Dissertações em Neurociências e Biologia Celular (Mestrado) - PPGNBC/ICB

URI Permanente para esta coleçãohttps://repositorio.ufpa.br/handle/2011/2375

O Mestrado Acadêmico pertence ao Programa de Pós-Graduação em Neurociências e Biologia Celular (PPGNBC) do Instituto de Ciências Biológicas (ICB) da Universidade Federal do Pará (UFPA).

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Adenosina modula os níveis extracelulares de glutamato induzido por hiperosmolaridade em cultura de astrócitos hipotalâmicos
(Universidade Federal do Pará, 2016-04-29) BRAGA, Danielle Valente; DINIZ, Domingos Luiz Wanderley Picanço; http://lattes.cnpq.br/9601463988942971; SILVA, Anderson Manoel Herculano Oliveira da; http://lattes.cnpq.br/8407177208423247
Recent studies have shown that glutamate release by hypothalamic glial cells is an important physiological response to hyperosmolarity. Furthermore, previous studies point out an accentuated increase of the adenosine levels in renal interstitial fluid after the intake sodium increases. This study aims to evaluate the possible relationship between the adenosine and glutamate releases in primary cultures of astrocytes exposed to hyperosmolarity conditions. Hypothalamic astrocytes cultures of Wistar rats at the first two days after birth were exposed to hypertonic sodium solution (340mOsm/L) in different times (3, 5, 10 e 15 min). After this stimulus, the incubation medium was harvested and the extracellular levels of glutamate and adenosine were determined by High Performance Liquid Chromatography. In order to evaluate the relationship between these compounds in hyperosmotic conditions, we have used treatment of the cultures with adenosine, with R-PIA (an agonist of the A1 receptor), as well as with glutamate (an agonist of the NMDA receptor). Our results showed a significant increase of the extracellular levels of glutamate after the hyperosmotic stimulus with a peak at 5 min. Similarly, we have seen an increase of the adenosine levels in the incubation medium after 10 and 15 min. The treatment with glutamate induced an increase in extracellular levels of adenosine after 15 and 20 minutes in isosmotic medium. The exposure to the NMDA receptor did not induce the release of adenosine in none of the concentrations utilized. The pretreatment with adenosine and R-PIA A1 agonist blocked the release of glutamate induced by hyperosmolarity. Our results also showed that the effect of the stimulus on the release of glutamate and adenosine is sodium-dependent and presents a specific response for hypothalamic astrocytes, which can be modulated by the adenosine A1 receptor activation.
Acesso aberto (Open Access)
Análise in vitro do potencial antitumoral do conjugado LDE/Paclitaxel comparado à formulação do comercial Taxol sobre linhagem celular C6 de glioblastoma de rato
(Universidade Federal do Pará, 2022-09) ANJOS, Ana Carolina Brito dos Anjos; FRANCO, Edna Cristina Santos; http://lattes.cnpq.br/5939607544965550; https://orcid.org/0000-0003-2909-949X; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
Glioblastoma, also known as grade IV astrocytoma, is one of the most common and aggressive types of tumors in the central nervous system. Among the characteristics of this type of tumor, the following stand out: infiltration of isolated tumor cells in normal brain tissue, cell proliferation, angiogenesis and intense necrosis. Currently, the main therapeutic approach consists of surgical resection followed by radiotherapy and chemotherapy. However, in most cases, the tumor is not well defined, spreading through the brain region, which makes it difficult to fully resection. In addition, the removal of tissue from this region can leave several sequels. Consequently, patients have high rates of recurrence and low rates of survival. Another problem in the treatment of this type of tumor is due to the lining of the blood-brain barrier that restricts the entry of molecules and substances, including drugs. Thus, this project aims to analyze the antineoplastic effects of the association of a nanoparticle called LDE with a structure similar to low-density lipoprotein (LDL) that will act as a carrier of the drug paclitaxel (PTX), commercially known as Taxol®, it is a chemotherapeutic drug whose cell antiproliferative action has been proven in the treatment of other types of cancer, such as breast and refractory ovarian cancers. For this purpose, the mouse glioblastoma cell line C6 was used for performing in vitro analysis regarding the effects of these treatments on aspects of viability, cytotoxicity and cell death by apoptosis, using the ApoTox-GloTM Triplex Assay kit (Promega Corporation), which performs the three previously mentioned analyses, sequentially. To evaluate growth and drug effect on PTX and LDE/PTX treatment groups, approximately 1x106 cells were cultured in 96-well microplates at concentrations of 0.01; 0.1; 1 and 10 μM in the times of 24h, 48h and 72h. The control was not exposed to the compounds, containing only DMEM culture medium. Results obtained after treatments with PTX and LDE/PTX were expressed as mean ± standard deviation and analyzed by one-way (cytotoxicity) and two-way (viability and apoptosis) ANOVA, followed by Tukey's post hoc test. Differences were considered significant when p ˂ 0.05.
Acesso aberto (Open Access)
Avaliação de polimorfismos dos genes Timidilato sintase, Metileno-tetrahidrofolato e Metionina sintase em tumores da mama
(Universidade Federal do Pará, 2019-02-25) DURÁN, Miguel Ángel Cáceres; BORGES, Bárbara do Nascimento; http://lattes.cnpq.br/0676220027193876
Breast cancer (BC) is the most common cancer among women in the world and Brazil, after nonmelanoma skin cancer. Polymorphisms in genes involved in the folate pathway have been associated as possible etiological factors of this disease. Thymidylate synthase (TYMS) codes for the thymidylate synthase, responsible for the conversion of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP). TYMS has a polymorphic tandem repeat in the 5'-UTR region (TSER), which generally contains a triple (3R) or double (2R) repeat of a 28 bp sequence. It is thought that the TSER variants are functionally relevant and are associated with BC risk. Another polymorphism in TYMS is 1494del6 and consists of the variation of a 6 bp sequence (TTAAAG) at position 1494 of the 3'-UTR region. These allelic variants are closely related to the level of expression of the enzyme. Methylenetetrahydrofolate reductase codes for the 5,10- methylenetetrahydrofolate reductase, which regulates the balance between cell methylation and nucleic acid synthesis, providing methyl groups for the conversion of homocysteine to methionine. Within the polymorphisms of MTHFR, the SNPs C677T and A1298C generate a reduced enzymatic activity, affecting the synthesis of nucleic acids and the availability of methyl groups for biochemical processes, which could increase BC risk. Methionine synthase (MTR) codes for the methionine synthase, which catalyzes the remethylation of homocysteine to methionine, an essential amino acid and precursor of S-adenosylmethionine, which is a universal donor of methyl groups involved in methylation reactions, including DNA methylation. The role of this polymorphism in cancer risk is still controversial. The aim of this study was to determine if the polymorphisms of the TYMS, MTHFR and MTR genes increase the BC risk. We worked with 61 samples of patients and 35 controls, it was carried out DNA extraction and purification, PCR amplification of DNA fragments including polymorphisms and their subsequent analysis directly through gel visualization, by PCR-RFLP and/or by automatic sequencing. Genotypic and allelic frequencies were determined and were related to the clinical characteristics of the patients and the molecular type of tumor. An analysis of statistical significance was carried out to evaluate the associations of all the polymorphisms with the risk of developing BC and the clinical characteristics of the patients. It was found that 3R allele of TSER and T and C alleles of C677T and A1298C could be associated to BC, although without statistical significance, and TSER and 1494del6 polymorphisms of TYMS could be related to the risk of developing more aggressive breast tumors, although the association is not statistically significant.
Acesso aberto (Open Access)
Avaliação do perfil da resposta celular observado em indivíduos da espécie Cebus apella expostos ao carcinógeno N-Metil-N-nitrosuréia (mnu) e tratados com o modificador da resposta imune canova®
(Universidade Federal do Pará, 2011-04-19) FEIO, Danielle Cristinne Azevedo; BRITO JÚNIOR, Lacy Cardoso de; http://lattes.cnpq.br/9705670940390281; LIMA, Patrícia Danielle Lima de; http://lattes.cnpq.br/3411620003450812
The Immune response modifier Canova® (CA) is a homeopathic drug indicated for patients with a depressed immune system, once this medicine appears to increase the innate immunity and induce an immune response against multiple and severe pathological conditions, including Neoplasms. This increase of innate immunity is due to its involvement in the proliferation and differentiation of hematopoietic cells and mononuclear differentiation induction in bone marrow cells. The chemical compound N-Methyl-N-Nitrosourea (MNU) is a alkylating agent powerful carcinogenic able to cause mutations, chromosomal aberrations and DNA methylation, that induce the imbalance in the defense system of the cell and thus stop mechanisms related to cellular metabolism. This compound has been widely used in the induction of experimental tumors. The goal of this work is to assess the pattern of hematopoietic cell response in Cebus apella primate species exposed to carcinogenic N-Methyl-N-Nitrosourea (MNU) and subjected to the treatment with the Immune response modifier Canova®, through the analysis of biochemical, haematological, immune and cell cycle. We had used 13 (thirteen) adult animals of the species Cebus apella divided into five main groups: the control (negative and positive) and experimental (composed of three subgroups). The first experimental group received the MNU during thirty-five days, the second received treatment with Canova during three days, and the third received the MNU during 35 days and the end of this period received treatment with Canova during three days. Evaluation of cellular immune response was through immunophenotyping (CD3, CD4, CD8, T, B, NK) and complete blood count, liver and kidney function through biochemical analysis (ALT, AST, GGT, urea and creatinine) and kinetic evaluation of cell cycle by flow cytometry. Analysis of haematological values of hemoglobin, hematocrit and red blood cells of positive control groups performed significantly lower when compared to the negative control groups, demonstrating an anemia, while the increased white lineage cells in these groups is probably due to the animal's inflammatory response to neoplastic process. The increase of leukocytes has also been observed in experimental groups treated with the CA, a fact explained by the action of this drug, which acts as a modifier of immune response. Although it has been reported that CA can act by increasing the number of neutrophils, in this study we did not observe this action of the medicinal product, probably by the short time of treatment. Monocytes were decreased in the group treated with MNU and risen in groups that received CA probably by drug act in the activation of macrophages via stimulation of monocytes. In biochemical analysis, urea and creatinine were changed in the groups that received the MNU acutely, these changes, as well as those found in the analysis of liver enzymes, can be associated with typical symptoms of poisoning by carcinogenic drugs. Analysis of cell cycle kinetics of animals treated with CA, there had been a significant increase in cell G0-G1 as well as the percentage of cells in cellular proliferation stage (stageG2-M). We found that CA had minimized the toxicity of the MNU in certain haematological and biochemical, we had observed their partially skill to modify immune response, as improve white blood cell count, but without changing the pattern of immunological markers. Then CA is able to restore some hematopoietic system components and can act as an adjuvant in chemotherapy treatments.
Acesso aberto (Open Access)
Avaliação psicofísica da visão espacial cromática e acromática de pacientes pós-operatórios de microcirurgia vascular para aneurismas intracranianos do sistema carotídeo
(Universidade Federal do Pará, 2017-07-01) BASTOS, Albedy Moreira; RODRIGUES, Anderson Raiol; http://lattes.cnpq.br/4030747999301402; SILVEIRA, Luiz Carlos de Lima; http://lattes.cnpq.br/9383834641490219
The intracranial saccular aneurysms constitute the main cause of spontaneous meningeal or subarachnoid hemorrhage. The complications derived from the sprouting of such aneurysms hemorrhage, classified as sensorial (acuity and visual field), motor (motility of extra-ocular muscle), and perceptual, are considered as well studied by neurosurgery due to relative similarity between cases. However, little is known about possible postoperative perceptual and cognitive alterations. In the present study, the visual sensitivity of patients in postoperative condition of intracranial aneurysm of the carotid system to achromatic and chromatic spatial stimuli was evaluated. The contrast sensitivity to luminance spatial gratings was measured in eleven spatial frequencies (0,2, 0,5, 0,8, 1, 2, 4, 8, 10, 15, 20, and 30 cycles/degree). The color discrimination ability was measured by a version of the Farnsworth- Munsell test with 85 hues, divided in four series of stimuli presented sequentially, being one serie with 22 stimuli and three series with 24 stimuli. For the measure of color discrimination thresholds, 5 MacAdam’s ellipses were determined and, for each one of them, discrimination thresholds had been measured in 20 directions of the CIE 1976 color space, through the test of Mollon-Reffin. The studied variables were compared with the statistical norms of agematched control subjects. The results show that, individually, there are not significant losses of spatial contrast sensitivity. However, individual losses in color discrimination ability and individual alterations in color discrimination thresholds in all color confusion axis (protanopic, deuteranopic and tritanopic) were detected. Thus, it can be concluded that color vision have sensorial parameters which measurement can be useful in the evaluation of patients postoperative of intracranial aneurysm of the carotid system.
Acesso aberto (Open Access)
Distribuição espacial e temporal das redes perineuronais durante o desenvolvimento pós-natal do córtex pré-frontal
(Universidade Federal do Pará, 2022-04) COIMBRA, Gabriele dos Santos; BAHIA, Carlomagno Pacheco; http://lattes.cnpq.br/0910507988777644; https://orcid.org/0000-0003-3794-4710
The mammalian prefrontal cortex (PFC) is the cerebral cortex area involved in processing several functions as cognition and complex motor control for social interactions. In this PFC area, there is no duration of the time window definition about its critical period of plasticity. One of the potential biological markers for this may be the Perineuronal Nets (PNNs). The present work aimed to examine the developmental time course of PNN formation focusing on the medial prefrontal cortex (mPFC) of rats using histochemistry with Vicia villosa agglutinin. We use 21 male rats Rattus novergicus, wistar lineage, which were randomly divided into seven experimental groups, composed of 3 animals in each group, as follows: group at 7, 14, 20, 26, 58, 75, and 135 postnatal days, respectively. We found that in PFC, PNNs appear at P26 with a small number of Vv+ cells, increasing in total numbers until adulthood. The results of the present study demosntrate the temporal development of PNN formation in the Wistar rats mPFC, and we suggest a time window for the end of the critical period of plasticity in this cortical area (26- 75 postnatal days), there is a progressive decrease in PNNs with immature profile and a concomitant increase in mature PNNs during postnatal development of the mPFC, making this PNNs profile more prevalent at more advanced ages, around 3 months of age, when the animals are already considered young adults.
Acesso aberto (Open Access)
Exposição subcrônica ao metilmercúrio induz danos teciduais, bioquímicos e proteômicos em cerebelos de ratos
(Universidade Federal do Pará, 2018-07-04) MATTA, Pedro Philipe Moreira; LIMA, Rafael Rodrigues; http://lattes.cnpq.br/3512648574555468
Methylmercury (MeHg) represents the most toxic form of mercury, which in chronic intoxications induces motor and cognitive impairment in adult rats. Studies suggest that this metal has a tropistic effect on the cerebellum, however few researches aim to elucidate the mechanisms associated with low-dose MeHg-induced damage in a subchronic exposure model. Thus, the objective of this study was to verify motor, tissue, oxidative and proteomic biochemical alterations induced by subchronic exposure to low doses of MeHg. Fifty six male Wistar rats, 90 days old, were divided into two groups: control group (distilled water) and exposed group (0.04 mg / kg / day of MeHg), both administered via intragastric gavage for 60 days. After the exposure period, the open field and rotarod behavioral tests were performed. Subsequently, the cerebellum from these animals were collected for biochemical analysis, proteomics, mercury tissue deposits, and immunohistochemistry evaluation. Statistical analysis was performed using the Student's t-test, considering a significant value of p <0.05. The proteomic profile was analyzed by the ProteinLynx Global SERVER ™ software (PLGS). Proteins with p <0.05 were considered super-regulated proteins and those with p <0.95 were considered as sub-regulated proteins. The test used was Fisher's exact test with Bonferroni correction. Our results demonstrated a decrease in the motor tests of the animals exposed to MeHg. Open field test showed a decrease in total distance covered and the number of rearing in comparison to the control group with p <0.05. Rotarod test presented a decrease in the time for the first latency to fall and an increase in the number of falls in the MeHg group in comparison to the control with p <0.05. The biochemical evaluation showed an increase in nitrite and lipid peroxidation levels and a reductions of Antioxidant Capacity Against Peroxils radicals (ACAP) with p <0.05. Considering the proteomic profile of these animals, among the 1220 proteins identified, 436 proteins were found exclusively in the control group and 311 proteins exclusively in the MeHg group. Also, 358 proteins were found overexpressed and 115 subexpressed proteins. All proteins interactions were depicted on 3 interactions networks to perform proteomic alterations analysis. In addition, the tissue evaluation showed a decrease in Purkinje cells and NeuN + cells and a smaller amount of IBA1+ cells. Area fraction analysis showed a smaller number of GFAP positive cells, synaptophysins and MBP +. Thus, our results suggest that MeHg intoxication provoked cellular and proteomic damage probably related to induced oxidative stress and also reflecting on motor deficit in behavioral tests.
Acesso aberto (Open Access)
Influências do ambiente e da idade sobre a complexidade morfológica dos astrócitos do giro denteado de camundongos suíços albinos
(Universidade Federal do Pará, 2015-05-14) FÔRO, César Augusto Raiol; SOSTHENES, Marcia Consentino Kronka; http://lattes.cnpq.br/7881527576747420; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
During our previous study (Diniz et al., 2010), mice (Mus musculus) maintained in an impoverished environment that mimicked a sedentary lifestyle from weaning generally performed worse on spatial memory tasks (the Morris water maze) and did not distinguish between old and recent or between displaced and stationary objects in episodic-like memory tests. In contrast, mice maintained in enriched cages for equal time preserved those abilities. These behavioral outcomes were associated with layer-dependent, numerical astrocytic changes. Using the same serial anatomical sections selectively immunolabeled for glial fibrillary acid protein from the previous study, we tested the hypothesis that environmental impoverishment would reduce the morphological complexity of astrocytes, and that such changes would be associated with learning and memory decline. We used three-dimensional microscopic reconstructions and unbiased systematic and random sampling approaches to select astrocytes from the polymorphic, granular, and molecular layers of the dentate gyrus. Cluster and discriminant analysis of three-dimensional astrocytic morphometric features from each layer and experimental group revealed two main morphological phenotypes. Type I astrocytes were more complex than type II; they exhibited larger tree areas, larger tree volumes, more segments, and more vertices. Integrated analysis with previous behavioral findings from the same animals revealed that the reductions in morphological complexity observed in young mice from impoverished and aged mice from enriched environments were observed in both astrocyte types in all layers of the dentate gyrus. We suggest that long-term environmental impoverishment and aging effects on astrocyte plasticity may represent at least part of the circuitry changes underlying learning and memory decline.
Acesso aberto (Open Access)
Novas abordagens evolutivas em peixes da Amazônia: mapeamento de elementos repetitivos como marcadores para estudos em espécies do clado Peckoltia (Siluriformes, Loricariidae)
(Universidade Federal do Pará, 2018-04-30) PETY, Ananda Marques; NORONHA, Renata Coelho Rodrigues; http://lattes.cnpq.br/0860824558237244
Cytogenetic data provide important information on the diversity of Loricariidae, as they corroborate the classification analyzes of the species not described and help in the understanding of inter-intraspecific diversity. However, among the species of the Peckoltia clade, the determination of the number of chromosomes alone does not resolve these questions, since most species exhibit a stable diploid (2n) number. Thus, the use of other chromosomal markers is necessary to clarify the genomic organization of these species and to understand their diversity. The physical mapping of repetitive DNA has been widely used as an important tool in the study of taxonomic and evolutionary problems in fish, as well as to understand the processes of genomic organization and diversification. The objective of the present work was to map ribosomal sites (rDNA) 5S and 18S in Ancistomus feldbergae and five species of Peckoltia: P. cavatica; P. multispinis; P. oligospila; P. sabaji and P.vittata, and discuss the mechanisms of organization and diversification of these sequences. The results of the present study demonstrate that all six species analyzed have a karyotype composed of 52 chromosomes but have divergent karyotype formulas. Nucleolus Organizing Regions (NOR) of the single type were observed in Ancistomus feldbergae, P. cavatica, P. multispinis and P.vittata, while multiple NORs were found in P. oligospila and P. sabaji. Extensive variations in the number and location of 5S and 18S rDNA sites among species were observed. These data indicate that inversions are not the only most important events in karyotype evolution in this group and should be useful in identifying the species studied here. In addition to inversions, transpositions are important evolutionary events involved, at least in rDNA clusters that spread in Peckoltia and probably in other species of Hypostominae.
Acesso aberto (Open Access)
Prevalência e associação da infecção gástrica por Helicobacter pylori e do vírus Epstein-Barr em casos de gastrite na população do Amapá
(Universidade Federal do Pará, 2017-11-03) ALVES, Nélisson Clei Ferreira; BURBANO, Rommel Mario Rodriguéz; http://lattes.cnpq.br/4362051219348099
The epidemiology of HP and of the Epstein-Barr virus (EBV) is worldwide. The prevalence of both carcinogenic agents, in the world human population is about 45%. A recent study suggests that EBV coinfection with HP cagA positive increases the oncogenic potential of this bacterium. The objective of this study was to identify the prevalence of the bacterium HP and of the virus EBV and the association of those pathogens and of the cagA gene in patients with gastritis in the population of Amapá. A descriptive study was accomplished, of the traversal type, where 292 samples of gastric mucous of the patients were analyzed with gastritis submitted to the endoscopy, age group between 14 and 83 years. For detection of HP, Urease test and Polymerase Chain reaction were used; this methodology was also useful to reveal the positive cagA of the bacterium. Additionally, the technique of in situ hybridization was used for detection of EBV and the microscopic analysis that determined the histopathological characteristics of the gastric mucous. Results: The study showed high prevalence of cases of HP in patients with gastritis with a relative frequency of 87,67% of the 292 analyzed cases, a higher incidence, of HP positive cases, in female, 88,27%. The incidence of the cagA gene in samples of positive patients for HP was 72,66%, higher prevalence in female, 75,32%. In the present study 8,59% of the patients were found with viral infection caused by EBV in positive samples for HP with bigger prevalence in male, 9,18%. According to the age group, the study showed higher prevalence of the gene cagA and of EBV in positive patient for HP in the age group between 44 and 54 years, with 23,12% and 36,37%, respectively. Conclusion: Most of the findings of this study are similar to the reports from the literature, however, it is necessary other studies in order to explain if there is or there is no correlation between the infection for HP and EBV in the north of Brazil.

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