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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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/7410116802190343
Leishmaniasis 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.
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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/7410116802190343
The 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.
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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/3850460442622655
The 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.
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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/7881527576747420
Dengue 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.
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/3512648574555468
The 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.
Acesso aberto (Open Access)
Análise de alterações moleculares nos genes ND1 e ND3 em câncer de pulmão não pequenas células na população paraense
(Universidade Federal do Pará, 2018-03-09) FERNANDES, Lorena Duarte; BORGES, Bárbara do Nascimento; http://lattes.cnpq.br/0676220027193876
Bronchopulmonary carcinoma is the most frequent in the world, being one of the most aggressive neoplasms, with a mortality / incidence ratio of around 90%, with overall survival in five years low, about 10 to 15%, in most populations of the world. In the Northern Region of Brazil, this pathology is the third most frequent among men and the fourth among women. From the anatomopathological point of view, lung cancer is classified into two main types: small cells and non-small cells, the latter being the most incident, accounting for 75% of cases. Currently, the distinction between subtypes is based on histological, immunohistochemical, and molecular differences. In this context, it is important to emphasize that molecular information influences not only diagnosis, prognosis, but also therapeutic behavior. Several genetic and epigenetic alterations of the nuclear genome are related to the pathogenesis of this tumor. However, changes in oxidative phosphorylation resulting from mitochondrial dysfunction have long been suggested as involved in the process of tumorigenesis. Thus, the present study analyzed two mitochondrial DNA (ND1 and ND3) genes belonging to the I complex of the mitochondrial respiratory chain in 66 lung tissue samples from patients with and without non-small cell lung cancer in the population of the state of Pará. the sequencing analysis identified four alterations in the ND1 gene: C3553T, T3552A, C3595ins and G3666A and only two changes in the ND3 gene: A10398G and C10400T. Among the alterations found in the ND1 gene, no statistical significance was observed in relation to the development of lung cancer. However, a structural alteration in the ND1 gene was found in the presence of C3595ins, not yet described in the literature. Whereas, the presence of the A allele, observed in T3552A in the ND1 gene, was significantly associated with a protective effect on the development of lung cancer. Already changes in the ND3 gene (G10398A and T10400C) were significantly associated with lung cancer, these changes in ND3 being potential for use as markers in patients with non-small cell lung cancer
Acesso aberto (Open Access)
Análise de alterações no número de cópias envolvendo os cromossomos 1p e 22 em meningiomas de baixo grau
(Universidade Federal do Pará, 2013-12-13) SILVA, Geanny Pereira da; OLIVEIRA, Edivaldo Herculano Correa de; http://lattes.cnpq.br/0094007714707651
Meningiomas are the second most common type of primary brain tumor, originating in the meninges covering the brain and spinal cord. They show slow growth, and are found more often in the CNS, being benign in most case, although there are also cases of meningiomas classified as malignant. At the cytogenetic level, meningiomas are the most well studied tumors in humans: studies in CNS tumors have shown that most cases had chromosomal abnormalities, and the most common alterations in theis type of tumor are the loss of one copy of chromosome 22 and deletion of the short arm of chromosome 1. These alterations have been associated with the tumorigenesis process, because they are found mostly in low-grade tumors, particularly deletions involving chromosome 22. Thus, the aim of this study was to analyze the occurrence of copy number alterations (CNAs) involving chromosomes 1p and 22 meningiomas grade I and II, and in addition to verifying the existence of other recurrent rearrangements through the application of high resolution comparative genomic hybridization (array - CGH ). Tumor samples were collected from eight patients. All samples showed gains and losses of various chromosomal segments. Except for one case, all others showed, in different degrees though, more deletions than amplifications. Loss of 1p segments was observed in all samples. Some CNAs were recurrent, being found up to six out of the eight cases. Pair 22 showed CNV in all samples, but the total monosomy was observed in only two of the eight samples. The global analysis of CNAs in all samples showed that, although changes 1p and 22 were the most frequent observed alterations, as expected, other genomic regions had also alterations in various samples, indicating a possible involvement of these modifications in the process of tumorigenesis and tumor progression. For instance, alterations in pairs 9, 12 and 17, have been observed in other studies and were correlated with atypical and anaplastic meningiomas. Our data indicate the existence of a larger number of genomic alterations in low-grade meningiomas, disagreeing partly with the assumption that these tumors are characterized by a small number of changes, usually involving pair 22 and, less frquently, loss of 1p. However, the fact that these tumors present alterations that are classically found in meningiomas, even benign, such as deletions in 1p and 22q, may be an indication that these changes must be linked with the early events of origin in meningiomas, as already suggested several times by other authors . In conclusion, these alterations remain important markers in meningiomas, and the relationships of these and other CNAs with the response to different treatments and recurrences should be the next step after cytogenomic characterization based on array-CGH has been completed.
Acesso aberto (Open Access)
Análise de células-tronco adultas (CTA) em cultura de células de tecido epitelial de pequenos roedores (rodentia-stricognathi- sciurognathi)
(Universidade Federal do Pará, 2012-11-13) RISSINO, Jorge Dores; PIECZARKA, Julio Cesar; http://lattes.cnpq.br/6644368250823351
The Adult Stem Cells (ASC) are non-specialized multipotent cells found in the bone marrow, peripheral blood, cornea, retina, brain, muscles, dental pulp, liver, pancreas, skin epithelium, digestive system, umbilical cord and placenta. These cells can indefinably reproduce and renew themselves and, under some stimulation, to change into specialized cells of different tissues or organs. The present work had the aim of obtaining ASC from epithelial tissues from wild rodents of different species (Oecomys concolor – one female, Proechimys roberti – two males, Hylaeamys megacephalus – two males). The methodology for isolation and in vitro culture of epithelial tissue following the previously described protocols, as well as the analysis after cryopreservation of morphology, genome stability, counting and cells viability, clonogenic potential and differentiation on osteocytes, chondrocytes and adipocytes. The ADC were characterized as a homogeneous population of in vitro growing cells adherent to plastic surfaces, which has a morphology similar to fibroblasts and with fusiform shape, with high growing rate and cell proliferation form many successive passages, where the clonogenic assays evaluated the cell renewing. On checking the genome stability on P3, the entire sample had stable karyotypes with the correct diploid number. The methodology for ASC differentiation into osteocytes, chondrocytes and adipocytes cell lines was satisfactory and the cells demonstrated the staining with Alizarin Red S, Alcian Blue and Oil Red O, respectively. The entire sample had capacity of proliferation and differentiation, being a potential source of skin ASC. These species can be used as models for ASC studies.
Acesso aberto (Open Access)
Análise imunológica e genotóxica em Rattus Novergicus da linhagem wistar tratados com ciclofosfamida
(Universidade Federal do Pará, 2016-08-11) CARVALHO, Heleniana Maria Miranda de; BURBANO, Rommel Mario Rodriguéz; http://lattes.cnpq.br/4362051219348099
The development of this work has given up due to the need to better understand the immune system, taking into account the diversity of experimental immunosuppression models as well as the variety of immunological responses and genotoxic differences these, related species, the drug and doses used. Thus, aim of this study was to analyze the effects on the immune system and genotoxic effects in Rattus norvegicus Wistar, after inoculation of the alkylating agent cyclophosphamide (CY). The administration of 50 mg / kg in rodents CY, possible to observe a significant decrease in the parameters of cellularity and relative weight of lymphoid organs. The humoral immunity of rodents has undergone deletion, since the analysis of the antibody titration was performed on the test plate forming cells and hemolysis testing. four inoculations that immunosuppressant and the intervals between the inoculations was determined by recovery of normal levels of the above parameters were performed. Both times the drug was administered, there was a reduction in the number of lymphocytes and neutrophils subsequently decreased, but only the second contact CY was observed immunosuppression. The analysis of the genotoxicity of cyclophosphamide (CY) was analyzed using the comet assay and was of paramount importance because dectamos genomic damage occurring in DNA exposed to different doses of cyclophosphamide (CY), which were 50 mg / kg in the first two phases and 25 mg / kg during the last two phases of the experiment. Furthermore, it was found that the genotoxic effects are cumulative with each CY dose applied, because even being administered in the third phase, the middle concentration (25 mg / kg) of the two inoculations initial CY the damage index does not correspond to half damage indices of the first and second vaccination. However, the analysis and immunologically genotoxicamente rodents, our work will enable testing new therapeutic immunosuppression regimens.
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Análise morfológica in vitro da ação de antifúngicos em cepas de Fonsecaea pedrosoi
(Universidade Federal do Pará, 2014-02-21) MASSOUD JUNIOR, Heleno Ramos; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Choromoblastomycosis (CBM) is a disease caused by traumatic implantation of many species of melanized fungi. The State of Pará is the major endemic area in Brazil and Fonsecaea pedrosoi is the major etiological agent. The treatment is not standardized and many forms of interventions are related in the literature. In the other hand, the in vitro susceptibility test to antifungal drugs may help in the therapeutic choice and in the identification of resistant strains. The objective of this work is to evaluate the in vitro susceptibility of 20 F. pedrosoi clinical isolates to itraconazole (ITZ), ketoconazole (KCZ), fluconazole (FCZ) and terbinafine (TBF) as well as the possible morphological alterations induced by ITZ or TBF in the Minimal Inhibitory Concentration (MIC) and high concentrations. The tests were performed according to the Clinical and Laboratory Standards Institute (CLSI, M38-A2 document) recommendations. The final concentrations of ITZ, TBF and KCZ in each test were to 16 to 0.03 μg/mL. To FCZ the final concentrations were to 64 to 0.125 μg/mL. The MIC was defined as the lowest drug concentration that inhibit 100% the visual growth when compared to the non-treated group after five days of incubation at 30°C. ITZ proved to be the most effective drug in vitro against F. pedrosoi (CIM 90= 1μg/mL). TBF showed a low drug activity with 70% of the isolates with MIC ≥ 0.5 μg/mL. The conidia morphological analysis revealed an increasing in the diameter, an interruption of the cellular division and the formation of little chains after the treatment with ITZ in the MIC. At the high concentration used in the susceptibility test we noticed an irregular shape, a detachment of pigmented material from the cell wall and a vacuolization. Rupture in cell wall and amorphous conidia were observed at 32 μg/mL and 64 μg/mL. Significant alterations were not observed after treatment with TBF at the same concentrations. Moreover, the 5-fluorocytocise (5-FC) and FCZ do not stop the conidia growth at high concentrations. However, ultrastructure alterations were noticed after treatment with 5-FC 64 μg/mL. Thus, it is suggested a different morphological pattern after ITZ or TBF treatment during the in vitro susceptibility test. In synthesis, ITZ shown better in vitro antifungal activity while 5-FC only provoked structures alterations in the highest concentration tested.
Acesso aberto (Open Access)
Análise morfométrica do sistema auditivo periférico da preguiça (Bradypus variegatus)
(Universidade Federal do Pará, 2010-08-27) SOUSA, Pêssi Socorro Lima de; FRANCO, Edna Cristina Santos; http://lattes.cnpq.br/5939607544965550; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170
The mammalian super order Xenarthra is composed of about 31 extant species of armadillos, anteaters and sloths. The tree sloths belong to two genera, Choloepus and Bradypus, which diverged close to 40 million years ago. The similarities between the two taxa, such as the presence of green algae in the fur and the suspensory locomotor ability, are remarkable examples of convergent evolution. The exact location of the xenarthran lineage within the mammalian phylogenetic tree isn’t completely understood yet, with some recent rearrangements of the placental mammal family tree considering xenarthrans to be either most closely related to Afrotheria (that includes shrews, aardvarks, seacows and elephants), or Boreoeutheria (that includes primates, rodents, carnivorans and ungulates). The aim of present work is to describe for the first time the morphological features of both the middle and the inner ear of Bradypus variegatus and compare them to other placental mammals whose data is available in the literature. We used 13 mature postmortem specimens (males and females) and 15 skulls from the collection of the Museu Paraense Emílio Goeldi. Than measurements, techniques were used optical microscopy, scanning electron microscopy and computed tomography. Within the sloths’ phylogenetic tree, the genus Bradypus is positioned as the sister-taxon to all other sloths. Our results show that the morphology of the middle and inner ears of Bradypus variegatus are similar to other mammals with data published in the literature and they present allometric scalation.
Acesso aberto (Open Access)
Análise quantitativa de neurônios imunomarcados para parvalbumina no hipocampo e núcleo magnocelular do istmo em Actitis macularius no período de invernada
(Universidade Federal do Pará, 2020-02) GUERREIRO, Luma Cristina Ferreira; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286; DINIZ, Daniel Guerreiro; http://lattes.cnpq.br/3269424921125406; https://orcid.org/0000-0001-7369-2165
It is already known that parvalbumin (PV) neurons have their number modified in face of social, multisensory and cognitive stimuli, both in mammals and birds. However, nothing is known about its plasticity in long-distance migratory shorebirds during wintering period. Here we investigated in four distinct temporal windows of the wintering period, the plasticity of PV neurons of two brain areas of the spotted sandpiper (Actitis macularius) which includes in its migratory journey multiple stopovers for feeding and resting. We used PV as a marker of a subpopulation of inhibitory neurons and count them in the hippocampal formation (HF) and magnocellular nucleus of tectal isthmus (IMC). Based on previous evidence that HF is involved in learning and memory and social interaction, and IMC is essential for control of head and neck and eyes movements, we tested the hypothesis that PV neurons would increase in HF and remain unchanged in IMC. For this, we used the optical fractionator to estimate cell number. Brains were processed for PV immunostaining, followed by estimates of the number of PV neurons of the areas of interest. As compared with migratory rest 1, PV neurons estimates showed significant increase in the hippocampal formation of premigration group. We suggest that parvalbuminergic neurons proliferation is part of the adaptive changes of the hippocampal circuits involved with the migratory process back to the reproductive niches in north hemisphere.
Acesso aberto (Open Access)
Análises dos genes TP53, PTEN, IDH1 e IDH2 em tumores não gliais do sistema nervoso humano
(Universidade Federal do Pará, 2016-06-17) LOPES, Cleiton Mendes; ANSELMO, Nilson Praia; http://lattes.cnpq.br/6518287721873199
Despite the considerable incidence, studies of genetic changes in gene TP53, PTEN, IDH2 and IDH1, in not glial tumors are rare and, in some cases, nonexistent. Glial tumors are usually not classified as benign and rarely evolve to malignancy, with different classifications, effects and locations. The tumor suppressor genes and response to DNA damage, TP53 and PTEN are among the most commonly mutated gene in human tumors. The genes IDH1 and IDH2 are involved in cell metabolism and also were frequently found mutated in gliomas, melanomas and leukemias, currently being considered as good markers for gliomas. Genetic analyzes were performed in those genes, in order to verify that are associated with the etiology and/or progression of non-glial tumors of Human Nervous System (HNS). SSCPPCR techniques for the amplification of the region of interest and mutational screening of samples for subsequent sequencing were used. We analyzed 37 samples of non-glial tumors (14 schwannomas, meningiomas 3, 4 Medulloblastomas, 2 neurocytomas and 14 metastases of Central Nervous System (CNS). Only the gene IDH1 polymorphisms presented on the SSCP 12 (32.4%) samples, and then subjected to sequencing. However, sequencing reactions were satisfactory in only 5 samples, of the polymorphic, (1 metastasis, meningioma 1 and 3 schwannomas). Analysis of these samples have identified 5 different mutations, one present in all, one transversion T → A in codon 106 of exon 4 of the IDH1 gene resulting in amino acid substitution of threonine by serine. Were also identified other mutations in noncoding regions (intron 4) of gene IDH1 in two of these samples. The mutations found in our study had not yet been reported in the literature. Our results indicate the participation the gene IDH1 in the pathogenesis of these tumors.
Acesso aberto (Open Access)
Aspectos morfológicos comparativos entre neurônios da camada I do córtex visual de duas espécies de roedores: Cavia porcellus e Rattus norvegicus
(Universidade Federal do Pará, 2014-08-20) MOREIRA, Thayana de Nazaré Araújo; SILVA FILHO, Manoel da; http://lattes.cnpq.br/2032152778116209
The layer I has as main characteristic the low number of neurons and a high density of nerve fibers. The morphology of neurons of layer I is still understudied, so that in studies evaluating the morphology of these neurons has not yet reached a consensus on the forms and functions of these neurons. This study evaluated the morphology of neurons in layer I of the visual cortex of two rodent species: Cavia porcellus, popularly known in Brazil as a guinea pig and Rattus norvegicus, which is the rat and the Wistar strain was used, commonly used in scientific research. The guinea pig is a widely studied animal model used in several areas of science. Although this species is well studied, works on layer I of this animal are relatively rare, especially in relation to morphology and electrophysiology of neurons in this cortical region. Research in rats on neurons of layer I are more frequent, both in relation to morphology and electrophysiology. To discriminate the potential for differences in the morphology of neurons in layer I of the visual cortex of the guinea pig and mouse, this study classified these neurons according to the trajectory of their dendrites and dendritic measures analyzed using the technique of intracellular injection of biocytin. After classification of neurons comparisons were made between the same cell types of each rodent. 35 guinea India Dunkin-Hartley variety of short-haired of both sexes aged 4-5 days of postnatal life were used. As for the rats, 30 rats of Wistar variety of both sexes aged 14 to 21 days of postnatal life were used. The animals were anesthetized and had their brains removed, separated hemispheres and sections were made in the coronal plane in the occipital region where is located the visual area of rodents. Slices were maintained in artificial cerebrospinal fluid and then brought to the microscope to inject biocytin and subsequently were fixed and treated for mounting on slides and counterstained with Nissl for better viewing. Neurons found were classified as horizontal, ascending, descending and radial. The receptive field area, total and average dendritic length, total area of the cell body, dendrites number, distance from the pia mater and distribution analysis Sholl: The following dendritic measures were analyzed. Results of the most notable were the extent of dendritic branches and the size of the cell body of neurons of layer I of the guinea pig compared to rat. This suggests that in this species, a larger number of neural microcircuits can be established, and therefore greater metabolic rate justified by the size of the cell body.
Acesso aberto (Open Access)
Ativação do receptor canabinóide tipo 1 (CB1r) previne o estresse oxidativo cerebral e inibe o comportamento tipo agressivo em Danio rerio (Zebrafish)
(Universidade Federal do Pará, 2022-08-17) PINHEIRO, Emerson Feio; SILVA, Anderson Manoel Herculano Oliveira da; http://lattes.cnpq.br/8407177208423247
Aggression is a set of complex actions that involve several factors of a genetic, neurophysiological, hormonal and behavioral nature. Furthermore, the brain redox state can also influence aggressive behavior in different species. Thus, modulators of this process can influence the expression of aggressive episodes, between them is the Endocannabinoid System that acts as the main neuromodulator of the CNS, in addition to exerting an antioxidant effect in different conditions. However, its participation in the modulation of aggressive-like behavior needs to be better understood. In this context, this study evaluated the role of cannabinoid receptor type 1 (CB1r) in brain redox state and aggressive-like behavior in Danio rerio (Zebrafish). For this, 64 animals were subdivided into groups: (a) Control (n=26), (b) ACEA (n=30) and (c) AM-251 (n=12), all treated with the drugs of interest: (a) Vehicle (NaCl 0.9%); (b) ACEA agonist 1 mg/kg; (c) 1 mg/kg AM-251 antagonist. The animals were isolated in pairs, without physical contact for 24 hours, followed by pre-treatment and after 30 minutes of pharmacokinetics, the fights were filmed for 30 minutes, the individuals were identified as Dominant or Subordinate and the brains were collected for evaluation of the state brain redox of these individuals. Our results demonstrate, for the first time, that the activation of CB1r by the ACEA agonist modulates aggressive-like behavior and, consequently, partially interferes with the establishment of social hierarchy in Zebrafish, through a redox-independent mechanism. We suggest, therefore, that acute treatment targeting CB1r is a useful neuropharmacological tool to elucidate the role of CES in social interaction and aggressive behavior, allowing a translation with numerous pathologies that have aggression as a behavioral disorder.
Acesso aberto (Open Access)
Atividade leishmanicida do extrato da raiz de Physalis angulata e sua ação na célula hospedeira
(Universidade Federal do Pará, 2013-05-23) SILVA, Raquel Raick Pereira da; SILVA, Edilene Oliveira da; http://lattes.cnpq.br/7410116802190343
Leishmaniasis is an infectious disease caused by various species of the protozoan parasites of the Leishmania genus. The chemotherapy is the only effective treatment for the disease, but these drugs are, in general, toxics and requires a longer treatment period. Natural products have been used as traditional medicine and offer new perspectives and represent an important source of new antileishmanial agents. Thus, it is of great importance to assess the effects of the aqueous extract of the root of Physalis angulata, a plant widely used in popular medicine, in promastigotes and amastigotes of Leishmania (Leishmania) amazonensis and its effect on the host cell. Physalins D, E, F and G were found present, for the first time, in the P. angulata roots using liquid chromatography/mass spectrometry analysis. Antiproliferative activity and a dose-dependent inhibition of promastigote growth 74.1% and 99.8 % (IC50 35.5 μg/mL), and intracellular amastigotes 70.6% and 70.8% (IC50 32.2 μg/mL) was observed when parasites were treated with 50 and 100 μg/ mL of extract, respectively. The analysis of the microbicidal activity of host cell infected, with L. amazonensis demonstrated that extract is able to reverse the effect caused by the parasite to inhibit the production of reactive oxygen species. This growth inhibition was associated with several morphological alterations assessed by optical microscopy, transmission electron microscopy and scanning such as alteration on cell division, especially in the phase of cytokinesis, alteration in flagellar membrane, in flagellar pocket and duplication of kinetoplast DNA. Already by flow cytometry was possible to confirm that the treatment induced a phosphatidylserine exposure and decreased cell volume of promastigotes treated. In the host cell were observed cytoskeleton alterations, high number of cytoplasmatic projections, increase of cytoplasm, vacuoles and spreading ability. No cytotoxicity towards macrophages was observed. We have demonstrated that aqueous extract effectively promotes antileishmanial activity and clearly demonstrate the induction of apoptosis and ultrastructural alterations in Leishmania parasites. Thus, aqueous extract may represent a promising natural alternative source for a new antileishmanial agent.
Acesso aberto (Open Access)
Aumento da ativação neuronal e de marcação de BDNF após degradação das redes perineuronais em modelo experimental de privação sensorial
(Universidade Federal do Pará, 2016-09-23) AGUIAR, Gisele Priscila Soares de; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170; BAHIA, Carlomagno Pacheco; http://lattes.cnpq.br/0910507988777644
The central nervous system (CNS) has the ability to processing and store information collected from the environment, and modifies and adapt under environmental stimuli diversity. However, It has low regeneration capacity after injury or neurodegenerative disease. Several works are demonstrating cellular and molecular mechanisms implicated in CNS plasticity, such as chondroitin sulfate glycosaminoglycans (GAGs-SC) important components of the extracellular matrix from nervous tissue, responsible for synaptic stabilization, toconcentrateof growth factors and ions around neurons. Removing CSPG of the nervous tissue, we can (re)opens a potential plasticity window in the CNS. The goal of our work is to evaluate the influence of removal of GAGs-SC on neuronal activity, via cFos immunolabeling, and BDNF proteins levels at the barrel cortex, under an experimental model of sensory deprivation (vibrissectomy) during critical period of plasticity. To do that, we used 18 rats (Rattus novergicus), Wistar lineage, submitted to the removal of all whiskers from their right snout (vibrissectomy) since first day of life (P0) until the end of critical period of plasticity (P30). The 40 days deprived animals received epidural polimer implant of Elvax, previously saturated with chondroitinase ABC (ChABC, to degraded the extracellular matrix) or with bovine albumin serum(BSA, control), on the barrel cortex of contralateral cerebral hemisphere to the sensory deprivation (left). The animals were perfused 10 (P50) or 20 days (P60) after Elvax implant. Our results shown that the animals submitted to the sensory deprivation, during critical period of plasticity of S1, and to GAGsSC degradation presents modification in perineuronal net (PNNs) characteristics when compared to control animals, at P50. Those animals also presents increase in cFos labeled cells (mainly at the granular layer of S1) and in BDNF labeled cells at the deprived PMBSF, both seen in 10 (P50) as 20 days (P60) after Elvax implant saturated with ChABC. In this way, we concluded that GAGs-SC removal induced local plasticity, evoking changes in cortical activity and BDNF expression at the deprived PMBSF, even 30 days after critical period of plasticity ended at S1.
Acesso aberto (Open Access)
Avaliação da influência do tratamento com indometacina no aprendizado e na memória espacial em modelo murino de diabetes tipo 1
(Universidade Federal do Pará, 2017-05-25) SANTOS, Gabriel Cardoso de Queiroz; BASTOS, Gilmara de Nazareth Tavares; http://lattes.cnpq.br/2487879058181806
Diabetes mellitus (DM) is the group of metabolic disorders that has as a common characteristic the disregulation of blood glucose levels, invariably leading to hyperglycemia. This disease has become the most frequent in the adult population, mainly in developing countries, causing several serious consequences such as cardiovascular and renal diseases, factors responsible for a high mortality rate of the individuals affected. In addition that consequences, which are better investigated and described in the literature, other types of complications are observed. Clinical and experimental studies demonstrate that both type 1 and type 2 diabetes mellitus may contribute to the development of cognitive deficits and dementias. However, the mechanisms that lead to such disorders are not yet fully understood. A study using the non-selective non-steroidal anti-inflammatory, indomethacin, has shown that aspects related to impaired neuronal plasticity in diabetes can be reversed, demonstrating that these disorders may be modulated by neuroinflammatory changes. The aim of the present study was to evaluate the influence of chronic treatment with indomethacin on memory and learning in a murine model of type 1 diabetes mellitus (T1DM). Using the open field test, Y-maze test and Morris water maze test we investigated the indomethacin effects on behaviors changes after aloxan inducing T1DM. Indomethacin significantly decrease related behaviors to the anxious state in Open field test. This treatment also reversed space work memory deficits in the Y-maze test, and learning and spatial memory deficits in the Morris Water Maze. Thus, it can be concluded that chronic treatment with indomethacin has beneficial effects on the cognition of mice submitted to type 1 diabetes mellitus.
Acesso aberto (Open Access)
Avaliação da seletividade olfatória causada pela infecção por SARS-COV-2
(Universidade Federal do Pará, 2022-09-16) ALMEIDA, José Ramon Gama; BASTOS, Gilmara de Nazareth Tavares; http://lattes.cnpq.br/2487879058181806; ÁVILA, Paulo Eduardo Santos; http://lattes.cnpq.br/4673218055614655
Introduction: Sudden loss of smell is one of the most prevalent symptoms of COVID-19. The sense of smell ranges from detecting warning odors in the environment to building our most pleasurable experiences. This sense stimulates a complex neural network, including the temporal lobe, the amygdala, the insula and a large part of the limbic lobe: the loss of smell should not, only, be considered a sensory symptom, but also a psycho-sensory syndrome. Objectives: This study was aimed to evaluate the olfactory selectivity and trigeminal sensation in olfactory alterations reported by patients diagnosed with COVID-19. Methods: The Randomized case-control study involving 88 individuals: COVID-19 without olfactory dysfunction previously diagnosed prior to the pandemic period; with COVID-19 with olfactory dysfunction diagnosed before the pandemic period; and without COVID-19 and loss of olfactory sensitivity during the pandemic period, with persistent anosmia or hyposmia after SARS-CoV-2 infection. All individuals participating in the study (Control/intervention groups) diagnosed with or without COVID-19 underwent evaluation data collection form and psychophysical sensory test which included olfactory test, olfactory memory test, olfactory threshold test and trigeminal sensation test (CEP-UFPA: 40962420.2.0000.0018). ANOVA follow the Tukey test. Results: The results demonstrated which all patients with or without COVID-19 diagnosis, loss the ability to not identify the odorant of banana essence when compared to the healthy health group being significant control vs. COVID-19 without p<0.0002 and significant control vs. COVID-19 with p<0.0010 and not significant COVID-19 without vs. COVID-19 with p>0.05. For the short-term olfactory memory test which all patients with or without COVID-19 diagnosis, demonstrated an increase in the misidentification of odorants presented when compared to healthy controls, as well as in the olfactory threshold differences in perception were observed between the groups. analyzed. The absence of at least one chemosensory function (cooling sensation) of the trigeminal during the test period was reported by which all patients with or without COVID-19 diagnosis, when compared to the healthy control group. Conclusion: In this way, SARS-CoV-2 infection may be promoting an olfactory dysfunction that affects the perception of banana odor, as well as the affected long-term characteristic such as olfactory memory, olfactory threshold and trigeminal sensation of olfactory loss that COVID-19. 19 generates in patients may provide clues to therapeutic interventions aimed at preventing, alleviating or curing long-term olfactory dysfunction in COVID-19.
Acesso aberto (Open Access)
Avaliação de biomarcadores sorológicos em um estudo de busca ativa de casos novos de hanseníase em área hiperendêmica
(Universidade Federal do Pará, 2016-10-07) GOBBO, Angélica Rita; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Leprosy is a cronic infection diasease clinically characterized by changes in tactile, thermal and painful sensitivity in skin and peripheral nerves. Due to the absence of laboratory diagnosis of leprosy, new tools that contribute for identification of cases are necessary for enable patient treatment before progression to physical disabilities. In this sense, the present study aimed evaluate serological biomarkers contribution for early diagnosis of leprosy. Was perfomed an active case finding study in Mosqueiro district, Belém – Pará. All individuals were clinically examined by experient leprologists doctors and than 5mL of peripheral blood were colleted for future titration of anti-ND-O-BSA, anti-LID-1 e anti-NDO-LID by ELISA. The action of active finding in Mosqueiro district diagnosed 104 new cases of leprosy between 895 subjects examined (11.6%), indicating a high hidden endemy that agree with the high seroprevalence between schoolchildren. Were observed a significant difference among patients with late or early diagnoses, mainly in multibacillary forms. All biomarkers tested showed promising results in detection of late cases, such as related in literature, however, for early cases those molecules identified correctly only 50% of patients. None of biomarker tested presented sufficient sensitivity to detect all leprosy patients, early or lately diagnosed. Besides, LID-1 molecule had evidenced a lower sensitivity for early cases, their high especificity and accuracy suggest their use as a potential tool for serological screening to identify assintomatic subjects with high risk of illness. Thus, we concludes that besides no biomarker had reveled utility as a serological diagnostic tool, the detection of anti-LID-1 presented a possible aplicability as a screening marker of subjects with increased risk to develop leprosy, contributing indirectly for leprosy diagnosis.