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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Item Acesso aberto (Open Access) Ação da ciclosporina A na via de ativação do fator de crescimento de nervo (NGF) em células neurais do SNP(Universidade Federal do Pará, 2016-08-26) JESUS, Jessica Batista de; SENA, Chubert Bernardo Castro de; http://lattes.cnpq.br/8620752020290438; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978A is an immunosuppressive drug with known action on T cells of the immune system used in organ transplantation and autoimmune diseases. In the nervous system, cyclosporin A acts by inhibiting the action of Calcineurin, an important second messenger from pathway of signal transduction Nerve Growth Factor (NGF), resulting in hyperphosphorylation of the nuclear factor of activated T cells (NFAT), and downregulation of NGF, TrkA and other factors that participating in this pathway. The NFAT1-4 family are dependent isoforms of calcineurin, while NFAT5 isoform is independent. It has been demonstrated the neuroprotective role of Cyclosporin A via calcineurin dependent or independent. In this study, we evaluate the action of Cyclosporine A in the PNS system, that could be associated with levels of NGF, TrkA and an independent of calcineurin transcription factor (NFAT5) that interplay the plasticity of neuronal cells derived from Dorsal root ganglia (DRG) maintained in cultures. We use E10 DRG cultures supplemented with medium conditioned E9 Retinal treated with Cyclosporin A for 48 and 72 hours. Cultures enriched neurons were confirmed by calcium imaging method. The action of Cyclosporine A in the neuritogenesis was assessed by bright field microscopy, expression of NGF, TrkA and NFAT5 was performed by RT-PCR, intracellular accumulation of NGF was evaluated by immunofluorescence and the presence of TrkA in neurons. The viability test of the cultures treated or not with the concentrations of 1-40μM Cyclosporine A was performed by MTT method. The results show an increase of NGF levels in mixed cultures, and TrkA receptor and NFAT5 in cultures enriched in neurons following treatment with cyclosporine A. Given the importance of NGF pathway in the development and maintenance of the SNP, the use of Cyclosporin A have activity in the peripheral nervous system cells, which might be used in the clinic with new target for new therapies.Item Acesso aberto (Open Access) Ação da hidroxicloroquina sobre neurônios da retina de embrião de galinha(Universidade Federal do Pará, 2017-03-22) ROSÁRIO, Aldanete Santos; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978Hydroxychloroquine (HCQ) is currently used in the treatment of malaria and autoimmune diseases and others therapeutic purposes. However, this drug is known to cause side effects, including producing visual disturbances, which may be irreversible. The mechanisms that produce these visual disorders are not completely known. HCQ - related retinal toxicity may be due to high metabolic rate, being very susceptible to the action of xenobiotics and oxidative damages. Thus, this work aims to evaluate the effects of the HCQ on retinal cells, as well as their possible mechanisms of cytotoxicity. The model used in this work was of cultures of retina cells from chicken embryo. To evaluate cell viability, mitochondrial activity was measured by MTT. The lysosomal function was evaluated by the incorporation rate of the neutral red dye. The levels of reactive species of general oxygen and superoxide anion were evaluated by the CellROX probe and by Nitro Blue Tetrazolium (NBT) and total glutathione levels were quantified using the Ellman reagent. Viability was tested in mixed cultures (glia and neurons) or enriched cultures of neurons and glia after treatment with HCQ and compared with chloroquine (CQ). Cells were exposed to concentrations of 25μM, 50μM and 75μM for 24 hours. The results show that mixed cultures treated with CQ presented a reduction in viability of 36 and 61% at concentrations of 50μM and 75μM, respectively, whereas HCQ did not alter viability at any of the concentrations tested. However, when cultures enriched with glial cells were exposed to HCQ for 24 hours, the concentration of 75μM had a small reduction in cell viability, while that the reduction in neuronal cells was of 20, 33 and 56% at the concentrations of 25μM, 50μM and 75μM, respectively. Even a shorter treatment time (6 hours) there was loss of viability in retinal neurons. The incorporation of neutral red supravital dye was also altered in neuronal cultures treated with HCQ for 24 hours, with reduction of 19 and 32%, compared to the control for the concentrations of 50μM and 75μM, respectively. HCQ significantly reduced the levels of reactive oxygen species produced by the neuronal cells, mainly superoxide anion, 43, 52 and 61% for the concentrations of 25μM, 50μM and 75μM of HCQ in 24 hours of treatment, respectively. In concentrations of 50μM and 75μM of HCQ for 24 for hours, the levels of total glutathione in neuronal cells presented a reduction of 37 and 53%, respectively. When the glial cell conditioned medium was used in neuronal cells for 6 hours after treatment with HCQ, it completely reversed the drug-induced cytotoxicity. When total glutathione levels were measured in culture of glia treated with HCQ for 24 hours no changes were observed. These results suggest cytotoxic action of CQ in mixed culture of chicken embryo retina cells which is not observed in HCQ treatment. However, HCQ showed cytotoxic action when cells are cultured separately, mainly on neurons, which is reversed by some factor released by glial cells in the extracellular environment, and glutathione is a possible candidate to exert this neuroprotective function.Item Acesso aberto (Open Access) Ação do alcaloide (+)-filantidina sobre o protozoário Leishmania (Leishmania) amazonensis e a célula hospedeira(Universidade Federal do Pará, 2014-08-14) MORAES, Lienne Silveira de; SILVA, Edilene Oliveira da; http://lattes.cnpq.br/7410116802190343Leishmaniasis is an antropozoonotic disease caused by parasites of the genus Leishmania. These parasites proliferate primarily within macrophages of mammals and are responsible for promoting a variety of clinical manifestations, such as cutaneous leishmaniasis (CL) and mucocutaneous leishmaniasis (MCL). The treatment available is chemotherapy, but is limited by toxicity and requires a long term treatment. The study of natural products from plants such as antileishmanial agent currently plays an important role in the search for new drugs for the treatment of leishmaniasis. (+)-phylantidine, is an alkaloid extracted from stem of Margaritaria nobilis of the family Phyllanthaceae. The aim of this study is evaluated the effects of (+)-phylantidine on promastigotes forms of Leishmania (Leishmania) amazonensis and host cell. Antiproliferative activity of promastigotes forms was observed when parasites were treated with 50, 100 e 200 μg/mL of alkaloid for 96 hours, with reduction of 73.75%, 82.50% and 88.75%, respectively when compared with non-treated parasites. In the period of 96 hours it was observed an IC50 of 56.34 μg/mL. Amphotericin B was used as reference drug and reduction of 100% in parasites treated with 0.1 μg/mL was observed after 96 hours. Treatment with the alkaloid promoted important changes in promastigotes that were observed by scanning and transmission electron microscopy. Alterations in cell body, flagellum, kinetoplast, mitochondria, rosette formation, presence of electrodense vesicles suggestive of lipid body and increase in structures like acidocalcisssomes were observed. In the host cell no cytotoxic effect was observed in the macrophages treated with the alkaloid and analysis by scanning electron microscopy showed that the alkaloid promoted an increase in the number of cytoplasmic projections, increased cell volume and spreading. Thus, these results demonstrate that (+)-phylantidine was effective in reducing the growth of the protozoa, without citotoxy effect which may represent a promising natural alternative source for the treatment of leishmaniasis.Item Acesso aberto (Open Access) Ação do metabólito secundário 5-hidroxi-2-hidroximetil gama-pirona isolado de fungos do gênero Aspergillus sobre monócitos humanos in vitro(Universidade Federal do Pará, 2012-06-15) COSTA, Josineide Pantoja da; SILVA, Edilene Oliveira da; http://lattes.cnpq.br/7410116802190343The 5-hydroxy-2- hydroxymethyl-gamma-pyrone (HMP) is a secondary metabolite synthe-sized by some species of fungi from Aspergillus, Penicillium and Acetobacter genera. The HMP has several applications, being used as antioxidant, tyrosinase inhibitor, protective agent against radiation and antitumor. Recently, it was also shown that this metabolite acts as a macrophage activator. However, the effect of HMP in human monocytes is unknown. Thus, the aim of this study was to evaluate the effects of HMP on the cell viability and differentia-tion of human blood monocytes in vitro. Human peripheral leucocytes were obtained from blood bag donated from Fundation Hemocenter of Para State. Cell isolation was performed using HISTOPAQUE® 1077-density-gradient. Monocytes were treated for 24, 48 and 72 hours with 50 and 100 μg/mL of HMP. The ultrastructural analysis of treated monocytes showed spreading ability, high number of cytoplasmatic projections and vacuoles, features that are often observed in activating cells. Immunofluorescence analysis of the expression of surface protein specific for the macrophage (F4/80), demonstrated that human monocytes treated with 50 and 100 μg/mL for 48 and 72 h showed the similar pattern of expression of proteins to that of human monocytes differentiated by macrophage colony-stimulating factor (M-CFS). The viability test used showed that HMP has no citotoxicity effect on human mon-ocytes when treated with 50 and 100 μg/mL of HMP. These results demonstrate a new role for HMP as an immunomodulator agent, inducing the differentiation of monocytes into macrophages.Item Acesso aberto (Open Access) Acuidade visual e matriz extracelular no córtex visual primário: alterações associadas à privação monocular precoce e ao enriquecimento ambiental(Universidade Federal do Pará, 2012-11-08) SILVA, Nonata Lucia Trévia da; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286; DINIZ JUNIOR, José Antônio Picanço; http://lattes.cnpq.br/3850460442622655The aim of the present study is to analyze the influence of enriched environment on the visual acuity and on the distribution of perineuronal nets (PNNs) in the primary visual cortex of albino mice that underwent monocular deprivation during the critical period of postnatal development. Mice at 10th postnatal day, were monocular deprived through right eye-lid sutured (M, n = 16) and the control group animals were not submitted to any cirurgical procedures (B, n = 16). After weaning, on postnatal day 21, animals were subdivided in: standard environment (AP) and enriched environment (AE), constituting the following groups: M.AP, M.AE, B.AP and B.AE. After 3 months, animals were submitted to grating visual acuity tests, perfused and coronal sections of their brains processed for Wisteria floribunda agglutinin to posterior stereological quantification through optical fractionator method. B.AP animals present visual acuity of 0.48 cycles/degree, while those raised in enriched environment (B.AE) present a better performance at visual test, reaching 0.996 cycles/degree. Animals with monocular deprivation had significantly lower visual acuity (M.AP 0.18 cycles/degree; M.AE 0.4 cycles/degree). Stereological quantifications revealed that enriched environment increases type 1 and the total number of perineuronal nets at supragranular and granular layers in both hemispheres of deprived animals (ANOVA, two-ways, p < 0.05) and this difference at granular layer is due to an increase of perineuronal nets mainly at the right hemisphere (ipsilateral to the monocular deprivation). At infragranular layer, M.AE animals presented an increase only at the number of type 1 PNNs in both hemispheres.Item Acesso aberto (Open Access) 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/8407177208423247Recent 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.Item Acesso aberto (Open Access) Alterações da morfologia da micróglia do septo lateral e comportamento semelhante ao ansioso em um modelo murino de inoculação sequencial de VDEN1 e VDEN4: influência do enriquecimento ambiental(Universidade Federal do Pará, 2016-05-05) GOMES, Giovanni Freitas; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286; SÓSTHENES, Márcia Consentino Kronka; http://lattes.cnpq.br/7881527576747420Dengue disease is the major cause of deaths by arbovirus infections in Brazil. In the American Continent, the epidemics seem to be associated to the fact that multiple dengue virus (VDEN) serotypes circulate simultaneously. Despite its epidemiological importance and a century of systematic studies dedicated to understand the disease, its detailed pathogenic mechanisms remain poorly understood. The objective of this study was to evaluate possible influence of environmental enrichment on behavioral changes and microglial morphology alterations in the lateral septum after sequential VDEN1 and VDEN4 intraperitoneal inoculations of infected brain homogenates. To that end, we used adult females ten months old of an immunocompetent albino Swiss mouse strain housed in standard or enriched cages. A single intraperitoneal infection of VDEN1 was followed after 28 days by another inoculation of VDEN4. To enhance clinical signs, a regimen of daily alternated injections of VDEN1 or VDEN4 followed 24 hours later by anti-VDEN2 antibody was applied in the last 7 days. Control animals received equal volumes and regime of inoculation of uninfected brain homogenate. We assessed the behavioral changes using the open field exploratory (OF) and elevated plus-maze (EPM). Infected animals housed in standard cages showed significant decrease in time of exploration of the periphery in the OF and in the time of exploration of enclosed arm in the EPM. Uninfected mice housed in standard cages and animal housed in enriched cages did not show same changes. To check how possible microglial changes could be influenced by acute DENV1 infection, secondary DENV4 infection or the passive anti-DENV4 inoculation, we decided to sacrifice groups of animals after which point of inoculation. To evaluate microglial changes, we did selective immunohistochemistry for microglia and macrophages using anti-IBA-1 antibody (Wako, Japan) and we used tri-dimensional reconstruction to morphometric evaluation. Compared to uninfected, infected mice from standard cages showed significant changes in microglial morphology. We also tested the hypothesis that septal microglia is clustered in subtypes and that DENV infection could change this pattern. We noticed microglia is subdivided in three subgroups in physiological conditions, a more complex pattern, a less complex pattern and an intermediate. After DENV1 or DENV4 infection, we observed changes in this pattern, including the appearance of a high complexity cell, increasing the percentage of complexes microglia. We observed these changes in animals from standard cages, but not in animals from enriched cages. Another interesting data is that environmental enrichment appears to reduce this morphometric changes. Based on the evidences, we suggest that sequential infection with VDEN1/VDEN4 in murine model induced behavioral changes and microglial changes in the lateral septum and EA appears to protect animals against these alterations. Based on these evidences, we suggest that microglial from lateral septum present a heterogeneous pattern of morphology and that DENV infection can induce morphological changes, and alterations in the pattern of subdivision, associated with the increase in the percentage of high complexity cells. In addition, infection can induces behavioral changes detected by EPM and OF tests and environmental enrichment seems to protect against microglial and behavioral changes.Item Acesso aberto (Open Access) Alterações de expressão gênica na linhagem de glioblastoma humano U87 após exposição ao MeHg e HgCl2(Universidade Federal do Pará, 2016-12-02) GOMES, Bruna Puty Silva; OLIVEIRA, Edivaldo Herculano Correa de; http://lattes.cnpq.br/0094007714707651; LIMA, Rafael Rodrigues; http://lattes.cnpq.br/3512648574555468The organic and inorganic forms of mercury have been pointed as important contaminants in several world regions due to its toxicological characteristics. Various studies have reported that the intoxication by methylmercury (MeHg) and mercury chloride (HgCl2) can lead to central nervous system impairment. It is generally agreed that glial cells are important for the mechanisms responsible for cellular protection against the damages caused by the mercury. However, little is known about the influence of the mercury in the cells genome. Hence, in the present study we did a complete mapping of the humam glial cells genetic network after mercury exposition with the aim to indentify the possible genetic alterations that occurred via the organic and inorganic forms of mercury. Our results demonstrated that U87 lineage cells are more sensitive to MeHg exposition when compared with HgCl2 exposition. Using an analysis of the concentration curves the LC50 was obtained from 28.8μM and 10,68μM after 4h and 24h exposition to MeHg and a LC50 of 92.25μM and 62.75μM after the same time periods exposition to HgCl2. Regarding the genic pool, our results have shown that both metal forms led to alterations in the genic dosage where the MeHg exposition was highly influenced by the concentration and time, whereas the HgCl2 exposition seemed have been strongly influenced by the exposition time. In total there were 205 indentified genes with a lower genic dosage and 188 genes with elevated expression, (Fold change > 5) after 4h exposition and 5μM of MeHg, and 204 down-regulated genes; and 180 up-regulated genes after HgCl2 exposition in the same concentration. The analysis after 24h exposition showed 90 down-regulated genes and 3 up-regulated genes after 1μM of MeHg; 116 genes were down-regulated and 66 genes were up-regulated after a 10μM exposition of MeHg. As for the HgCl2, there were 98 down-regulated genes and 73 up-regulated genes for the groups exposed to 5μM of HgCl2; 326 down-regulated genes and 66 up-regulated genes for the groups exposed to 62,75μM of HgCl2. Our dataset suggests that both mercurial forms are able to alter the cell genetic expression profile thus interfering in important signaling paths prone to gives rise to biochemical impairments and glial cells phenotypes.Item Acesso aberto (Open Access) Alterações morfo-funcionais em córtex isquêmico de animais tratados com transplante autólogo de células mononucleares da medula óssea(Universidade Federal do Pará, 2015-10-08) BARBOSA JUNIOR, Mário Santos; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170; BAHIA, Carlomagno Pacheco; http://lattes.cnpq.br/0910507988777644Statistical data show stroke as the second leading cause of death and leading cause of disability among all other diseases in the world. The ischemic stroke (ischemic stroke) accounts for about 87% of incidence of strokes. In ischemic stroke, inflammation acts in restraint of infarction caused by ischemic stroke, and on the other hand the intensity of the inflammatory response in neurodegeneration and consequently influence the functional loss. The autologous cell therapy, mononuclear bone marrow cells, promotes modulation in neuroinflammation, being timely during an ischemic event for reduction of tissue loss and functional. In the present study, we used an experimental model of focal ischemic stroke to assess morphological and functional effects of autologous implant mononucleres bone marrow cells (CMMOs) on the morphological and functional changes related to ischemic stroke. We demonstrate in this study that the autologous BM-MNC in acute or acute and subacute periods of ischemic event, promoted neuroprotection and inflammatory modulation able to rebound in preservation and functional recovery in specific activities. We also show that the treatment enhanced in subacute period, the ischemic event, was able to promote increase in morphological and functional improvements promoted by autologous transplantation in acute period.Item Acesso aberto (Open Access) Alterações oxidativas em portadores da doença de Alzheimer: correlação com os estágios da doença e o quadro cognitivo dos pacientes(Universidade Federal do Pará, 2012-06-22) FUJIHARA, Satomi; FREITAS, Jofre Jacob da Silva; http://lattes.cnpq.br/2023282005842112; LIMA, Patrícia Danielle Lima de; http://lattes.cnpq.br/3411620003450812The survival of the population has increased gradually and it is correlated with quality of health services and general living conditions, being particularly marked in industrialized countries. The increase in life expectancy affects the incidence of age-associated diseases. Aging is considered a risk factor for the development of neurodegenerative diseases like Alzheimer’s disease (AD). One theory on the pathogenesis of AD postulates that neurodegeneration is the result of oxidative stress and damage to vulnerable cerebral tissues. The fact that aging is a risk factor in AD also reinforces the hypothesis of free radicals involvement, because the effects of their actions can accumulate for years. It is an insidious, progressive disease and is characterized clinically by progressive loss of memory and other cognitive functions, as well as behavioral and social changes. The memory deficit is the main and usually the first symptom of AD, especially with deficiency of retention and recall of new information. The present study aims to evaluate if there are changes in oxidative metabolism detected in the blood of patients with AD, and if there is correlation with different stages of the disease and the patients’ cognitive framework. We evaluated 30 patients with Alzheimer’s disease and 28 subjects in the control group attended at Unit Education and Assistance Physical Therapy and Occupational Therapy at the University of Pará (UEAFTO-UEPA). Oxidative metabolism was assessed by measuring the total antioxidant capacity equivalent to Trolox (TEAC) and measuring the levels of substances that react with thiobarbituric acid (TBARS). Neuropsychological tests were also performed on all participants in the study. No significant correlations were observed in the performance of neuropsychological tests to the levels of TBARS and TEAC in patients with AD. It was observed that the total antioxidant capacity (TAC) was significantly decreased in AD patients compared to controls whatever stage of disease, showing a possible relationship between TAC and AD. In the evaluation of TBARS was a trend toward higher concentrations in AD patients than in control group but the difference was not statistically significant, only the moderate stage was significant when compared with the control group. Evaluation of the lipidic peroxidation through TBARS a biomarker would probably not be suitable for Alzheimer's disease.Item Acesso aberto (Open Access) Alterações oxidativas em portadores de doença de Parkinson: correlação com critérios clínicos e estágios da doença(Universidade Federal do Pará, 2012-08-31) DOMINGUES, Mariângela Moreno; KIETZER, Kátia Simone; http://lattes.cnpq.br/7986644672973004; LIMA, Patrícia Danielle Lima de; http://lattes.cnpq.br/3411620003450812Parkinson's disease (PD) is a neurologic disease of the most prevalent. In this disease, have neurodegeneration in the nigrostriatal system with altered neuronal circuitry of the basal ganglia leading to motor impairment characteristic of the disease. The classic symptoms are resting tremor, rigidity, akinesia or bradykinesia and postural instability. The pathogenesis of PD remains unclear. However, it is estimated that the mitochondrial and the development of oxidative stress in the substantia nigra have an important role in this process. The diagnosis of PD is clinical and usually occurs late, when most neurons nigrais is degenerate. Some studies have shown the neuroprotective effect of antiparkinsonian medications, and the earlier introduction of treatment better the long-term prognosis of the disease. Therefore, the peripheral development of markers to assist in early diagnosis is important for to start the treatment time delay the progress of neuronal death. The aim of this study was to verify the existence of alterations in oxidant and antioxidant parameters in blood of PD patients and its relationship with disease stage and clinical criteria. We evaluated 30 patients with PD and 30 individuals without the disease. To evaluate the stage of disease and clinical character were applied Hoehn & Yahr and UPDRS (unified scale for Parkinson's disease) scales in parkinsonian patients. To evaluate the oxidative activity in plasma of individuals, was analyzed by measuring lipid peroxidation products from the action of Reactive Oxygen and Nitrogen (Eron, TBARS) and to evaluate the antioxidant response was made to evaluate the Total Antioxidant Capacity (TEAC). In groups mild DP and moderate DP was found higher TBARS value and lower TEAC value compared to controls and impairment PD (p <0.05), confirming the presence of oxidative stress in the early stages of PD. In this study these parameters proved to be good peripheral markers of oxidative stress, contributing to an early diagnosis of PD.Item Acesso aberto (Open Access) Análise cinemática do mecanismo ação-percepção em seres humanos(Universidade Federal do Pará, 2018-08-31) SANTOS, Narrery Silva dos; GOMES, Bruno Duarte; http://lattes.cnpq.br/4932238030330851; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170We used a manifold reaching paradigm, in two experiments, to evaluate the lateralization of motor control of arm movements. In the first experiment, we compared the kinematics of reaching movements in a manifold reaching paradigm towards a vertical bar performed with the right and left arm. Capture of kinematic data was performed with a Microsoft Kinect camera while the participants remained seated during the task. In a second experiment, participants estimated the endpoint of a third-person’s arm trajectory performing the same reaching task, but with last one-third of the movement trajectory occluded. This study included 14 individuals of both sexes with a mean age of 24.6 ± 3.9 years. The results corroborate the theory of hemispheric specialization of control of upper limb movements, with the left hemisphere being associated with precision movements and the right hemisphere with supporting movements and being more sensitive to the influence of gravity. Importantly, this difference is also reflected in the mental simulation of arm movement.Item Acesso aberto (Open Access) Análise citogenética como bioindicador para pacientes com diagnóstico sugestivo de Alzheimer(Universidade Federal do Pará, 2013-08-02) NEGRÃO, Igor Patrick Ramos; BAHIA, Carlomagno Pacheco; http://lattes.cnpq.br/0910507988777644; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170Alzheimer's disease (AD) is a neurodegenerative disease that causes neuronal death and consequent progressive loss of cognitive functions, reducing the capacity for work, interfering with social relationships and behavior of the patient. Among the diseases that cause dementia, AD is the most frequent nature of the vascular a ratio of 4:1, respectively. In addition to the pharmacological therapies, diagnostic methods assist in the early identification of the disease by helping the pretreatment, thus reduced disease progression. Currently cytogenetic studies have demonstrated chromosomal abnormalities in individuals with AD and may aid in the diagnosis of disease. The aim of this study was to investigate the potential of karyotype analysis of peripheral blood lymphocytes as a diagnostic biomarker of Alzheimer's disease. For this work, we used two groups of women aged 65 or more, one group (10) suffering from AD and other normal group (10). Each subject was submitted to the socioeconomic survey, a cognitive screening test (MMSE) and the Venous blood lymphocyte culture and chromosome analysis. Our results demonstrate that the group of women with AD showed high rate of monosomy and trisomy compared to normal women. Through the study of history via questionnaire, we found the lifestyle of both groups. Compared the relationship of chromosomal abnormalities with the cognitive level of the AD group, we evidenced an inverse trend between the number of monosomy / trisomy and cognitive performance. Another aspect of our analysis was the role of each chromosome linked to AD. Chromosomes 1, 14 and 21 showed no trisomy and verify the frequency of monosomy, each chromosome has frequency below 3 % of aneuploidy, i.e., the chromosomes studied did not have a great importance in chromosomal alterations found in the study.Item Acesso aberto (Open Access) Análise citogenética comparativa em espécies de morcegos da subfamília phyllostominae (chiroptera-phyllostomidae) por citogenética clássica e hibridização in situ Flourescente (fish)(Universidade Federal do Pará, 2011-03-29) SILVA, Natalia Karina Nascimento da; PIECZARKA, Julio Cesar; http://lattes.cnpq.br/6644368250823351Bats are a highly distributed and diversified group.The diversity of feeding habits makes the Order Chiroptera one of the highest successes among mammals, being very important, because of these habits, on the control of insects, on pollination, and on dispersion of seeds of many vegetables. The family Phyllostomidae is the third bigger family on number of species into the Order Chiroptera. Among the neotropical ones, this family is the most numerous, being found in the rainforests of South America, especially in the Amazon region, where there is the highest diversity of bats in the World. In the present work it was analyzed cytogenetically a sample of three species of the subfamily Phyllostominae: Chrotopterus auritus, Trachops cirrhosus and Vampyrum spectrum collected in the Pará and Amazon states. The chromosomal data obtained for Chrotopterus auritus (2n = 28 e NF = 52) and Trachops cirrhosus (2n = 30, FN = 56) are in agreement with the ones described in the literature. For Vampyrum spectrum (2n=30 NF=56) we described for the fist time the banding patterns and FISH (Fluorescent in situ Hybridization). The C-banding technique demonstrated a pericentric pattern of distribution of the centromeric heterochromatin in the three species here studied. The FISH with telomeric DNA probes shown only distal hybridizations in all chromosomes of the three species, while the 18S rDNA proble confirmed the location of the NOR observed by Ag-NOR staining, in the long arm of pair 2 Chrotopterus auritus, in the pair 11 of Trachops cirrhosus and in the long arm of the pair 1 of Vampyrum spectrum. The comparative analysis among the species suggests an extensive chromosomal differentiation, with few chromosome pairs being shared among the three genera. Five whole chromosome pairs were conserved without any rearrangement after the divergence of the three lineages. The comparison among the species shows that C. auritus and V. spectrum have more shared pairs between them than with T. cirrhosus. Our results support the phylogenetic association between C. auritus and V. spectrum and suggest the association of T. cirrhosus with the genus Phyllostomus.Item Acesso aberto (Open Access) Análise citogenética de duas espécies do gênero Hylaeamys (Rodentia: Cricetidae) por citogenética clássica e molecular(Universidade Federal do Pará, 2013-04-05) PINTO, Jamilly Amaral; NAGAMACHI, Cleusa Yoshiko; http://lattes.cnpq.br/8887641213110093Rodents are one of the largest and oldest orders of the class Mammalia. In South America, the order Rodentia compromises about 42% of mammal species, and from this more than 50% belong to the family Cricetidae, which includes the subfamily Sigmodontinae. The genus Hylaeamys is inserted in the tribe Oryzomyini and corresponds to one of 10 new genera proposed for species and species groups within Oryzomys. Hylaeamys is the equivalent of "megacephalus group", and consists of the species H. acritus, H. laticeps, H. megacephalus, H. perenensis, H. oniscus, H. tatei and H. yunganus, distributed in Venezuela, Trinidad, Guyana, Paraguay and Brazil, in areas of the Amazon rain forest, Atlantic rainforest and savannah. This study aims to analyze chromosomal markers in two species of the genus Hylaeamys, providing data to assist in its taxonomic and cytogenetic characterization. Nineteen samples of Hylaeamys megacephalus (HME) and four samples of Hylaeamys oniscus (HON) were analyzed. HME has 2n = 54 and HON, 2n = 52. The results obtained by G- and C-banding and Fluorescent In Situ Hybridization with whole chromosome probes from Hylaeamys megacephalus made it possible to determine the chromosomal characteristics of the species studied, as well as allowing a comparative analysis between them, and in comparison with Cerradomys langguthi, observing homeologies and karyotypic differences. The two species of Hylaeamys differ by a centric fission/fusion rearrangement in which HON shows the association of the pairs 14/19 of HME. This association is shared with CLA with an inversion (19/14/19). This work is an achievement for phylogeny and chromosomal studies on the genus Hylaeamys.Item Acesso aberto (Open Access) Análise citogenética de profissionais de serviços de radiologia clínica expostos à radiação ionizante na cidade de Belém, Pará, Brasil(Universidade Federal do Pará, 2011-05-06) CUNHA JUNIOR, Luiz Raimundo Campos da Silva e; BURBANO, Rommel Mario Rodriguéz; http://lattes.cnpq.br/4362051219348099Item Acesso aberto (Open Access) Análise citogenética em morcegos da família Emballonuridae (Chiroptera) da Amazônia Brasileira através de citogenética clássica e molecular(Universidade Federal do Pará, 2011-04-29) ARAÚJO, Ramon Everton Ferreira de; PIECZARKA, Julio Cesar; http://lattes.cnpq.br/6644368250823351This is the first description of the karyotypes of bats of the family Emballonuridae from the Brazilian Amazon region. The species studied were Cormura brevirostris-CBR (2n=22; NF=40), Rhynchonycteris naso-RNA (2n=22 and NF=36), Saccopteryx canescens-SCA (2n=24 and FN=38) and Saccopteryx leptura-SLE (2n=28 and NF=38), characterized by G-, C-banding, NOR-staining and Fluorescent In Situ Hybridization (FISH). In CBR the karyotypes found had the same diploid number and fundamental number than in literature. FISH with ribosomal DNA probes and Ag-NOR staining showed two NOR places. Hybridization with telomeric probes showed that the sequences were found in the centromeres of all chromosomes but the Y. Using meiotic studies, chromosome banding and FISH with a whole X chromosome probe from Phyllostomus hastatus (Chiroptera, Phyllostomidae) we suggest that the sex chromosome pair of this species is not the one described in the literature. Cells in diploid and diakinesis had a ring conformation with four chromosome pairs, what suggests multiple reciprocal translocations among these chromosomes, a very rare situation in vertebrates and never found in eutherian mammals. The analyses of RNA, SCA and SLE shows that the karyotypes of Emballonuridae are very conservative even when compared with samples collected geographically very far, but the C-banding analyses shows that it can happen intraspecific variations in the constitutive heterochromatin. For the first time the Nucleolar Organizer Regions were described, showing a stained pair of chromosomes on each analyzed species. The FISH with 18S rDNA probes agrees with the Ag-NOR staining. FISH with human telomeric probes showed hybridizations in the distal portion of all chromosomes. These works are Important to understand the biodiversity of bats from the Amazon region, as well as the comprehension of the chromosomal evolution of Chiroptera.Item Acesso aberto (Open Access) Análise comportamental e histológica de um modelo animal da doença de Parkinson em camundongos suíços(Universidade Federal do Pará, 2011-12-29) GARCEZ, Daniela Rosa; YAMADA, Elizabeth Sumi; http://lattes.cnpq.br/7240314827308306Parkinson’s disease (PD) is one of the most common aging-related neurodegenerative diseases, having a clinical presentation featuring classic motor symptoms related to the degeneration of dopaminergic neurons of the substantia nigra pars compacta (SNpc) and dopamine decr ease in the striatum. Animal models of PD are important tools employed by researcher aiming a better understading of pathophysiologic disease mechanisms and for evaluation of potential therapeutic interventions. Such models must mimic some aspect of the disease as for instances, the degeneration of nigral dopaminergic neurons. In this context, the PD model induced by the injection of the neurotoxina 6-hydroxydopamine (6-OHDA) has been widely established in rats but a better characterization in diferent mice strain is lacking, concerning both behavioral changes and the lesion in nigrostriatal system. Such characterization is important so that this model can be reliably used for investigations of therapeutic interventions. The goal of the present study was to improve the characterization of the unilateral 6-OHDA PD model using Swiss mice, through the evaluation of behavioral changes and the effects on the SNpc dopaminergic neurons. In this investigation we have used a single unilateral intraestriatal injection of 6-OHDA, in two different toxin concentrations: 10 µg/2µl e 20 µg/2 µl. Our results have demonstrated that both 6-OHDA concentrations used provoked severe loss of nigral dopaminergic neurons, amounting to 74,5% e 89,5% respectively. This neuronal loss was highly correlated to the apomorphine-induced rotational behavior but not to the ambulation assessed in the open field test. Therefore, intraestriatal injection of 10 µg/2µl or 20 µg/2µl of 6-OHDA, using Swiss mice, reproduce an effective unilateral 6-OHDA PD model that can be reliably employed in experiments aiming to investigate neuroprotective, cellular and/or pharmacological therapies for PD.Item Acesso aberto (Open Access) Análise da ação cicatrizante dos extratos da folha, pecíolo e caule da Montrichardia linifera (Arruda) Schott in vitro(Universidade Federal do Pará, 2020-02-17) BASTOS, Aline Costa; YAMADA, Elizabeth Sumi; http://lattes.cnpq.br/7240314827308306; BASTOS, Gilmara de Nazareth Tavares; http://lattes.cnpq.br/2487879058181806Chronic injuries affect a large portion of society, treatment for these patients has high financial costs and a result that is not satisfactory. Thus, the objective of this work was to investigate whether ethanolic extracts from the stem, petiole and leaf of Montrichardia linifera (Arruda) Schott have healing activity in vitro. For that, the identification of substance classes of the extracts was carried out by HPTLC, the analyzes of: cytotoxicity, in vitro healing, hematoxylin and eosin morphology and immunomarking for BrdU was also performed. This demonstrated the antioxidant activity and the presence of terpenes in the three extracts, and the presence of flavonoids and phenols in the leaf extract. A serial curve was performed with concentrations of 100 to 0.19 μg / ml of the extracts of the stem, petiole and leaf, in times of 24, 48 and 72h, and it did'n present cytotoxicity. From the cytotoxicity test, the concentrations of 0.78, 0.39; 0.19 μg/ml of the three extracts were chosen for the next tests. Thus, the control group in 6, 12 and 24 hours showed a lesion area of 82.08 ± 12.13, 56.14 ± 15.75, 34.34 ± 10.12%, respectively; while the extract of the stem showed lesion area, in the time of 6h, of 66.108 ± 23.85, 66.10 ± 13.13, 64.81 ± 20.42%, respectively; in 12h, 38.86 ± 20.66, 40.45 ± 14.64, 32.29 ± 16.62, respectively; in 24h, 13.48 ± 11.20, 10.67 ± 7.94, 10.15 ± 7.35%, respectively. The petiole extract showed a lesion area, in 6h, 74.02 ± 15.16%, 80.32 ± 22.50%, 75.56 ± 20.09%, respectively; at 12h, 38.86 ± 20.66, 46.79 ± 12.46, 40.98 ± 5.45%, respectively; in 24h, 13.48 ± 11.21, 27.33 ± 13.86, 12.40 ± 7.72%, respectively. The leaf extract, on the other hand, showed a lesion area, in the time of 6h, 73.08 ± 21.35, 72.91 ± 18.19, 67.84 ± 17.89%, respectively; in 12h, 48.76 ± 21.17, 48.02 ± 17.30, 44.54 ± 18.70%, respectively, in 24h, 24.59 ± 14.58, 26.07 ± 16 , 73, 23.75 ± 15.76%, respectively. There was no significant morphological change in hematoxylin and eosin staining. In the quantification of positive BrdU cells, the control group showed an average of 19.778 ± 3.80, while the groups treated with the extracts at concentrations of 0.78, 0.39, 0.19 μg/ml, for the extract of stem, demonstrated mean of 20.222 ± 1.855, 37.889 ± 7.407, 29.778 ± 4.521 positive BrdU cells, respectively, for petiole extract, mean of 20.222 ± 2.587, 20.444 ± 5.077, 24.889 ± 3.551 positive BrdU cells, respectively; for leaf extract, mean of 20,556 ± 3,504, 23,778 ± 5,974, 22,889 ± 3,1798 positive BrdU cells, respectively. Therefore, Montrichardia linifera stem and petiole extract, in small concentrations, demonstrated wound healing activity in vitro.Item Acesso aberto (Open Access) Análise da atividade enzimática de quitotriosidase como um marcador para a malária vivax: abordagens bioquímicas e moleculares(Universidade Federal do Pará, 2010) CRUZ, Cleber Monteiro; SILVA, Luiz Carlos Santana da; http://lattes.cnpq.br/6161491684526382Chitotriosidase was the first described chitinase and its physiologic role is not entirely clear, although many studies have been showed its participation as a component of human immune response. A 24pb duplication on exon 10 of chit1 gene results on RNAm frameshift, leading to a 87 nucleotides deletion. This alteration generates a protein with no catalytic activity at all. This condition is called chitotriosidase deficiency and presents a frequency close to 6% of homozygosis duplication in different ethnical groups. Malaria is an amazon endemic parasitosis caused by protozoaries of genus Plasmodium and causes symptoms as fever, headache and vomit, which leads to a characteristic immune response. The objective of this study was to evaluate the chitotriosidase enzyme behavior in patients suffering of malaria in Pará state and to determine the frequency of 24pb duplication on chitotriosidase gene in a representative sample. Chitotriosidase measurement was made in 100 healthy individual and in 47 malarial patients. The molecular analysis of the 24pb duplication was realized in 100 volunteers trough a protocol which included DNA extraction techniques, PCR and 2,5% agarose gel visualization to verify normal fragments (normal homozygote: 195pb) and the 24pb duplication (mutant homozygote: 219pb; heterozygote: 219pb e 195pb). This study described at first time on scientific literature the chitotriosidase plasmatic levels increasing in patients suffering of malaria vivax compared to healthy individual. No association was observed between parasitemia and plasmatic chitotriosidase levels in malarial patients. Molecular analysis showed a frequency of 72% normal homozygotes, 24% heterozygotes and 4% mutant homozygotes to 24pb duplication. Allelic frequencies were around 84% to wild allele and 16% to mutant allele. No correlation was found between genotype and biochemical phenotype (represented by chitotriosidase levels) on control group.