Mestrado em Bioquímica

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Agora exibindo 1 - 20 de 66

Caracterização físico-química da isoforma γ-tripsina bovina
(Universidade Federal do Espírito Santo, 2014-06-26) Lacerda, Caroline Dutra; Santos, Alexandre Martins Costa; Santana, Marcos Aurélio de Santana; Ribeiro, Joselito Nardy; Pires, Rita Gomes Wanderley
A novel bovine trypsin isoform was purified from commercial sample by ion exchange chromatography by Sephadex SP C50®. New isoform contain in addition of loss of N- terminus hexapeptide (as found in parent molecule β-trypsin) an intra-chain split between Lys-155 and Ser-156. The novel enzyme denominate gamma-trypsin showed similar properties with α-trypsin isoform in: polypeptide number chain (two chain), molecular masses (23,312 Da), secondary structure, hydrodynamic radius and others. In spite of enzymatic and structural similarities of both isoforms, -trypsin preferably has a lower rate formation from β-trypsin, a lower surface charge, but the gamma-trypsin has a higher thermal stability than α-trypsin. Due to obtaining facility of purification of bovine trypsin isoforms from commercial font, and properties described above, become this enzyme an interesting alternative for the food industry, detergent and biocatalysis research.
Caracterizações bioquímica e hemostática de pacientes com diabetes Mellitus Tipo 2 em insulinização
(Universidade Federal do Espírito Santo, 2014-07-15) Gonçalves, Nadmy Arrivabene Zavaris; Bem, Daniela Amorim Melgaço Guimarães do; Vassalo, Breno Valentim; Gomes, Isabele Beserra Santos
The study aims to characterize biochemically and hemostatic mellitos patients with diabetes mellitus type 2 (DM2) who developed insulin regimen. Composed group under analysis, 40 patients attended the Basic Health Unit of Consolation, Vitória / ES, aged 25 to 80 years, diagnosed with DM2 and that were already evoluted to insulinization. As controls, 40 patients were selected in the same age group without laboratory and / or clinical diagnosis of DM. Markers of inflammation, hypercoagulability and fibrinolysis were measured: Fibrinogen, D-dimer (D-Di) and plasminogen activator type 1 inhibitor (PAI-1). The polymorphism (-675 4G/5G) in the promoter region of the PAI-1 gene was correlated with their serum levels. Biochemical parameters were measured: plasma glucose (PG), glycated hemoglobin (A1C), total cholesterol (TC), HDL cholesterol (HDL-C), LDL (LDLc) cholesterol, triglycerides (TGC), ultrasensitive C reactive protein (hsCRP), urea and serum creatinine. There was still checking BMI, obesity, smoking, hypothyroidism, hypertension, dyslipidemia and insulin resistance. Statistical analysis showed significant differences (p <0.05) between groups with regard to mean values of HDLC, VLDL-C, TGC, urea, hsCRP and fibrinogen. There was significant difference between groups for VLDLc, TGC, creatinine and fibrinogen. Controls present correlations between: fibrinogen and glucose, hsCRP and fibrinogen, PAI-1 and glucose, PAI-1 and BMI. In insulinization DM2 group correlation was observed correlation between: fibrinogen and D-di, hsCRP and fibrinogen, D-Di and glucose (negative), PAI-1 and triglycerides, PAI-1 and BMI. PAI-1 levels were higher in the control group in subjects with genotype 5G5G, 4G5G and 4G4G followed. Binary Logistic Regression confirmed that the variables hypertension and fibrinogen were significant at p-value (0.009) and (0.049) and adjusted odds ratio (4.184, 1.426 to 12.276) and (3.293, 1.006 to 10.775), respectively, showing that hypertensive patients have a risk 4.18 times more likely to have insulinization type 2 diabetes and that individuals with hyperfibrinogenemia have a 3.29 times greater risk. With this study we hope to contribute to better understanding of the complex changes that accompany the user insulinization type 2 diabetic patients in expectation of seeking appropriate treatment and prevention for the macrovascular complications of diabetes.
Alterações comportamentais, bioquímicas e moleculares em modelo animal de inalação crônica de "crack" : papel dos sistemas dopaminérgico e endocanabinóide no córtex pré-frontal
(Universidade Federal do Espírito Santo, 2014-07-17) Azevedo, Lorena Bianchine Areal de; Martins e Silva, Cristina; Pires, Rita Gomes Wanderley; Bittencourt, Ana Paula Santana de Vasconcellos; Ribeiro, Fabiola Mara
abstract
O papel da galalina na modulação da ansiedade experimental mediada pela matéria cinzenta periaquetutal dorsal (MCPD) de ratos
(Universidade Federal do Espírito Santo, 2014-07-18) Soares, Flávia Roberta Chaves; Harres, Vanessa Beijamini; Bortoli, Valquíria Carmin; Aguiar, Daniele Cristina de
Galanin (GAL) is a 29 amino acids peptide that is present in the CNS of many mammals, including human being. The distribution of GAL and its receptors in emotions control structures involved suggests a possible modulatory role of this neuropeptide on anxiety. The dorsal periaqueductal gray (DPAG) is considered a key structure for behavioral and autonomic expression of defensive behavior. However, the role of GAL in this region has not been studied. The DPAG receives galaninergic projections from other structures, but does not synthesize the peptide on their cell bodies. GAL's actions are mediated by 3 metabotropic receptors, GALR1 and GALR3, which increase K+ efflux, and GALR2, which increases Ca2+ intracellular concentration. Using in situ hybridization technique was described the presence of GALR1 and GalR2 receptors in rat DPAG neurons, but there is GALR1 in greater density. The aim of this study was to investigate the involvement of GAL on the modulation of experimental anxiety by DPAG in rats. Therefore, Wistar rats with a unilateral cannula aimed at the DPAG (AP-lambda: 0 mm; L: 2.0 mm; e P: 4.0 mm, 15o), where the drugs were administered, received the following drugs: GAL (0.1; 0.3; 1.0 e 3.0 nmol/ 0.2μL), M617 – selective agonist GALR1 (0.3; 1.0 e 3.0 nmol/ 0.2μL) e AR-M1896 – selective agonist GALR2 (0.3; 1.0 e 3.0 nmol/ 0.2μL). After 5-7 days of recovery, each animal received an injection of drug and tests were carried-out in the plus-maze, elevated T-maze (ETM) or Vogel Test 20 min later. Each experiment was conducted with separated groups of animals (n=5-12). Tests performed at plus-maze after injection of GAL or selective agonists M617 and AR-M1896 into-DPAG did not change percentage of entries and percentage of time spent in the open arms. The analysis showed that treatment with GAL (3 nmol) significantly impaired Avoidance 2 in the ETM, without change Escape behavior. Acute treatment with GAL did not change locomotion in the Open Field. Finally, GAL (1.0 e 3.0 nmol) did not show difference in the number of punished licks at Conflict Vogel Test in comparison with control group. Thus, the anxiolytic effect of GAL in the DPAG seems to depend on the experimental model of anxiety employee and anxiety level generated by them. Key-words: Galanin. DPAG. Anxiety. T-maze.
Avaliação dos efeitos tóxicos da exposição ao inseticida clorpirifós sobre as respostas cardiovasculares e comportamentais em animais experimentais
(Universidade Federal do Espírito Santo, 2014-07-23) Cunha, Alexandre Frinhani; Sampaio, Karla Nívea; Andrade, Tadeu Uggere de; Moraes, Leonardo Resstel Barbosa
abstract
Estudo do potencial efeito neuroprotetor do extrato etanólico obtido da planta medicinal Combretum leprosum em modelo murino da doença de Parkison
(Universidade Federal do Espírito Santo, 2014-07-31) Moraes, Lívia Silveira de; Silva, Cristina Martins e; Santos, Adair Roberto Soares dos; Pires, Rita Gomes Wanderley
The Parkinson’s disease (PD) is a neurodegenerative disturbe caused by neuronal loss of the dopaminergic neurons in the substantia nigra pars compacta, afecting by 1% in people >70 years of age. Currently, the main treatment is based on the replacement of the dopamine levels (DA) with administration of levodopa, which mitigates especially the motor impairment. Other drugs, such as dopamine agonists, are now used concomitantly with levopoda but they are not effective and do not prevent disease progression, beside triggers several side effects. Thus, there are efforts focused on the discovery and identification of new molecules with neuroprotective activity of natural origin, especially of the biodiversity of flora. The plant Combretum leprosum has vast medicinal potential with a wide spectrum of biological action, such as anti-inflammatory, analgesic, antiepileptic, among others, but its mechanism of action and its neuroprotective effects have not been fully elucidated. In this study, we explored the potential neuroprotective effect of ethanol extract (EE) of C. leprosum in a murine model of PD, using the neurotoxin 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) i.p. at a dose of 30 mg/Kg for 5 consecutive days. Exacerbated hyperlocomotion was observed after injection of amphetamine (2 mg/kg) in the group treated with MPTP, an effect prevented by pretreatment with EE at a dose of 100 mg/kg. The deficit in muscle strength induced by MPTP was also prevented by pretreatment with C. leprosum. In biochemical context, treatment with EE was not able to prevent MPTP-induced dopamine depletion in the striatum. However, interestingly, our data suggest that treatment with EE can prevent depletion of dopamine metabolites, homovanillic acid (HVA) and 3,4- dihydroxyphenylacetic acid (DOPAC), in the same region. In addition, treatment with EE prevented the alterations in the metabolism of MPTP-induced DA. Regarding the molecular part, treatment with EE was able to increase the expression of tyrosine hydroxylase (TH) gene in the midbrain region of the animals. Although there is a clear tendency that the lower expression of the dopamine transporter (DAT) and the dopamine D2 receptor in this region caused by MPTP can be prevented by treatment with EE, this finding was not statistically significant. In the striatum, the expression of genes TH, D1 and D2 did not differ between groups. Our results show that treatment with EE at a dose of 100 mg/kg prevents motor and molecular alterations induced by MPTP, however partially reversing the biochemical changes. Accordingly, our study demonstrates the therapeutic potential Combretum leprosum in the prevention and treatment of PD.
Determinação da atividade e da estabilidade termodinâmica da isoforma α-tripsina bovina em meios aquo-orgânicos
(Universidade Federal do Espírito Santo, 2015-02-24) Pereira, Evaldo Vitor; Gonçalves, Juliana Barbosa Coitinho; Santos, Alexandre Martins Costa; Pires, Rita Gomes Wanderley; Fernandes, Patrícia Machado Bueno; Bemquerer, Marcelo Porto
abstract
Neuroproteção mediada pelo fator estimulador de colônias de granulócitos (G-CSF) em modelo de isquemia cerebral global de camundongos hipercolesterolêmicos
(Universidade Federal do Espírito Santo, 2015-02-26) Peruch, Brunelli da Rós; Nogueira, Breno Valentim; Silva, Cristina Martins e; Pereira, Thiago de Melo Costa
abstract
Efeito de diferentes agonistas de receptores galaninérgicos no núcleo dorsal da rafe de ratos expostos a modelos animais de ansiedade e pânico
(Universidade Federal do Espírito Santo, 2015-02-26) Morais, Juliana da Silva; Harres, Vanessa Beijamini; Bittencourt, Athelson Stefanon; Andreatini, Roberto
Galanin (GAL) is a peptide present in the CNS of various mammals, including human being. Three different receptors have been cloned for the GAL, GALR1 and GALR3, inhibitory, and GALR2, excitatory. The distribution of GAL in structures involved in the control of emotions, and behavior studies suggest that GAL may be involved in the neurobiology of anxiety. The effect seems to depend on both the GAL administration site as involved receptor subtype. The dorsal raphe nucleus (DRN) is distinguished by the presence of serotonergic neurons, important to mediate the antidepressant effect of several drugs. About 40% of the neurons of the DRN co-expressing serotonin and GAL. The activation of GALR1 in this structure decreases the firing rate of serotonergic neurons in the DRN. Previous results from our laboratory showed anxiolytic effect of GAL intra-DRN in rats exposed to the elevated T maze (ETM), but no effect on the escape, related to panic. One of the limitations of this model is that the escape (latency to leave the open arm) usually occurs within a few seconds, so that detect panicogenic effect (low latency escape) can be difficult. Thus, one aim of this study was to investigate the effect of intra-DRN rat GAL in another experimental model of panic, electrical stimulation of the dorsal periaqueductal gray matter (DPAG), more sensitive to detect panicogenic effect. Furthermore, given the existence of different subtypes of galaninergic receptors (GALR1 and GALR2) in the DRN with opposite transduction mechanisms, inhibitory and excitatory, respectively, it is possible that activation of GALR1 receptors is responsible for mediating the anxiolytic/panicogenic effect while activation of GALR2 receptors induces anxiogenic/panicolytic effect. Accordingly, we tested this hypothesis using a selective agonist for GALR1 (M617) and a selective agonist for GALR2 (AR-M1896) in the DRN in rats exposed to ETM. The administration of M617 1.0 and 3.0 nmol in the DRN facilitated inhibitory avoidance, suggesting an anxiogenic-like effect., while administration of AR-M1896 3,0nmoles in the DRN impaired the inhibitory avoidance, suggesting an anxiolytic-like effect, both without changing locomotor activity of animals tested in the open field. Also there was no effect of these drugs on the ETM escape behavior. Later, the pre-treatment with WAY100635 was tested in order to verify that administration of a 5-HT1A antagonist would be able to block the effects seen with GALR2 agonist in animals exposed to ETM. The anxiolytic effect of AR-M1896 was attenuated by the prior administration of WAY100635 at a dose of 0,18nmol. Therefore these results suggest a relationship between the observed effect with the AR-M1896, the release of 5-HT and activation of 5-HT1A receptors. And finally, the GAL 0,3nmol intra-DRN increased significantly the jumpping and trotting thresholds of animals submitted to electrical stimulation of the DPAG, suggesting a panicolytic effect. Together, the results show that the GAL in the DRN participates in the mediation of behavioral responses related to anxiety and, less clearly, to Panic Disorder. The results also shows that effect of GAL on anxiety depends galaninergic subtype receptor activated.
Efeitos da exposição a um ambiente enriquecido sobre parâmetros comportamentais, bioquímicos e moleculares em um modelo murino da doença de Parkinson
(Universidade Federal do Espírito Santo, 2015-08-05) Hilário, Willyan Franco; Silva, Cristina Martins e; Pires, Rita Gomes Wanderley; Nakamura-Palacios, Ester Miyuki; Rodrigues, Lívia Carla de Melo
abstract
Efeito da privação de sono paradoxal e do bloqueio da síntese da corticosterona sobre os limiares da reação de defesa induzida pela estimulação elétrica da matéria cinzenta periaquedutal e colículo superior
(Universidade Federal do Espírito Santo, 2015-08-19) Barbosa, Rafaella Vila Real; Bittencourt, Ana Paula Santana de Vasconcellos; Bittencourt, Athelson Stefanon; Harres, Vanessa Beijamini; Santos, Jeyce Willig Quintino dos
Panic Disorder (PD) is a common mental disorder that affects up to 5% of the population at some point in life and is characterized by the presence of recurrent panic attacks (AP). It is a psychopathology that may be affected by sleep deprivation (PS), relation that is still poorly understood. In this context, experimental models of AP and PS are useful tools in investigating this possible correlation, especially motivated by the growing of deprivation of sleep, which has become increasingly common in modern society. This study evaluated the effects of paradoxical sleep deprivation (PSP) in the thresholds of defensive behaviors induced by intracranial stimulation (EI) of MCPD and CS in rats, which is an experimental model of AP, as well as verified the influence of corticosterone on these thresholds. 160 male Wistar rats were used (300g), organized into 4 groups of 40 animals each, as follows: control group (CTR) submitted to EI, but no PSP; Deprivation group (PRV), submitted to EI and sleep deprived for 96 hours; Deprivation Group + corticosterone synthesis inhibition (PRB), undergoing treatment with metyrapone, EI, and sleep deprived for 96 hours, and the control group + corticosterone synthesis inhibition (CTB), undergoing treatment with metyrapone and EI, but without sleep deprivation. After 10 days of intracranial surgical implant of electrode in MCPD, the animals underwent 5 stimulation sessions, as follows: 1st (TRI) considered as a screening session - immediately before the deprivation, 2nd (P48) after 48 hours of deprivation, 3rd (P96) after 96 hours of deprivation, 4th (R48) 48 hours after the end of deprivation, and 5th (R96) after 96 hours of withdrawal of deprivation. The thresholds of the individual curves obtained for defense responses in the various stimulation sessions of CS and MCPD (TRI, P48, P96, R48 and R96) of the rats were compared, as well as a given threshold response curves in different groups (CTR, PRV, CTB and PRB). Furthermore, the levels of corticosterone (CORT) were measured in different sessions of EI and compared in the same group, as well as between the different groups. In the CTR group, all behaviors were equal in all sessions when compared to the TRI, however, in deprived animals (PRV), the threshold for galloping (GLP) was significantly reduced in R48 and R96, without changes in other behaviors. In contrast, in the xxi PRB group, Trotting (TRT) increased from P48, while the GLP has not changed in any EI session. Comparing the groups, Jumping (SLT), Micturation (MIC), Exophthalmos (EXO), Immobility (IMO), Defecation (DEF), Trotting (TRT) and Galloping (GLP), were not altered in function of CORT levels produced due PSP, suggesting that corticosterone does not influence the characteristic defensive behaviors induced by electrical stimulation of MCPD and CS. In addition, these results suggest that the delayed effect of the PSP on the GLP thresholds is possibly due to time-dependent neurochemical mechanisms.
Enriquecimento ambiental como estratégia não farmacológica para prevenção dos efeitos de longo prazo da separação maternal
(Universidade Federal do Espírito Santo, 2015-08-21) Ribeiro, Laisa Barroso; Bittencourt, Athelson Stefanon; Bittencourt, Ana Paula Santana de Vasconcellos; Rodrigues, Lívia Carla de Melo; Pires, Rita Gomes Wanderley
The perinatal maternal relationship has a fundamental importance in the development of healthy neural circuits that remain as mental bequest throughout life. In this sense, adverse events in this period have the potential to induce psychopathology in adulthood, increasing vulnerability to psychiatric disorders and substance abuse. In this study, the Maternal Separation (MS) was performed in male Wistar rats, with the intention to mimic a sustained stress in human childhood. After that, the animals were submitted to the Environmental Enrichment protocol, a nonpharmacological strategy employed during a strong brain plasticity period, in order to prevent the harmful effects of MS. In adulthood, we proceeded to the behavioral tests for evaluate depression, anxiety and alcohol abuse, as well as biochemical tests as the analysis of plasmatic corticosterone, indicative of the reactivity of the hypothalamus-pituitary-adrenal (HPA) axis to acute stress, and analysis dopamine and metabolites in structures involved in brain reward process – the mesocorticolimbic way (nucleus accumbens and prefrontal cortex). Statistical analysis was performed using the Student t test and one-way, two-way, three-way or repeated measures analysis of variance (ANOVA)
Efeitos do isolamento social neonatal, da fluoxetina e do lactato de sódio sobre o pânico experimental induzido por cianeto de potássio ou estimulação da matéria cinzenta periaquedutal em ratos
(Universidade Federal do Espírito Santo, 2015-08-31) Moraes, Erich Antonio; Schenberg, Luiz Carlos; Harres, Vanessa Beijamini; Sampaio, Karla Nivea
Panic attacks (PAs) can be precipitated either by the inhalation of 5% carbon dioxide (CO2) or by the infusion of 0.5 mol/L sodium lactate (LAC) in predisposed patients, but not in healthy subjects or patients with other psychiatric disorders. These and other observations suggested that PAs are "suffocation false alarms". The panic disorder is likewise characterized by the high comorbidity with childhood separation anxiety (CSA). Consequently, the CSA has been considered both as a predisposing factor of panic and as an important factor of the resistance to panicolitics. Preclinical studies of our laboratory showed, on the other hand, that panicolitics attenuate experimental panic attacks to both the electrical stimulation of the dorsal periaqueductal gray matter (DPAG) and the intravenous injection of potassium cyanide (KCN) in doses and regimenssimilar to those used in the clinics. These studies also showed that neonatal social isolation, a model of CSA, supress the panicolitic effect of fluoxetine (FLX, 1-2 mg.kg-1.dia-1, 21 days) on panic-like responses to electrical stimulation of DPAG. Therefore, the present study evaluated the effects of a higher dose of FLX (Experiment-1) as well as of the infusion of LAC (Experiment-2) on PAs to electrical stimulations of DPAG or to intravenous injections of KCN, respectively, in rats subjected to social isolation as neonates. In Experiment-1, adult male Wistar rats subjected either to 3-h daily neonatal social isolation (NSI) or brief-handling fictive social isolation (FSI) throughout the lactation period, were implanted with electrodes into the DPAG and treated with saline (0.9%, SAL) or fluoxetine (4 mg.kg-1.dia-1, FLX4) over 21 days. Although the panic thresholds remained virtually unchanged in SAL-treated FSI rats, they were progressively increased in SAL-treated NSI rats. Moreover, FLX4-treated rats showed higher thresholds that those treated with SAL. However, comparison to baseline thresholds showed that FLX4 had differential effects on FSI and NSI groups, increasing or reducing thresholds, respectively. Although there were no significant differences between ISF and FSI at the end of the treatment with FLX4, the threshold percent changes relative to baseline suggest that FLX4 had effects even facilitatory on panic responses. The resistance of neonatally-isolated rats to FLX4 extended previous studies with smaller doses of FLX. In turn, the results of Experiment-2 showed that intravenous infusion of a clinically effective concentration of LAC (0.5 mol/L) does not have any effect on the escape responses to KCN in rats either virgin or submitted to the FSI or NSI. Although the latter results suggest that panics to both KCN (experimental) and LAC (clinical) are mediated by different systems, conclusions require further experiments with higher concentrations of LAC.
Estudo molecular da doença de Huntington e correlações com as manifestações clínicas
(Universidade Federal do Espírito Santo, 2015-08-31) Pereira, Lorraine Poltronieri; Pires, Rita Gomes Wanderley; Guimarães do Bem, Daniela Amorim Melgaço; Silva, Cristina Martins e; Errera, Flávia Imbroisi Valle
Huntington's disease (HD) is a progressive neurodegenerative disease that leads to motor, cognitive and mental impairment. It is caused by CAG (guanine-cytosine-adenine) trinucleotide repeat expansion in the huntingtin gene, resulting in a mutant form which causes brain damage. A previous study of our research group in an animal model for HD observed changes in gene expression related to dynein and dynactin responsible for cellular traffic and neuronal development. The aim of this study was to relate the molecular diagnosis with clinical manifestations of HD and analyze the expression of axonemal dynein heavy chain gene 6(DNAH6), dynein light chains Tctex-type 1 (DYNLT1) and dynactin 3 (DCTN3) in patients. Participants of this study were divided into control group (n = 12) and group with clinical diagnosis of HD (n = 25). The clinical diagnosis was made by the medical team Clinical Genetics Clinic of the University Hospital Cassiano Antonio de Moraes, UFES (HUCAM / UFES) through the Unified scale for assessment of Huntington's disease (UHDRS). The molecular diagnosis of HD was confirmed in 68% of selected patients. In this study, we observed a negative correlation between the CAG expansion and the age of onset of symptoms. The relationship of disease severity with the overall functional capacity (TCF), as well as motor impairment was statistically significant (p <0.05). The CAG expansion and impairment of motor function are reflected negatively on the independence of patients. There was decreased expression of the gene DNAH6 in HD patients compared to the control group, which was consistent with that seen in the expression of this gene in the striatum of animal model with HD. There was no change to the DYNTL1 and DCTN3 genes. In the study, we consider important the ratio of the molecular diagnosis with the clinical study of UHDRS scale essential to assess the rate of progression of the HD in patients. It also suggests that evaluating the expression of the gene DNAH6, is a possible blood marker for the early cellular changes that precede the clinical manifestations of HD.
Síntese e caracterização de nanopartículas de ouro com resina de Virola oleifera e avaliação do seu efeito antibacteriano
(Universidade Federal do Espírito Santo, 2015-09-02) Milaneze, Bárbara Altoé; Guimarães, Marco Cesar Cunegundes; Olivares, Fabio Lopes; Nogueira, Breno Valentim
Introduction: The use of nanotechnology is changing the way different types of materials are used. Colloidal synthesis is an extremely versatile method, relatively low cost compared to other methods of production of metal nanoparticles (NP's). Their behavior is related to the environment in which it finds itself, stability, chemical composition, degree of aggregation, morphology and size, so modulations at the time of synthesis are crucial for biological activity. Much of the route traditionally used for the production of metal nanoparticles, start from toxic solvents that end up giving rise to harmful wastes to health and the environment. NP's studies have shown that they have great potential as antimicrobial agents, drug delivery, among others. This study aimed to develop a sustainable methodology for the synthesis of gold nanoparticles using as reducing agent Virola resin solution oleifera, and further evaluation of antimicrobial and antioxidant activity. Methods: The gold nanoparticles (AUNP's) were synthesized by oxidation-reduction, based on a factorial design 3² where variables were time and concentration of reducing agent. The characterization was performed with UV-Vis analysis, microscopy, Zeta potential, ICP-MS and IR. The antioxidant potential was evaluated for both the resin and for the AUNP's. The antibacterial activity was tested against Staphylococcus aureus and Escherichia coli DH5α 1117 by microdilution technique in broth, and rapid interaction by shaking, followed by transmission electron microscopy analyzes. Results: The resin ferrule oleifera was able to reduce as efficiently as gold sodium citrate (reducing agent used traditionally). Among the varying only the concentration of the reducing agent it was important to determine the characteristics of nanoparticles. The peak absorbance between 520 and 540nm occurred. Together with such the microscopy imaging data confirm the presence of gold nanoparticles. From Green synthesis had the AUNP's various forms and zeta potential showed that AUNP's were stable. The ICP-MS analysis showed that the concentration and reduced with green gold synthesis was 18 higher than with citrate. The gold nanoparticles had antioxidant potential and were able to inhibit the growth of S. aureus by interaction with the membrane.
Efeito de diferentes tempos de exposição a um ambiente enriquecido na memória de camundongos : uma abordagem comportamental e molecular
(Universidade Federal do Espírito Santo, 2015-12-11) Rohor, Bruna Zanetti; Martins e Silva, Cristina; Pires, Rita Gomes Wanderley; Bittencourt, Ana Paula Santana de Vasconcellos; Rodrigues, Lívia Carla de Melo
The environment is formed by a set of situations around the individual and has influence on all organisms. A stressor environment has negative influences in a subject, but an enriched environment (EE) has positive influences. In animals, the AE is defined as a combination of inanimate and social complex stimuli that can improve motor, cognitive and sensory functions. The EE is composed of different objects of different shapes, sizes, colors, textures, among others. Environmental novelty, voluntary physical activity and social interaction are also part of EE. The EE is capable of promoting increased visual, sensory, cognitive and motor functions. Cognitive improvement, increased synaptic plasticity, adult hippocampal neurogenesis, modulation of gene expression and changes in levels of neurotransmitters and neurotrophic factors are EE-induced alterations already described. However, several EE paradigms are found in the literature, and this variety of protocols produces a range of controversial results on the effects induced by EE.
Ressonância de plasmons de superfície localizada (LSPR) em dispersões coloidais de ouro para detecção de Ocratoxina A
(Universidade Federal do Espírito Santo, 2016-03-14) Pereira, Rayssa Helena Arruda; Guimarães, Marco Cesar Cunegundes; Silva, André Romero da; Guimarães do Bem, Daniela Amorim Melgaço
Ochratoxins metabolites are secreted by the fungal species known as Aspergillus and Peniccillium. They are derived from a substituted phenylalanyl isocoumarin, and ochratoxin A (OTA) the most toxic among the two types of ochratoxin, A and B. Their similarity with the amino acid phenylalanine is the origin of their toxicity, which causes an effect inhibitory in various enzymes whose substrate is phenylalanine. The regulation of tolerable levels of ochratoxin in foods for human consumption and feed for animal consumption was defined in some countries. The European Union, for example, limited the maximum levels of ochratoxin in some foods, such as wine (2 ppb), coffee (5ppb) and cereals (5ppb). In order to meet these limits of detection, simpler analytical methods, safe and fast are being developed to make it accessible to use by unskilled people. In this context, nanotechnology has much to contribute, especially the branch of nanotechnology that uses the optical properties of materials. Those consisting of noble metal nanomaterials have a property capable of translating events of interaction between molecules in a measurable signal, the plasmon resonance surface located (LSPR). Furthermore, the possibility of adding selectivity sensitivity led to the conjugation of biological macromolecules and nanoparticles capable of recognizing specific antigens. In the present study, we used gold nanoparticles (AUNP) synthesized with trisodium citrate to evaluate the OTA detection potential. The nanoparticles were characterized by spectrophotometry in the UV-visible, infrared spectroscopy (FT-MIR), Raman spectroscopy, zeta potential and transmission electron microscopy. Anti-Ochratoxin A antibodies were adsorbed to colloidal nanoparticles. Detection assays occurred in an aqueous medium and the detection limit reached 1.10-7 OTA µg.ml-1.
Efeitos comportamentais da cetamina em ratos expostos ao labirinto em T elevado : possível envolvimento da via de sinalização de BDNF/TrkB na matéria cinzenta periaquedutal (MCP)
(Universidade Federal do Espírito Santo, 2016-04-06) Silote, Gabriela Pandini; Joca, Sâmia Regiane Lourenço; Harres, Vanessa Beijamini; Hott, Sara Cristina; Schenberg, Luiz Carlos
abstract
Modulação do sistema colinérgico pelo sistema endocanabinoide em tarefas apreendidas e reapreendidas : efeito do agonista canabinoide WIN-2
(Universidade Federal do Espírito Santo, 2016-05-11) Ferreira, Tamara Andrea Alarcón; Silva, Cristina Martins e; Pires, Rita Gomes Wanderley; Ribeiro, Angela Maria; Harres, Vanessa Beijamini
The Cannabis sativa plant, commonly known as marijuana, contains compounds named cannabinoids, and the 9-tetrahydrocannabinol (9 - THC) is the most active component. After cloning and characterization of cannabinoid receptors, it was found that they bind to 9 - THC and to endogenous ligands, named endocannabinoids (ECs), capable of modulating multiple neurotransmitter systems, and emerging as important regulators of various physiological brain functions. The effect of endogenous and exogenous cannabinoids in the processes of acquisition, consolidation and recovery of information, both in learning and memory is still controversial. Moreover, there is evidence that the processes of acquisition and consolidation have distinct biological basis with probable involvement of hippocampal and cortical cholinergic system in these differences. In order to understand the molecular basis of cognitive processes that involve the participation of the central cholinergic system by the activation of cannabinoid receptors, we proposed the use of a cannabinoid agonist, WIN 55.212-2 (WIN-2). For this, Swiss mice were treated with WIN-2 at a dose of 2 mg/kg and submitted to testing in the aquatic Morris maze to evaluate aspects of acquisition and consolidation of the task. We observed that the cognitive impairment caused by chronic treatment with WIN-2 is mainly related to short term memory in acquisition of spatial task process, while consolidating remained unchanged. This cognitive impairment in the acquisition may be related to a possible increase in the concentration of 2-AG (2-arachidonylglycerol) and decreased of AEA (anandamide) in the prefrontal cortex. Although the behavioral changes observed in this study are subtle, we verified modulation of cholinergic by endocannabinoid system, since treatment with WIN-2 in the memory consolidation period resulted in a decreased basal release of acetylcholine (ACh) in hippocampus. However, this decrease was not associated with cognitive deficits observed. Therefore, this study confirms the importance of both systems studied in the modulation of cognitive processes and hopefully, in the future, help in developing treatments with pharmacological and non-pharmacological approaches, to seek reduce the cognitive deficits caused by drug abuse, as well as neurodegenerative diseases such as Alzheimer's disease.
Papel da neurotransmissão noradrenérgica da substância cinzenta periaquedutal dorsal na modulação de comportamentos defensivos relacionados aos transtornos de ansiedade generalizada e de pânico
(Universidade Federal do Espírito Santo, 2016-07-19) Carvalho, Johnathan Junior Vaz; Bortoli, Valquíria Camin de; Bittencourt, Ana Paula Santana de Vasconcellos; Aguiar, Daniele Cristina de
The dorsal periaqueductal gray matter (DPAG) is a midbrain structure involved in the mediation of defensive behaviors associated with generalized anxiety disorder (GAD) and panic disorder (PD). There is evidence indicating the involvement of noradrenergic neurotransmission DPAG in the modulation of anxiety, however, there is no evidence for their involvement in panic attacks. In this sense, the objective of this study was to investigate the involvement of noradrenergic neurotransmission DPAG in mediating defensive behaviors related to GAD and PD in the elevated Tmaze (ETM), an animal model that combines the inhibitory avoidance response to GAD and the escape response to PD. For this, Wistar rats were given intra-DPAG administration of noradrenaline (10, 30 or 60 nmol / 0,1μL) or saline and tested in ETM. In addition, we investigated the effect of pre-treatment with intra-DPAG nonselective antagonists of alpha and beta-adrenergic receptors, phentolamine and propranolol, respectively, in effect noradrenaline injection in the same structure. Our results show that intra-DPAG administration of noradrenaline at the highest dose impaired the acquisition of inhibitory avoidance, suggesting an anxiolytic-like effect, but no effect on the escape response in ETM.

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