomes, Rodrigo Mello; http://lattes.cnpq.br/3121095341590269; Gomes, Rodrigo Mello; Miranda, Rosiane Aparecida; Costa, Renata Mazaro eAmong the most prevalent non-communicable diseases today, obesity stands out at increasing levels each year. More recent data from the World Health Organization (WHO) indicate that about 650 million adults are obese, that is, many of these individuals are of reproductive age. Based on this information, the present study aims to evaluate the effects of obesity induced by the maternal postnatal overfeeding model on metabolic parameters in the offspring of Wistar rats in adulthood. For this, non-obese Wistar rats with 70 days of age were placed to cross with non-obese rats, constituting the F0 generation. Three days after conception of the first generation (F1), litter reduction was carried out in the proportion of 3 female animals per litter constituted by the reduced litter group (Small Litter, SL) and 9 female animals per litter for the control groups (Normal Litter, NL). At 70 days of age, F1 rats were crossed with non-obese rats and after conception of the second generation (F2), litters were standardized to 6 pups per mother (3 male pups and 3 female pups). After weaning, the F2 generation was separated into the following experimental groups: male and female offspring from NL mothers: M-NLO and F-NLO; and pups from SL mothers: M-SLO and F-SLO. The pups were followed up to 120 days of age, when euthanasia and sample collection were performed. Both M-SLO and F-SLO offspring had higher food intake and body mass gain throughout the experimental period, presented obese phenotype, increased pancreas mass and pancreatic islet area. However, biochemical analyzes revealed that only the M-SLO offspring had an increase in plasma concentrations of glucose and triglycerides. Molecular analyzes also showed that only the M-SLO offspring had a reduction in the expression of PI3K and p-AKT/AKT ratio in the hypothalamus. Taken together, our data evidence that postnatal maternal overfeeding induces an obese phenotype in both male and female offspring during adulthood, as well as pancreatic islet remodeling. However, only in male offspring dysregulation of biochemical parameters and of the insulin pathway in the hypothalamus was observed, evidencing a sex-specific mechanism.Item Efeito neuroprotetor e antiepiléptico da infusão intracerebroventricular crônica de angiotensina-(1-7) em ratos com epilepsia do lobo temporal(Universidade Federal de Goiás, 2020-03-02) Gomes, Karina Pereira; Castro, Carlos Henrique de; http://lattes.cnpq.br/6354834854727314; Colugnati, Diego Basile; http://lattes.cnpq.br/3875833705952056; Colugnati, Diego Basile; Torres, Bruno Benetti Junta; Pedrino, Gustavo Rodrigues; Blanch, Graziela Torres; Menegatti, RicardoEpilepsy is a severe neurological disorder characterized by permanent predisposition of the brain to generate spontaneous and recurrent seizures. Temporal Lobe Epilepsy (TLE) is the most frequent type of epilepsy in adults and is often refractory to pharmacological treatments available. Recently, several neuropeptides and their receptors have been suggested as promising therapeutic targets for the treatment of epilepsy, including neuropeptides and receptors of the Renin Angiotensin System (RAS). The aim of this study was to evaluate the effects of chronic treatment with Angiotensin-(1-7) (Ang-(1-7)) in male Wistar rats with TLE. The Pilocarpine (PILO) model was used for TLE induction and after the first spontaneous seizure, the animals were submitted to stereotactic surgery for implant of a guide cannula attached to an osmotic minipump. Rats of control group (CT) underwent the same procedures as the rats of groups with epilepsy, but were resistant to PILO model, not developing ELT. Intracerebroventricular infusion of Ang-(1-7) (200ng/kg/h) or sterile saline solution (NaCl 0.9%) was performed for 30 days and body weight, feed intake and spontaneous seizures were evaluated. At the end of the treatment, behavioral tests were performed in Elevated Plus Maze (EPM) and Open Field (OF) for evaluation of anxiety-like behavior and locomotor/exploratory activity. In the next day after behavioral tests, the animals were euthanized and the hippocampus were dissected, stored and later processed for protein analysis using Western Blot technique. For RAS components investigation, protein levels of Angiotensin Converting Enzyme 2 (ACE2) and Neutral Endopeptidase (NEP), as well as AT1, AT2 and Mas receptors were evaluated. In addition, hippocampal levels of Interleukin-6 (IL-6), Superoxide Dismutase (SOD), Catalase (CAT), B-Cell Lymphoma 2 (Bcl-2) and the phosphorylation of Mammalian Target of Rapamycin (mTOR) were quantified. Chronic intracerebroventricular treatment with Ang-(1-7) significantly reduced the frequency of spontaneous epileptic seizures, mainly at light photoperiod of the light-dark cycle, without changing its duration. Saline treated TLE rats (EP) showed significantly lower body weight gain than control rats without epilepsy (CT), which was attenuated in the last two weeks by Ang- (1-7) treatment (EP + Ang-(1-7)). However, the feed intake did not differ among evaluated groups. The time spent in EPM open arms by EP group was significantly higher than CT group, an effect attenuated by Ang-(1-7) treatment. In addition, the total number of entries in EPM arms was higher in EP group, which was reversed in EP + Ang-(1-7) group. The number of crossings in OF was significantly higher in EP group, which was reversed by Ang-(1-7) treatment. However, the immobility time and the numbers of rearing, also increased in the EP group, were only attenuated by Ang-(1- 7) treatment. Our results, revealed that Ang-(1-7) positively regulated the antioxidant protein CAT, the anti-apoptotic protein Bcl-2 and also increased the phosphorylation of anti-apoptotic protein mTOR, all reduced by epilepsy in hippocampus. In addition, the upregulation of AT1 receptor induced by TLE in hippocampus was also attenuated by treatment with Ang-(1-7). The protein levels of ACE2, NEP and SOD enzymes, as well as AT2 receptor and IL-6 were not statistically different between the groups. Mas receptor was downregulated in hippocampus of EP + Ang-(1-7) group. Our data suggest that Ang-(1-7) has direct modifying effects on chronic TLE by reducing oxidative stress and hippocampal neuronal death. From these results we can conclude that chronic intracerebroventricular treatment with Ang-(1-7) in rats with PILO induced- TLE reduces hippocampal neuronal damage and the frequency of spontaneous seizures, attenuating the damage in body weight gain and behavioral abnormalities. This study provides evidence that Ang-(1-7) and its receptor are promising targets for the treatment of TLE and its comorbidities.Item Efeitos celulares e comportamentais de peptídeos bioativos de baixo peso molecular extraídos de Phaseolus vulgaris(Universidade Federal de Goiás, 2020-11-18) Graziani, Daniel; Fernandes, Kátia Flávia; http://lattes.cnpq.br/9737543228759171; Custódio, Carlos Henrique Xavier; http://lattes.cnpq.br/0207928273284808; Custódio, Carlos Henrique Xavier; Cruz, Vanessa de Sousa; Araújo, Eugênio Gonçalves de; Fajemiroye, James Oluwagbamigbe; Silva, Elder Sales daINTRODUCTION: Approximately 3.000 tons of beans are not used in human consumption due to their hardening, which makes them a product of low commercial value. Several studies with bioactive peptides derived from beans have therapeutic potential to treat various diseases. OBJECTIVES: to evaluate the effects of a low molecular weight bean extract (Phaseolus vulgaris) hardened on: i) cytotoxic and cytoprotective effects on endothelial cells; ii) production of reactive oxygen species (ROS) and nitric oxide (NO) in endothelial cells; iii) oxidonitrergic-dependent vasodilator effects; iv) anxiety and depression behavior in rats; v) the antioxidant effect on rat brain. METHODS: The extract was composed of a peptide fraction less than 3 KDa (PV3) of the hardened bean residue. The PV3 sequences were corrected by mass spectrometry coupled to liquid chromatography and were analyzed with computational tools. Human umbilical vein endothelial cells (HUVEC) were treated with PV3 in the following procedures: 10 μg/ml, 20 μg/ml, 30 μg/ml and 250 μg/ml. Cellular oxidative stress was caused by 3% H2O2. Cytotoxicity and cytoprotective effects were obtained by the MTT assay, while ROS and NO were quantified by fluorescent probes (DHE and DAF-FM) by Confocal Microscopy. The vasodilator effects of NO3 and endothelium-dependent PV3 were obtained in the supplied aortic rings. The behavioral effects of acute and chronic PV3 treatments were adopted by three protocols: i) elevated plus maze test (EPM) to assess the effect of the anxiolytic type. ii) open field (OF) to assess locomotion and exploration; iii) forced swimming (NS) to test the behavior of the depressive type. The involvement of catecholaminergic pathways in the effects evoked by PV3 was tested using the enzyme tyrosine hydroxylase inhibitor, AMPT (200mg / kg). RESULTS: 35 peptides with an average mass of 1.14 KDa were identified. There was no cell death from treatment with PV3 10μg/mL and 20 μg/mL. PV3 30μg/mL increased cell viability, whereas cytotoxicity was observed only with PV3 250 μg/mL. Only PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased the release of NO without causing cell death. It was also able to reduce the relative production of cell ROS induced by H2O2. The vasodilator effects of PV3 were based on the release of NO dependent on the endothelium. In the EPM test, the acute injection of PV3 (50μg / kg) increased the frequency and the time spent in the open arms, suggesting an anxiolytic effect. In the OF test, PV3 (50μg / kg) increased the frequency of crossings and immobility time, showing that PV3 does not impair locomotion, which corroborates the anxiolytic effect found in the ECL. In the FS test, PV3 (50μg / kg) reduced the immobility time, suggesting an effect similar to the antidepressant. The anxiolytic-type effect found after acute injections was absent in chronic treatment with PV3 (50μg/kg). AMPT was able to reverse the effect of the anxiolytic type and the antidepressant type evoked by acute PV3 (50μg/kg). CONCLUSION: PV3 has low cytotoxicity, the ability to reduce ROS and increase NO in the endothelium, in addition to promoting anxiolytic and antidepressant effects with catecholaminergic involvement.Item Efeito neuroprotetor da sarcosina em modelo animal de isquemia cerebral focal(Universidade Federal de Goiás, 2022-11-04) Marques, Bruno Lemes; Pinto, Mauro Cunha Xavier; http://lattes.cnpq.br/0868250984727943; Pinto, Mauro Cunha Xavier; Vitorino, Fernanda Giachini; Castro, Célio José deStroke is characterized by a disruption in the cerebral blood supply, leading to oxygen and glucose deprivation to the tissue. Cerebral ischemia involves enhanced glutamate release, abnormal NMDAR activation, and excitotoxicity. Glycine transporter type 1 (GlyT1) modulates glutamatergic neurotransmission through NMDA receptors, suggesting an alternative stroke therapeutic strategy. This work investigated the neuroprotective and neurorestorative potential of GlyT1 inhibition in a mouse model of focal ischemia. Swiss mice subjected to permanent middle cerebral artery occlusion (MCAO) were randomized to receive three different doses of Sarcosine (125, 250, and 500 mg/kg) or vehicle before or after ischemia. Pretreatment with 250 and 500 mg/kg of Sarcosine led to neuroprotection against stroke, as demonstrated by reduction of infarct area and neurological deficits. Moreover, GluN2A/CaMKIV/CREB and BDNF/TrKB/Akt/mTOR pathways were enhanced by sarcosine. The sarcosine neuroprotection is also related to GluN2B subunit downregulation and a decrease in CaMKIIα phosphorylation. Additionally, we observed that post-stroke treatment with higher doses of Sarcosine improves the neurorepair process, which is evidenced by the marked reduction of the infarct area and motor deficits. Therefore, sarcosine pretreatment induced neuroprotection through the BDNF/TRKB and CaMKIV/CREB pathways, both processes trigged by NMDAR activity.Item Avaliação dos efeitos ambientais da vegetação urbana sobre a qualidade de vida em Goiânia(Universidade Federal de Goiás, 2001-08-31) Martins Júnior, Osmar Pires; Romão, Patrícia de Araújo; http://lattes.cnpq.br/0188184635964559; Brandão, Divino; http://lattes.cnpq.br/1614480825290154; Brandão, Divino; Casseti, Valter; Castro, Tomás de Aquino Portes eThis project is a study of the eco-system of the city of Goiânia (Goiás). Goiânia’s first Directive Plan was created according to the Garden City of Howard, one of the most important urban conceptions in the word. Using this plan and its urbanist evolution, one has tried to make a preliminary identification of the social agents, which produced the urban spaces of Goiânia. A classification and qualification of the open spaces and “green areas” of the city has been made. The quantity of vegetation (m².inhab.-1 ) and its quality (typing), when distributed adequately is important to the preservation of the urban eco-system, having important environmental effects on the quality of life of the population. The “green area” rate (Índice de Área Verde – IAV) is, therefore, one of the indicators of urban development. Changes in the open spaces, as indicators of IAV, in relation with changes in demographic density, indicative of human presence in the environment, have allowed the prevision of the amount of “green area” per urban inhabitant. The IAV calculated for Goiânia is 100,25 m². inhab.-1 , having suffered a per capita reduction of 17,68% in relation to the IAV established in the original city plan in 1938. In the next fifteen years it is possible forecast that the IAV will be 54,4% smaller than it is at the moment, decreasing to 45,71 m².inhab.-1 , if the privatization policy regarding publics spaces should persist. The “green areas” are unalienable and imprescriptible public property destined for common use. In spite of this, these areas have been decreasing over the years. The degree of deterioration of public assets has been calculated and constitutes a factor in the decay in urban life quality. The adoption of a programme to register and monitor the environmental assets of Goiânia is suggested, among other measures.Item Atividade tóxica e genotóxica induzida por combinações de antirretrovirais em Drosophila melanogaster e camundongos Mus musculus(Universidade Federal de Goiás, 2016-11-07) Moraes Filho, Aroldo Vieira de; Lee, Chen Chen; http://lattes.cnpq.br/4621907105842007; Lee, Chen Chen; Spanó, Mário Antônio; Pires, Débora de Jesus; Morais, Simone Maria Teixeira de Sabóia; Silva, Carolina Ribeiro eAs result of several mutations of HIV and failures resulting in treatments with only one medicine, it became critical to use a combination of two or more antiretrovirals in AIDS treatment protocols. So, the investigations that were carried out in this research were concentrated in the evaluation of the effects related with cytotoxicity and genotoxicity of Efavirenz (EFV) e Tenofovir (TDF) as isolated and in combination with Combivir® (AZT+3TC) and Lamivudina (3TC). For this, three test systems were used: (i) the Comet assay in Drosophila and mouse bone marrow in order to determine the genotoxic effects of the drugs tested by the DNA strand breaks; (ii) the Somatic Mutation and Recombination Test (SMART) in Drosophila melanogaster, that evaluates the toxic, mutagenic and recombinogenic activity of the compounds and (iii) the Micronucleus Test (MN) in mouse bone marrow, that detects aneugenic and clastogenic effects of agents. The results demonstrated that EFV was toxic at high concentrations and did not show induction of mutagenic and/or recombinogenic events. Inversely, Combivir and Combivir+EFV showed no lethal dose 70 (LD 70) in the concentrations used for genotoxic analysis, but induced mutagenic and/or recombinogenic effects in all tested concentrations, with prevalence of recombinogenic events. The antiretrovirals TDF, 3TC and TDF + 3TC were not toxic, but were gentoxic in all tested concentrations, with a prevalence of recombinogenicity. All of the isolated and combined compounds were positive in the Comet-assay with D. melanogaster. However, the two combinations were negative in the Comet-assay with mouse bone marrow. Combivir+EFV induced micronuclei (MN) in 24 and 48 hours. TDF+3TC induced MN only in 24 hours. Based on these results, we expect to expand the knowledge about the toxic and genotoxic activities of these combinations and to provide support for the development of new studies in AIDS treatment protocols.Item Efeito tipo-antidepressivo do derivado piperazínico 2,6-di-terc-butil-4-((4-(2-hidroxietil)piperazina-1-il)metil)fenol (LQFM212) e possíveis mecanismos de ação envolvidos(Universidade Federal de Goiás, 2023-02-27) Moreira, Lorrane Kelle da Silva; Costa, Elson Alves; http://lattes.cnpq.br/2607893423583912; Costa, Elson Alves; Fajemiroye, James Oluwagbamigbe; Ghedini, Paulo César; Rocha, Fábio Fagundes da; Carvalho, Pablinny Moreira Galdino deDepressive disorders affect individuals worldwide and may also be associated with other mental disorders such as anxiety disorders. Despite advances to improve understanding of the neurobiology of depressive disorders, no single established mechanism per se can explain all facets of these disorders and the available drugs often show therapeutic delay for clinical effectiveness. A plethora of results show the effects of piperazine derivatives on the central nervous system and are indicators of its therapeutic potential for treating mental disorders. Previously, it was shown that the piperazine derivative 2,6-di-tert-butyl-4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)phenol (LQFM212) has anxiolytic-like activity which involves serotonergic pathway, nicotinic receptors and BZD-site of GABAA receptor, without cognitive impairments. In this regard, since the same compound can have anxiolytic as well as antidepressant effects, the aim of this research was to evaluate the possible antidepressant-like activity of the compound LQFM212. Swiss albino male mice orally treated with LQFM212 (54 μmol/kg) showed behavioral effects related to antidepressant-like activity, in the forced swimming test (FST), after treatment with a single dose or with repeated doses for 15 consecutive days. Pretreatment with WAY-100635 (0.7 μmol/kg), p-chlorophenylalanine (500 μmol/kg), prazosin (2.6 μmol/kg), SCH-23390 (15 μg/kg), sulpiride (146 μmol/kg) ou α-methyl-p-tyrosine (512 μmol/kg) reversed the antidepressant-like effect of LQFM212 in the FST. Furthermore, repeated treatment with LQFM212 increased hippocampal brain-derived neurotrophic factor (BDNF) levels. Regarding the monitoring of body weight and the evaluation of possible biochemical changes the treatment with repeated doses with LQFM212 (54 μmol/kg) did not change: animals' body weight, liver glutathione (GSH) levels, and serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea and creatinine. A possible action of the compound LQFM212 on inflammatory parameters, in mice, was evaluated by systemic inflammation by the lipopolysaccharide (LPS)-induced neuroinflammation model and by local inflammation by the carrageenan- or LPS-induced pleurisy model. In the LPS-induced neuroinflammation model, oral treatment with LQFM212 (54 μmol/kg) reversed the anxiety-like and depression-like behaviors, in the open field, forced swimming and tail suspension tests, the increase of pro-inflammatory cytokines (TNF-α and IL-1β) and the decrease of anti-inflammatory cytokines (IL-4 and IL-10), into animals’ serum, caused by intraperitoneal administration of LPS (1 mg/kg). In this same model, treatment with LQFM212 (54 μmol/kg) also attenuated oxidative stress-related changes, demonstrated by reduced nitrite levels and myeloperoxidase (MPO) activity, and increased glutathione levels in the prefrontal cortex and hippocampus, and also reduced cholinesterase activity in the whole brain, of animals that received intraperitoneal administration of LPS (1 mg/kg). On the other hand, oral treatment with LQFM212 (54 μmol/kg) failed to reduce the increase in cell migration and pro-inflammatory cytokines (TNF-α and IL-1β) in pleural exudate caused by intrapleural administration of 1% carrageenan or LPS (250 ng/mL) in the pleurisy model. In addition to the reduction in MPO activity seen in the LPS-induced neuroinflammation model, treatment with LQFM212 (54 μmol/kg) also reduced the activity of this enzyme in the pleural exudate of animals subjected to the carrageenan- or LPS-induced pleurisy model. Taken together, the results showed that treatment with LQFM212 promotes behavioral changes suggestive of antidepressant-like activity in mice, which probably involve the monoaminergic pathways, in addition to increased hippocampal levels of BDNF, suggesting changes in synaptic neuroplasticity possibly as a mechanism underlying the antidepressant-like effect of the compound. The effects found in the LPS-induced neuroinflammation model did not seem to be secondary to a peripheral anti-inflammatory action of LQFM212, since this compound failed to reduce the changes caused by carrageenan or LPS in the pleurisy model. On the other hand, treatment with LQFM212 reduced MPO enzyme activity in pleural exudate, prefrontal cortex, and hippocampus, and increased per se GSH levels in both brain regions mentioned above, thus suggesting a possible antioxidant activity in vivo that may contribute to the effects observed in the neuroinflammation and pleurisy model.Item Contribuição das neurotransmissões excitatórias nos núcleos pré-óptico mediano e paraventricular do hipotálamo na modulação da atividade nervosa simpática renal e esplâncnica(Universidade Federal de Goiás, 2018-09-20) Mourão, Aline Andrade; Toney, Glenn M.; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341; Pedrino, Gustavo Rodrigues; Cavasin, Gláucia Maria; Mendes, Elizabeth Pereira; Rosa, Daniel Alves; Rebelo, Ana Cristina SilvaThe increase of sympathetic nervous activity (SNA) stands out as the main aggravating factor for diseases related to the cardiovascular system, as well arterial hypertension (AH). In fact, several research groups have directed their studies to the understanding of how alterations of regions and nuclei located in the central nervous system (CNS) could result in sympathetic hyperactivity and, consequently, the development and maintenance of AH. Cardiovascular control regions such as the median preoptic nucleus (MnPO) and the paraventricular nucleus (PVN) of the hypothalamus have been the focus of important investigations on the involvement of the CNS in the physiopathology of AH. However, signaling pathways that can modulate the neurons of these nuclei still need to be clarified. Therefore, the first part of this study aimed to investigate the involvement of angiotensinergic and glutamatergic neurotransmissions in the MnPO on the sympathetic activity and blood pressure increases observed in hypertensive rats. Spontaneously hypertensive rats (SHR) and rats submitted previously to the Goldblatt protocol (two kidneys; one clip; 2K1C) were used. Rats of both groups (250 to 350g, n=6) were anesthetized with urethane (1.2g/kg,i.v.) and instrumented to record mean arterial pressure (MAP), heart rate (HR) and renal sympathetic nerve activity (RSNA). Nanoinjection (100nl) of saline (NaCl, 150 mM), Losartan (AT1 receptor antagonist; 10mM) and kynurenic acid (glutamate receptor antagonist; 50mM) into the MnPO were performed. In 2K1C rats, glutamatergic blockade promoted decreases in MAP and RSNA (-19.1 ± 0.9 mmHg, -21.6 ± 2.8 %, p< 0.05) when compared to saline (-0.4 ± 0.6 mmHg, 0.2 ± 0.7 %, p < 0.05). Angiotensinergic inhibition also reduced these parameters (-11.5 ± 1.2 mmHg, -10.5 ± 1.0 %, p < 0.05) in 2K1C. In SHR, Kynurenic acid nanoinjections produced hypotension and sympathoinhibition (-21.0 ± 2.5 mmHg, -24.7 ± 2.4 %, p < 0.05), as well Losartan nanoinjections (-9.7 ± 1.2 mmHg; p < 0.05) and RSNA (-12.0 ± 2.4 %, p < 0.05). These findings support the conclusion that a tonic excitatory neurotransmission exerted by angiotensin II, and mostly by glutamate in the MnPO are involved in the elevation of blood pressure and RSNA observed in models of genetic hypertension or secondary to renal artery stenosis. In the second part of this study, we tested the hypothesis that the pro-inflammatory cytokine (PIC) tumor necrosis factor alpha (TNFα) could acutely activate the PVN neurons to promote SNA increased. For that purpose, Sprague-Dawley rats (350-550g) were anesthetized with a mixture of α-chloralose (80 mg / kg) and urethane (800 mg / kg) and instrumented for MAP, HR, splanchnic SNA (SSNA) recordings and unilateral nanoinjections in PVN. The animals were submitted to series of experiments with the following pretreatments in the PVN: vehicle (PBS, 0.5 nmol / 50 nl, n = 5), NBQX (AMPA receptor antagonist; 5.2 nmol / 50 nl n = 5) and AP5 (NMDA receptor antagonist; 24 nmol / 50 nl, n = 5). After each pretreatment, nanoinjection of TNFα (0.6 pmol / 50 nl) was performed and cardiovascular and sympathetic variables, evaluated for 60 min. TNFα nanoinjections in the PVN promoted progressive ramp-like increase of SSNA (55 ± 5.5 %) when pretreated with vehicle (3 ± 3 %, p < 0.05). However, no changes were observed in MAP (95 ± 8 mmHg vs. 94.5 ± 6.2 mmHg) and HR (400.2 bpm ± 13.5 vs. 377 ± 9 bpm) in this group. On the other hand, the blockade of ionotropic glutamate receptors AMPAR and NMDAR promoted reduction in the sympathoexcitatory response (24 ± 5% and 32 ± 6.5%, respectively) induced by the TNFα in the PVN. No changes in cardiovascular parameters were observed in these groups (NBQX: 93 ± 9 mmHg and 429 ± 10 bpm, AP5: 104 ± 3 mmHg and 439 ± 29 bpm). The following experiments sought to investigate the role of NMDA and AMPA in the maintenance of splanchnic sympathoexcitation caused by TNFα nanoinjections in the PVN. Thus, vehicle nanoinjections (PBS, 0.5 nmol / 50 nl, n = 3), NBQX (5.2 nmol / 50 nl, n = 5) and AP5 (24 nmol / 50 nl, n = 4) after 60 min of the TNFα nanoinjection in the PVN. The evaluated parameters were then recorded for additional 60 min. Post treatments did not promote significant alterations in any of the evaluated variables, suggesting that these receptors are not important in maintaining the sympathoexcitation triggered by the administration of TNFα in PVN. Finally, we evaluated whether the PVN inhibition could influence the maintenance of the sympathoexcitation induced by the TNFα. Thus, unilateral nanoinjections of the GABAA receptor agonist, muscimol (1 nmol / 50 nl, n = 5) were performed after 60 min of the TNFα nanoinjections in the PVN. Activation of GABAA receptors in PVN reversed TNFα-induced splanchnic sympathoexcitation from 173 ± 11% to 106 ± 5%, (p < 0.05). The results found in the present study indicate that the development of SSNA increase promoted by TNFα in the PVN is dependent of the ionotropic glutamate receptors AMPAR and NMDAR in this nucleus. Together these findings indicate the importance of glutamatergic neurotransmission in PVN for the generation of the sympathoexcitatory response triggered by neuroinflammation induced by (TNFα) in this nucleus. the MnPO are involved in the elevation of blood pressure and RSNA observed in models of genetic hypertension or secondary to renal artery stenosis. In the second part of this study, we tested the hypothesis that the pro-inflammatory cytokine (PIC) tumor necrosis factor alpha (TNFα) could acutely activate the PVN neurons to promote SNA increased. For that purpose, Sprague-Dawley rats (350-550g) were anesthetized with a mixture of α-chloralose (80 mg / kg) and urethane (800 mg / kg) and instrumented for MAP, HR, splanchnic SNA (SSNA) recordings and unilateral nanoinjections in PVN. The animals were submitted to series of experiments with the following pretreatments in the PVN: vehicle (PBS, 0.5 nmol / 50 nl, n = 5), NBQX (AMPA receptor antagonist; 5.2 nmol / 50 nl n = 5) and AP5 (NMDA receptor antagonist; 24 nmol / 50 nl, n = 5). After each pretreatment, nanoinjection of TNFα (0.6 pmol / 50 nl) was performed and cardiovascular and sympathetic variables, evaluated for 60 min. TNFα nanoinjections in the PVN promoted progressive ramp-like increase of SSNA (55 ± 5.5 %) when pretreated with vehicle (3 ± 3 %, p < 0.05). However, no changes were observed in MAP (95 ± 8 mmHg vs. 94.5 ± 6.2 mmHg) and HR (400.2 bpm ± 13.5 vs. 377 ± 9 bpm) in this group. On the other hand, the blockade of ionotropic glutamate receptors AMPAR and NMDAR promoted reduction in the sympathoexcitatory response (24 ± 5% and 32 ± 6.5%, respectively) induced by the TNFα in the PVN. No changes in cardiovascular parameters were observed in these groups (NBQX: 93 ± 9 mmHg and 429 ± 10 bpm, AP5: 104 ± 3 mmHg and 439 ± 29 bpm). The following experiments sought to investigate the role of NMDA and AMPA in the maintenance of splanchnic sympathoexcitation caused by TNFα nanoinjections in the PVN. Thus, vehicle nanoinjections (PBS, 0.5 nmol / 50 nl, n = 3), NBQX (5.2 nmol / 50 nl, n = 5) and AP5 (24 nmol / 50 nl, n = 4) after 60 min of the TNFα nanoinjection in the PVN. The evaluated parameters were then recorded for additional 60 min. Post treatments did not promote significant alterations in any of the evaluated variables, suggesting that these receptors are not important in maintaining the sympathoexcitation triggered by the administration of TNFα in PVN. Finally, we evaluated whether the PVN inhibition could influence the maintenance of the sympathoexcitation induced by the TNFα. Thus, unilateral nanoinjections of the GABAA receptor agonist, muscimol (1 nmol / 50 nl, n = 5) were performed after 60 min of the TNFα nanoinjections in the PVN. Activation of GABAA receptors in PVN reversed TNFα-induced splanchnic sympathoexcitation from 173 ± 11% to 106 ± 5%, (p < 0.05). The results found in the present study indicate that the development of SSNA increase promoted by TNFα in the PVN is dependent of the ionotropic glutamate receptors AMPAR and NMDAR in this nucleus. Together these findings indicate the importance of glutamatergic neurotransmission in PVN for the generation of the sympathoexcitatory response triggered by neuroinflammation induced by (TNFα) in this nucleus.Item Análise da dinâmica do glicoproteoma de trichoderma asperelloides durante o micoparasitismo(Universidade Federal de Goiás, 2022-01-21) Naoum, Stéphanie; Monteiro, Valdirene Neves; http://lattes.cnpq.br/8264822485508916; Ulhoa, Cirano José; http://lattes.cnpq.br/8368469162867277; Ulhoa, Cirano José; Bailão, Alexandre Melo; Georg, Raphaela de Castro; Paula, Renato Graciano deGlycosylation is a post-translational modification that occurs in most cells and is an important mechanism for several cellular processes such as protein secretion, cell signaling, protein translocation and stability, maintenance of cell structure and receptor-ligand interactions. In fungi there is a set of enzymes specialized in glycosylation of proteins, exerting functions related to the structure of the cell wall and the cell as a whole, assisting in integrity, growth, differentiation and signaling. Fungi of the genus Trichoderma are known for their ability to biocontrol through mycoparasitism mechanisms involving the production of cell wall degrading hydrolytic enzymes. Therefore, the objective of this work was to identify the N-glycosylated proteins produced by T. asperelloides (TR356) during mycoparasitism. The Concanavalin A (ConA) affinity chromatography technique was used to enrich the samples and select N-glycated glycoproteins with oligomannosidic structure. In the interaction between the fungi, the contact condition showed a difference in the protein content when compared to the control samples. The specific activities of the enzymes β-1,3-exoglycanase, β-1,3-endoglycanase, chitinase, N-acetyl-glucosaminidase, β-glucosidase, acid phosphatase, α-mannosidase, α-arabinofuranosidase and demonstrated a significant increase in activities in conditions before contact and contact. Several proteins were identified in the samples using the mass spectrometry technique. A total of 253 proteins were identified in the control sample. In samples referring to before contact and contact between T. asperelloides (TR 356) and S. sclerotiorum the number of identified proteins was higher, 582 and 524 proteins, respectively. We can infer that the presence of the pathogen S. sclerotiorum in the same environment as T. asperelloides stimulates the production of specific proteins for this situation, necessary for mycoparasitism. Glycoproteins with different amounts of N-glycosylation sites involved in mycoparasitism were identified, and the number of glycoproteins and N-glycosylation sites increased in pre-contact and in-contact situations. Most secreted proteins are involved in carbohydrate metabolism and transport, cell signaling, and post-translational modifications and folding. The identified proteins of the intracellular environment are involved in post-translational modification and protein folding, in carbohydrate metabolism and transport, and in cellular metabolism in general. Finally, we observed that a significant number of identified proteins still do not have a defined function, which can be considered an important source of new studies and new knowledge in relation to mycoparasitism.Item Contribuição da neurotransmissão glutamatérgica na região rostroventrolateral do bulbo (RVLM) para as respostas cardiovasculares e autonômicas induzidas pelo fator de necrose tumoral alfa (TNF-α) no núcleo paraventricular do hipotálamo (PVN)(Universidade Federal de Goiás, 2022-10-31) Naves, Lara Marques; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341; Pedrino, Gustavo Rodrigues; Custódio, Carlos Henrique Xavier; Fajemiroye, James Oluwagbamigbe; Mourão, Aline Andrade; Oliveira, André Henrique Freiria deNeurogenic hypertension is characterized by a chronic elevation of blood pressure (BP) associated with exacerbation of sympathetic nerve activity (SNA). In this sense, the neuroinflammation, marked by the presence of pro-inflammatory cytokines (PIC) in the central nervous system (CNS), can be related to increased sympathetic drive and arterial hypertension (AH) development. Furthermore, the presence of tumor necrosis factor alpha (TNF-α) in sympathetic premotor neurons that compose the rostral ventrolateral medulla (RVLM) and hypothalamic paraventricular nucleus (PVN) is associated with hypertensive phenotype. However, the pathways and mechanisms by which TNF-α act in the CNS remain under investigation. Thus, the present study investigated the cardiovascular and autonomic effects promoted by TNF-α administration in the PVN and the participation of glutamatergic neurotransmission and N-methyl-D-aspartate (NMDA) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in the RVLM in these responses. For this, Wistar and spontaneously hypertensive rats (SHR) (270 - 300g) were anesthetized with urethane (400 mg/mL, intravenous - i.v.) associated with α-chloralose (40 mg/mL, i.v.) and instrumented to mean arterial pressure (MAP), heart rate (HR) and splanchnic sympathetic nervous activity (SSNA) recordings. The animals were organized into five groups and subjected to unilateral nanoinjections (50 nL) in the RVLM as follows: I. Wistar subjected to vehicle nanoinjections (Ringer's solution, normotensive SHAM group, n=5); II. SHR subjected to vehicle nanoinjections (Ringer's solution, hypertensive SHAM group, n=7); III. SHR subjected to kynurenic acid nanoinjections (KYN, 50 mM, glutamate receptor antagonist, hypertensive GLU group, n=6); IV. SHR subjected to 2-amino-5-phosphonovaleric acid nanoinjections (AP5, 24 nmol/50 nL, NMDA receptor antagonist, hypertensive NMDA group, n=7) and V. SHR subjected to 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f] quinoxaline nanoinjections (NBQX, 5.2 nmol/50 nL, AMPA receptor antagonist, hypertensive AMPA group, n=6). Then, all groups were subjected to ipsilateral TNF-α nanoinjections (0.6 pmol/50 nL, 50 nL) in the PVN. Nanoinjections of vehicle, KYN, AP5 or NBQX in RVLM did not change baseline values of MAP, HR and SSNA. In the normotensive SHAM group, TNF-α nanoinjections into the PVN induced an ANSE increase after 50 min of TNF-α nanoinjections, without modifying the MAP and HR. In contrast, in the hypertensive SHAM group, TNF-α nanoinjections in the PVN promoted a progressive splanchnic sympathoexcitation initiated 20 min after the TNF-α nanoinjections. After 50 min of TNF-α nanoinjections, a pressor response was observed, without changing HR, in the hypertensive SHAM group. Previous inhibition of RVLM glutamatergic neurotransmission did not alter the pressor response induced by TNF-α in hypertensive animals and did not change the HR. However, abolished the splanchnic sympathoexcitation observed after TNF-α. Additionally, NMDA or AMPA receptors previous inhibition in RVLM was not able to change the TNF-α-induced late increase in MAP and the HR in hypertensive animals. However, attenuated the splanchnic sympatoexcitation generated after the TNF-α nanoinjections. These results suggest that cardiovascular and autonomic changes promoted by TNF-α in the PVN are exacerbated in hypertensive animals and that the integrity of glutamatergic neurotransmission and NMDA and AMPA receptors in the RVLM are essential for the sympathoexcitatory response induced by TNF-α in the PVN in SHR.Item Avaliação citotóxica de nanopartículas magnéticas utilizando fotohipertermia no tratamento de melanoma(Universidade Federal de Goiás, 2019-03-23) Oliveira, André Luiz Silva; Bakuzis, Andris Figueiroa; http://lattes.cnpq.br/3477269475651042; Lacerda, Elisângela de Paula Silveira; http://lattes.cnpq.br/9390789693192751; Lacerda, Elisângela de Paula Silveira; Lee, Chen Chen; Cardoso, Clever GomesJustification: Nanophototherapy has been shown a promising technique in non-invasive treatment of high selectivity with active uptake to the tumor target, showing good intracellular hyperthermic heat delivery efficiency, which promotes therapeutic effects such as tumor regression and increased expression of Heat Shock Proteins (HSPs). These proteins have also been shown to trigger systemic processes such as thermal immunoactivation and abscopal effect. It has been invested in the search for new nanoformulations that present the characteristics of biocompatibility required for nanophotohyperthermic applications. In this context, a study was conducted on the cytotoxic and antitumor profile in order to contribute to the development of new drugs, which are more efficient and safer for cancer treatment. Objectives: To evaluate cell viability, cytotoxic effect, mechanism of death after intracellular thermal treatment of nanophotohyperthermia with MALB magnetic nanoparticle (BSA+MnFe2O4) on murine melanoma tumor cells (B16-F10) and normal murine fibroblast cell line (L-929). Methodology: Cells were cultured in 6-well plates with DMEM containing 10% FBS, 1% of penicillin, in a humidified incubator at 37°C with 5% CO2. It were plated 5x105 cells per well after 3h initiated incubation with MALB for 12h or 24h at a final concentration of 914μg.mL-1 (proportion of 547 cells per ug of MALB). After the endocytosis time, the cells were washed twice in PBS and centrifuged obtaining pellets to receive nanophotohyperthermia treatment for 30min (ʎ=808nm, potency=4-6W/cm2). The heating was captured by infrared thermal camera images. After intracellular heat treatment at 42.5 or 46°C were verified the cytotoxicity, cell viability, the mechanism of death and ability to form new colonies. Statistical analysis of ANOVA was done using GraphPad Prism 5. Results: MTT assay results non-heating promoted by nanophotohyperthermia make it clear that MALB and MnFe2O4 coated with sodium citrate do not present cytotoxicity for both lineages. The MALB is selective to the melanoma tumor line. Evidence was found that MALB has active uptake to the tumor target, due to albumin mounted on the nanocarrier. It was observed that MALB [914μg.mL-1] previously incubated for 12 or 24h more laser application has led tumor cells B16-F10 to high intracellular hyperthermia temperatures range (42.5 and 46°C) resulting in 79% of death by late apoptosis (annexin V/+ and PI/+), due to thermal necrosis and only 17% of living cells. Through iron mass revealed by Ferene-S assay has been found that MALB 24h incubation was endocytosed 10.67 ± 0,5 pg Fe+/cell (B16-F10), the equivalent 3.5x higher than control (-) and still twice greater than for L-929. Conclusion: MALB showed good intracellular heat delivery efficiency in a dose-dependent manner. The nanophotohyperthermia heating tests presented good results of destruction of micro-phantom tumors for melanoma model in vitro (B16-F10) and still showed high selectivity to the tumoral lineage, due to the presence of albumin (BSA) in its nanostructure, revealing that this nanocarrier has active tumor targeted characteristic