Mestrado em Ciências Biológicas (ICB)

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    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;; Gomes, Rodrigo Mello; Mascioli, Cristina da Costa Krewer; Custódio, Carlos Henrique Xavier
    Isoproterenol 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.
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    Avaliação da atividade genotóxica e antigenotóxica de Celtis iguanaea (Jacq.) Sargent em bactérias e camundongos
    (Universidade Federal de Goiás, 2010-02-24) Borges, Flávio Fernandes Veloso; Lee, Chen Chen;
    Celtis iguanaea (Jacq.) Sargent is a member of the Cannabaceae family and popularly known as esporão de galo. Ethnobotanical surveys of cerrado native plants show that the tea leaves of C. iguanaea is used in traditional medicine for body aches, asthma, cramps, poor digestion, urinary infections, kidney disfunctions, as a stimulant or as a diuretic.The purpose of the present work was to evaluate the possible cytotoxic, genotoxic and antigenotoxic effects of the leaves aqueous extract of C. iguanaea by the prophage λ induction test (SOS Inductest) and the micronucleous test in mice bone marrow. The SOS Inductest was performed according to Moreau (1976), using lysogenic WP2s(λ) and indicator RJF013 strains derived from Escherichia coli. WP2s(λ) cultures were treated with different doses of the extract (0.5, 1, 2.5, and 5 mg) (evaluation of genotoxicity) or co-treated with a single dose of MMC (0,5 mg) and different doses of extract (assessment of antigenotoxicity). Then, the treated culture was added to the indicator strain (RJF013) and both were poured on plates in LB(1/2) medium. For the evaluation of cytotoxicity, cultures of WP2s(λ) treated with different doses of extract were diluted in M9 buffer and plated on LB. The micronucleus test was carried out according to Schmid (1975). To assess the genotoxic activity, animals were treated with Celtis iguanaea extract (100, 300 and 500 mg/kg concentrations). To evaluate the antigenotoxic activity, the same doses of extract were treated simultaneously with 4mg/Kg of MMC. The frequencies of micronucleated polychromatic erythrocytes (MNPCE) were evaluated at 24h and 48h exposure period except the negative control (24h). The cytotoxicity was assessed by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). In both tests the obtained results demonstrated anticytotoxic activity and absence of genotoxicity and cytotoxicity of the Celtis iguanaea leaves extract. The extract also showed partial antigenotoxic activity in bacteria and antigenotoxic activity in mice.
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    Planejamento e síntese de novos candidatos a protótipos de fármacos anti-crises epiléticas
    (Universidade Federal de Goiás, 2023-03-06) Rocha, André Luis Batista da; Arruda, Evilanna Lima;; Menegatti, Ricardo;; Rocha, André Luis Batista da; Menegatti, Ricardo; Colugnati, Diego Basile; Martins, Táric Ramon Marques
    Epilepsy is a disease that affects about 50 million people worldwide and the worldwide prevalence is estimated at 1% of the population, and of this global rate, about 30% of patients are refractory to pharmacological treatment. In Brazil, 150 thousand new cases are registered per year. Therefore, this study aims at planning and developing new candidates for prototype anti-epileptic seizure drugs that are safer, more effective and that present fewer adverse reactions, or that can be used as a pharmacological tool. In this situation, the work describes the synthesis of compounds analogous to stiripentol. Ring bioisosterism and hybridization strategies were employed in the planning. To obtain the compounds, a synthetic route with four steps was chosen. The intermediate compound was obtained through the aldol condensation method and the final compound was obtained by the Luche reduction method, which obtained an overall yield in the range of 21% to 23%. The synthesized compounds, with the exception of compound LQFM 311, were within the parameters established by Lipinski and Verber. The compounds obtained in the synthesis were purified by recrystallization method employing solvent mixture and the purification of compound LQFM 311 was performed by column adsorption chromatography. All synthesized compounds were characterized by hydrogen nuclear magnetic resonance (¹H NMR) and ¹³C nuclear magnetic resonance (HSQC and HMBC). This demonstrates that the chosen synthetic route was adequate, allowing the development of new drug prototypes.
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    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;; Pedrino, Gustavo Rodrigues;; Pedrino, Gustavo Rodrigues; Costa, Renata Mazaro; Ferreira, Patricia Maria
    Diseases 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.
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    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;; Georg, Raphaela de Castro; Coelho, Alexandre Siqueira Guedes; Steindorff, Andrei Stecca
    The 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.
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    Efeito neuroprotetor da inibição do transportador de glicina do tipo 1 em modelos animais de dano estriatal
    (Universidade Federal de Goiás, 2022-12-16) Ribeiro, Raul Izidoro Ribeiro; Pinto, Mauro Cunha Xavier;; Pinto, Mauro Cunha Xavier; Giacomelli, Ágatha Oliveira; Vieira, Luciene Bruno
    The striatum is a region of the brain associated with motor control, with changes in its function associated with neurodegenerative diseases such as Parkinson's and Huntington's. Although several factors are related to the neurodegeneration process in these diseases, the phenomenon of excitotoxicity and excessive stimulation of NMDA receptors (NMDAR) is a common point between them. An approach to prevent excitotoxicity is associated with the blockade of GlyT-1, which promotes the increase of glycine in the synaptic cleft, promoting a profound neuromodulation process. Based on this evidence, the objective of this work was to evaluate the effect of pretreatment with a type 1 glycine transporter inhibitor (NFPS) in animal models of Parkinson's disease (PD) and Huntington's disease (HD). Open field tests, cylinder test, rotarod and apomorphine-induced rotation tests were used to monitor motor changes in the experimental models. Western Blotting technique was performed to evaluate protein expression upon treatment with NFPS. The results suggest a neuroprotective effect, by preventing a reduction in movement, reducing striatal damage, and increasing the expression of proteins related to cell survival. Taken together, our data suggest that GlyT-1 inhibition has neuroprotective potential for striatal lesions associated with HD and PD.
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    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;; Gomes, Rodrigo Mello; Miranda, Rosiane Aparecida; Costa, Renata Mazaro e
    Among 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.
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    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;; Pinto, Mauro Cunha Xavier; Vitorino, Fernanda Giachini; Castro, Célio José de
    Stroke 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.
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    Estudo morfofisiológico dos efeitos do flavonoide crisina sobre o desenvolvimento pré-puberal da próstata de gerbilos (meriones unguiculatus)
    (Universidade Federal de Goiás, 2017-07-24) Ribeiro, Naiara Cristina de Souza; Perez, Ana Paula da Silva;; Santos, Fernanda Cristina Alcantara dos;; Santos, Fernanda Cristina Alcantara dos; Georg, Raphaela de Castro; Oliveira, Sérgio Marcelino de
    The prostate is a gland of the male reproductive system accessory, but not exclusively of males, and can be found in females of various mammals. Changes in hormone metabolism mediated by exogenous substances can interfere with the activity of this gland. An example is chrysin (5,7-dihydroflavone), a natural compound found in plants. This substance modulates the action of sex hormones, interfering with the synthesis and metabolism of steroid hormones. The objective of this study was to evaluate the effects of exposure to chrysin on the prepubertal development of the prostate of gerbils of both sexes. Thirty males and 30 females of adult gerbils were used. Each of these females was maintained with a male of the same age for mating to occur. After the cubs were born, three experimental groups were formed: 1) Control (dilution vehicle); 2) chrysin (25 mg/kg/day); 3) Positive Control (1 mg/kg/week testosterone cypionate). Males and females of all groups were treated from the 15th to the 42nd postnatal life and euthanized on the 43rd day (onset of puberty) and 90 days (onset adult). The organs were submitted to morphological, stereological and Immunohistochemical analysis. The results showed that pre-pubertal exposure to chrysin did not alter the structural parameters of the prostate of pubertal gerbils, thus increasing the frequency of androgenic (AR) and alpha estrogen (ERα) receptors. Testosterone promoted an increase in the volume and secretory activity of the alveoli at this stage of development. In male and female adult animals, chrysin promoted an increase of secretory epithelium presenting highly developed prostate glands, with a high rate of cell proliferation and high secretory activity. In addition, it maintained a significant increase in the rate of cell proliferation and the frequency of AR, and ERα-positive cells. The results allowed to conclude that the pre-pubertal exposure to chrysin caused permanent changes in development, which only became noticeable in adult life. Studies indicate that pre-pubertal exposure to flavonoid chrysin causes hormonal disorders that are capable of permanently altering prostate homeostasis. Since the high consumption of these flavonoids during the developmental stages one must be more careful since chrysin permanently changes the mofophisiology of hormone-dependent organs.
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    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;; Pinto, Mauro Cunha Xavier; Menegatti, Ricardo; Leite, Jacqueline Alves
    Schizophrenia 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.
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    Avaliação da atividade genotóxica, citotóxica e histopatológica de imunossupressores em diversos níveis celulares e organismos
    (Universidade Federal de Goiás, 2019-08-29) Oliveira, Laís Camargo de; Carneiro, Cristiene Costa;; Lee, Chen Chen;; Lee, Chen Chen; Silva, Carolina Ribeiro e; Reis, Paulo Roberto de Melo
    Prednisone (PD) and azathioprine (AZA) are widely used in the immunossupressive therapy of transplanted and autoimmune disease patients. The chronic use of immunossupressants has been correlated with the occurrence of several side effects and also with the increase of neoplasia in those patients. Considering that genotoxic activity is associated to the increase of risk of cancer development, this study evaluated mutagenic, genotoxic and cytotoxic activities PD and AZA using in vitro and in vivo assays. The mutagenic activity was evaluated using mutagenicity Ames test in Salmonella typhimurium TA100 strain. The evaluation of genotoxicity and cytotoxicity was performed using micronucleus and comet assays in mouse bone marrow cells. Moreover, it was evaluated the toxic effects of PD and AZA in organs (liver and kidneys) of mice. In the mutagenic Ames test PD didn’t showed significant difference compared to the negative control in all tested doses (p > 0,05). In contrast, AZA exhibited significant increase compared to negative control in the two higher doses (20 and 40 μg/plate) (p < 0,05). In the micronucleus test, PD showed genotoxic activity in all tested doses and times and was cytotoxic in two of the tested doses (1,0 mg/kg in 48 hours and 0,5 mg/kg in 120 hours). In the comet assay, PD showed high porcentage of DNA damage in all tested doses and times. Furthermore, the 120 hours treatment presented the highest genotoxic activity when compared to the other PD tested groups (p < 0,05). In the same assay, AZA showed significant increase in all doses and times of treatment when compared with negative control (p < 0,05). In the histopathological analyzes, it was observed a moderate to accentuated congestion in mice liver treated with PD and a discrete hyperemia in mice liver when treated with AZA. Therefore, PD did not showed mutagenic activity in Ames test, however it was genotoxic and cytotoxic in the in vivo assays, and presented moderated toxic effect in the histopatological analyses. AZA exhibited mutagenic and genotoxic activity in several genotoxicological assays performed, however it did not cause relevant toxicity in mice tissues.
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    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;; Ghedini, Paulo César; Duarte, Djane Braz; Munhoz, Carolina Demarchi
    Chronic 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.
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    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;; Lacerda, Elisângela de Paula Silveira;; Lacerda, Elisângela de Paula Silveira; Lee, Chen Chen; Cardoso, Clever Gomes
    Justification: 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
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    Recuperação cardiovascular induzida por infusão de solução salina hipertônica em ratos hemorrágicos: participação dos receptores adrenérgicos no órgão subfornical
    (Universidade Federal de Goiás, 2021-01-28) Silva, Amanda Barbosa Coelho da; Fajemiroye, James Oluwagbamigme;; Pedrino, Gustavo Rodrigues;; Pedrino, Gustavo Rodrigues; Fajemiroye, James Oluwagbamigbe; Mourão, Aline Andrade; Custodio, Carlos Henrique Xavier
    Previous studies have shown that the immediate restoration of cardiovascular parameters, such as blood pressure and cardiac output, by hypertonic saline solution (HSS) is useful in the treatment of hypotensive hemorrhage (HH). The Subfornical Organ (SFO) receives projections of regions involved in osmotic and cardiovascular control, and the integration of these neuronal regulatory pathways by SFO is assumed to play a role in SSH-induced cardiovascular recovery in hemorrhagic rats. Thus, the present study assessed the role of SFO and adrenergic pathways in cardiovascular responses to HSS infusion in hemorrhagic rats. All experiments were approved by the Ethics Committee on the Use of Animals at the Federal University of Goiás (CEUA-UFG; protocol nº 034/12). Wistar rats (270–300g) were anesthetized with halothane (2% in 98% O2; Tanohalo; Cristália, Itapira, SP, Brazil) and after insertion of the venous catheter, anesthesia was maintained by urethane (1.2 g ∙ kg- 1, iv; Sigma-Aldrich, MO, USA). The animals were instrumented to record mean arterial pressure (MAP), heart rate (HR), renal blood flow (RBF) and aortic (ABF). The values of renal vascular conductance (RCV) and aortic (AVC) were calculated from the ratio between RBF or RBA and MAP, respectively. The HH was induced by withdrawing blood over 10 min until MAP reached approximately 60 mmHg, after which this MAP level was maintained for another 10 minutes by withdrawing or reinfusing blood when necessary. After 10 min of blood withdrawal, saline (NaCl; 0.15M; CONT; n = 5), muscimol (4mM; MUSC; GABAergic agonist; n = 6) or propranolol (10mM; PROP; non-selective β-adrenergic blocker; n = 6) were nanoinjected (100nL) in SFO. The sodium overload, through the infusion of HSS (NaCl 3 M; 1.8 ml ∙ kg-1 of body mass), was performed 20 min after the HH. The HH promoted hypotension (CONT: from 103.3 ± 3.5 to 62 ± 0.3 mmHg; MUSC: from 108 ± 4.4 to 61 ± 0.8 mmHg and PROP: from 98.4 ± 2, 4 to 61 ± 1.3 mmHg; 20 min after HH; p <0.05). The sodium overload promoted MAP restoration to values close to baseline in the animals that received saline nanoinjections (92 ± 3.1 mmHg; 40 min after HSS infusion; p <0.05). However, in animals that received nanoinjections of muscimol and propranolol, the HSS infusion did not restore this parameter to baseline levels (MUSC: 53.0 ± 3.8 mmHg and PROP: 59 ± 4.8 mmHg; 40 min after HSS infusion; p <0.05; compared to baseline). The changes in HR values were not significant in all the groups (CONT: from 402.9 ± 13.4 bpm to 381.0 ± 17.1; MUSC: from 408.0 ± 10.5 bpm to 375.0 ± 22.3 bpm and PROP: from 410 ± 15.6 to 362 ± 14.6 bpm; 20 min after HH). After 40 minutes of HSS infusion, there is no significant change in this parameter (CONT: 346.0 ± 19.7 bpm; MUSC: 352.0 ± 18.1 bpm and PROP: 336.9 ± 14.2 bpm). The HH promoted a significant reduction in RBF in all groups (CONT: ∆: -59.8 ± 5.3%; MUSC: ∆: -53.4 ± 14.6% and PROP: ∆: 48.7 ± 6.8%, in relation to the baseline, p <0.05; 20 minutes after HH). The HSS infusion did not restore the RBF values in the control and experimental groups (CONT: ∆: -20.8 ± 19.1%; MUSC: ∆: -64.9 ± 4.1% and PROP: ∆: -51.3 ± 11.6%, in relation to baseline, p <0.05; 40 minutes after HSS infusion). The was no significant differences in the value of RVC after HH (CONT: ∆: -33.6 ± 8%; MUSC: ∆: -23 ± 6.8% and PROP: 17: -17.4 ± 10.3%, 20 min after HH) and HSS infusion (CONT: ∆: -32.6 ± 2.9%; MUSC: ∆: -27 ± 8.0% and PROP: ∆: - 14.5 ± 15.7%; 40 minutes after HSS infusion). The HH significantly reduced the ABF in the groups (CONT: ∆: -75 ± 5.2%; MUSC: ∆: -60.1 ± 9.0 and PROP: ∆: -57 ± 5%, p <0.05; 20 min after HH), and this reduction was maintained even after 40 minutes of the HSS infusion (CONT; ∆: - 55 ± 5.8%; MUSC: ∆: -57 ± 6.9% and PROP: ∆: -60 ± 3.7% compared to baseline, 40 min after HSS infusion). No changes were observed in the AVC after HH (CONT: ∆: -46.4 ± 14.1%; MUSC: ∆: -30.3 ± 15.1% and PROP: ∆: -30.4 ± 7.4%, p <0.05; 20 min after HH) and HSS infusion (CONT: ∆: -52.2 ± 4.5%; MUSC: ∆: -11.4 ± 14.8 % and PROP: ∆: -33.6 ± 4.1%, 40 minutes after HSS infusion; in relation to baseline). These findings strengthen the hypothesis of the SFO involvement in HSS-induced cardiovascular recovery during HH, and suggest the attenuation of this recovery by pharmacological blockade of β-adrenergic neurotransmission. In this manner, the dysfunction of adrenergic neurotransmission in SFO could prevent HSS-induced cardiovascular recovery after HH.
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    Dessincronização forçada da atividade/repouso durante o período gestacional de ratas Wistar: quais as consequências sobre os parâmetros cardiometabólicos observadas na prole?
    (Universidade Federal de Goiás, 2021-04-27) Xavier, Isabela Cristina Maioni; Rosa, Daniel Alves;; Rosa, Daniel Alves; Mendes, Elizabeth Pereira; Castro, Carlos Henrique de
    The circadian system disruption has been associated with several health problems. In shifts work women there is a higher incidence of metabolic disorders and problems during the gestational period. Thus, we sought to determine whether the induction of forced internal circadian desynchronization during the pregnancy of Wistar rats would lead to reproductive changes or metabolic disorders in mothers after lactation period, as well as whether there would be cardiometabolic consequences in their adulthood offspring. Methods: Wistar rats were randomly assigned to a group: control (CTR), subjected to a light / dark (LD) 12h (T24) cycle; and desynchronized (DSC) subjected to a symmetrical LD cycle of 11h (T22) during pregnancy. The respective offspring (pCTR and pDSC) were followed for 90 days. In the mothers, the oral glucose tolerance test (oGTT) and the intraperitoneal insulin tolerance test (ipITT) were performed on the 22nd postnatal day, as well as in the puppies at 90 days. For 8 weeks, the systolic blood pressure (SBP) and heart rate (HR) in the offspring were verified through tail plethysmography. After 24 hours of the oGTT test, all animals were euthanized for tissue collection. Results: DSC rats exhibited a higher percentage of locomotor activity during the light phase and a lower percentage in the dark phase of the LD cycle than the CTR during the gestational period, without difference in total. There were no changes in body weight gain and relative intake food and water between groups. Regarding reproductive aspects, there were no significant differences between groups or in the number of pups per litter as for body weight at birth. After weaning, only DSC mothers had a lower glucose decay constant (Kitt) in the ipITT test. The pDSC showed higher weight gain in the 7th and 8th week after weaning. However, there were no significant differences in the epididymal, mesenteric, inguinal and retroperitoneal adipose tissue mass between groups. SBP and HR over 8 weeks also showed no significant differences between groups. After 90 days, the pDSC showed higher heart weight, pancreas and brown adipose tissue and a reduction in renal mass when compared to their control pairs. However, after lactation, DSC mothers showed a reduction in plasma glucose uptake, suggesting glucose intolerance or impaired insulin sensitivity. Although pDSC in adulthood showed higher gain in total body mass and changes in tissue weights that are important for the balance of cardiometabolic functions, it was not possible to detect significant changes in PAS and HR levels, nor in glycemic metabolism tests. Histological analysis of renal tissue did not show any changes, whereas cardiac analysis showed a greater deposition of interstitial and perivascular fibrosis. These results suggest that forced internal desynchronization during pregnancy did not result in negative reproductive consequences for mothers; however, it was responsible for a cardiac tissue remodeling adult pDSC.
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    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;; Brandão, Divino;; Brandão, Divino; Casseti, Valter; Castro, Tomás de Aquino Portes e
    This 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.
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    Fluorescência da clorofila a e liberação de O2 em diferentes variedades de feijão (Phaseolus vulgaris) e soja (Glycine Max(L.)Merr.) após aplicação de sulfato de amônio
    (Universidade Federal de Goiás, 2006-11-25) Silva, Kelly Pereira da; Castro, Tomás de Aquino Portes e;; Castro, Tomás de Aquino Portes e; Fageria, Nand Kumar; Didonet, Agostinho Dirceu
    The productivity of vegetables is influenced by weather, physiology and genetics associated by photosynthesis, by the division of photoassimilated, and by the fixation of N. The analysis of fluorescence of clorophyll allows the study of the characteristics related with the capability of absorption and transference of the luminous energy in the chain of electron transport. The aim of the present research was to test three varieties of kidney bean and three varieties of soy bean influenced by nitrogen in the exchanges of O2 and the fotochemical efficiency (Ef), using techniques of emission of fluorescence and O2 envolved during photosynthesis, comparing each other. The determinated variants were: minimum fluorescence (F0), maximum fluorescence (Fm), terminal fluorescence (Ft), variable fluorescence (Fv), efficiency quencher (Eq) and fotochemical efficiency (Ef). The O2 envolved during photosynthesis was avaliated. The plants were submitted to the recommended level of N and the determinations of fluorescence emission (F0, Fm, Ft, Fv, Fv/Fm) and the O2 envolved during photosynthesis were realized one day after the application of NH4+ like ammonium sulfate. Related to fluorescence there was not difference between the varieties of kidney bean and soy bean. When it was analised the application of NH4+ it was observed that there was difference in the statistic of soy bean. It was observed the O2 envolved during photosynthesis in the variety of kidney bean Pérola ( 1,50 mol de O2 m –2 s –1) was greater than the other and in the variety of soy bean Monsoy ( 1,58 mol de O2 m –2 s –1) was greater than the other. Additional index terms: O2 evolution, Phaseolus vulgaris, Glycine Max, fluorescence, fotochemical efficiency, quencher efficiency.
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    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;; Faria, Fabrícia Paula de;; Faria, Fabrícia Paula de; Lopes, Francis Júlio Fagundes; Ulhoa, Cirano José
    The 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).
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    Caracterização e avaliação de apneias obstrutivas induzidas em ratos com epilepsia e seu impacto no sono, respostas cardiovasculares e de esforço respiratório
    (Universidade Federal de Goiás, 2020-02-28) Lima Júnior, Cláudio Quintino de; Pansani, Aline Priscila;; Colugnati, Diego Basile;; Colugnati, Diego Basile; Pansani, Aline Priscila; Rosa, Daniel Alves; Costa, Renata Mazaro
    Epilepsy is one of the most common neurological diseases. Mortality rates are considerably higher in individuals with epilepsy and the most common category of death related is Sudden Unexpected Death in Epilepsy (SUDEP). Multifactorial mechanisms related to autonomic and respiratory cardiac changes underlying SUDEP. Obstructive sleep apnea (OSA) has a high incidence in patients with epilepsy and it is a risk factor for SUDEP. The aim of this study was to characterize OSA in animals with epilepsy. For this, Wistar rats (230-250g) were submitted to Pilocarpine model of epilepsy. After 30-60 days of chronic epilepsy, the animals were submitted to surgical procedures to evaluation of sleep and heart rate, respiratory effort and induction of obstructive apneas. Rats with epilepsy had altered basal sleep, with decreasing REM in 24-hour record and increased wakefulness and decreased NREM sleep in the dark phase of the cycle. Both NREM and REM sleep of these rats had a higher percentage of delta waves, and REM had lower theta waves. Rats with epilepsy had higher spontaneous central sleep apneas. Control rats had REM-sleep apneas longer than NREM-sleep apneas. This difference did not occur in rats with epilepsy. OSA did not altered sleep fraction in any group. In Epilepsy group, there was decrease in REM fraction at the first 8 h of recovery sleep of REM-apneas. Cardiovascular and respiratory effort during OSA and chemoreflex response were similar between groups. However, in epilepsy, there was a tendency to low in both respiratory effort in awake apneas and pressure response induced bychemoreflex. In epilepsy, there was higher density of NK1 in pré-Bötzinger and lower number of serotonergic neurons in Raphe Magnus and Pallidus. Therefore, rats with epilepsy had alteration on sleep-wake cycle and in sleep-delta/theta ratio. Also, the mechanisms related to end of OSA were altered, possibly by alteration in neurons of respiratory control.
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    Via descendente rostroventromedial bulbar que medeia as respostas cardiovasculares evocadas pela coluna dorsolateral da substância cinzenta periaquedutal
    (Universidade Federal de Goiás, 2020-03-10) Moraes, Gean Carlos Alves; Lima, Onésia Cristina Oliveira;; Custódio, Carlos Henrique Xavier;; Custódio, Carlos Henrique Xavier; Cruz, Kellen Rosa da; Oliveira, Patrícia Maria de
    Periaqueductal gray is a midbrain region surrounding the cerebral aqueduct that projects to areas controlling behavioral and autonomic outputs. The activity of the lateral and dorsolateral columns of PAG is required for expressing the behavioral and physiological components of defense reactions. Cardiovascular responses evoked from PAG include increases in blood pressure, positive chronotropism and regional tissue perfusion changes. However, literature is scarce on the descending pathways controlling cardiovascular responses evoked from PAG. Since Raphe Pallidus (RPa) is a medullary region comprising sympathetic premotor neurons projecting to preganglionic spinal segments connected to sympathetic supplies innervating the heart, it is worth considering the PAG-RPa path. i) to assess whether PAG projects to RPa; ii) to evaluate the amplitude of the inotropic and chronotropic responses evoked from PAG; iii) to assess whether cardiovascular responses evoked from PAG rely on RPa. Experiments were conducted in Wistar rats (300g) and were approved by CEUA - UFG (092/18). In a first set of experiments (n=3), monosynaptic retrograde tracer Retrobeads were injected into RPa, and PAG slices were analyzed. Other two groups of (n = 6 each) were anesthetized with urethane (1.2 g / kg) and chloralose (120 mg / kg) ip and underwent tracheostomy, cannulation of the femoral artery and vein, catheterization of cardiac left ventricular and craniotomy. After a cardiovascular parameters’ stabilization, one group was injected vehicle into RPa 20min before injection of the GABAA receptor antagonist, bicuculline Methiodide (BIC 40 pmol / 100nL) into the lateral / dorsolateral PAG. The other group was injected with the GABAA receptor agonist muscimol (20mM – 100nL) into the RPa, 20 minutes before injecting the BIC (40 pmol / 100nL) into lateral / dorsolateral PAG. Responses to these injections were accompanied during 40min and compared between groups. Our results were: i) retrogradely labeled neurons were found in all PAG columns; ii) PAG activation by BIC caused positive chronotropism and inotropism, which are accompanied by afterload increases (as evidenced by arterial pressure increases); iii) Inhibition of RPa neurons with Muscimol reduced heart rate, arterial and ventricular pressures. The subsequent injection of BIC into PAG still increased arterial pressure, heart rate and cardiac inotropy outcomes, but the magnitude of these responses was significantly smaller than those evoked by BIC into PAG without inhibiting RPa. PAG neurons project directly to RPa. PAG activation increases cardiac chronotropy and inotropy, and these responses seem to partially rely on medullary ventromedial RPa neurons.