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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.
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/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.
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/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 hematológicas, bioquímicas e histopatológicas no modelo de malária aviária Gallus gallus por Plasmodium gallinaceum: papel do óxido nítrico
(Universidade Federal do Pará, 2011-07-29) MACCHI, Barbarella de Matos; DAMATTA, Renato Augusto; http://lattes.cnpq.br/6212140983414786; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978
Malaria causes major losses to human populations in the world. Experimental models are needed for a better understanding of the pathological mechanisms of the diseases and the development of new treatments. Chickens infected with Plasmodium gallinaceum constitute an adequate malaria model due to the phylogenetic proximity of this parasite to human Plasmodium as well as similarities in disease manifestation, as cerebral malaria. The aim of the present study was to investigate the role of nitric oxide in avian malaria development in chickens experimentally infected with P. gallinaceum, treated or not with aminoguanidine (AG - nitric oxide synthase inhibitor). Survival, classical hematology, serum biochemistry and pathology was assayed during the development of the disease. The greatest survival was observed in animals treated with AG that also presented higher parasitemia. Decrease in hematological parameters and Mean Corspucular Volume of erythrocytes increase was showed, indicating bone marrow response to anemia. Lymphopenia and thrombocytopenia were detected in infected animals, but not at the same proportion in treated animals. Monocytes, lymphocytes and heterophils showed an increase in size and changes that indicated activation. Thrombocytes were also higher with the infection and with atypical morphology. Treated animals showed fewer lesions in histological sections of brain, liver and spleen, and NO production decreased, principally during high parasitemia, compared to untreated animals. These results characterize the participation of the chemistry mediator nitric oxide in the pathogenesis of malaria in the avian model.
Acesso aberto (Open Access)
Alterações hepáticas por exposição a baixas doses de metilmercúrio em macacos prego, Cebus apella (Linnaeus 1758)
(Universidade Federal do Pará, 2011-09-16) SILVA, Márcia Cristina Freitas da; SILVEIRA, Luiz Carlos de Lima; http://lattes.cnpq.br/9383834641490219
Cebus apella were exposed to 1,5 ppm methylmercury (methylHg) in the diet for 120 days. Hepatotoxicity was investigated, concentrations of mercury in total blood were monitored each 30 days using atomic absorption spectrometry with cold vapor Hg201, aspartate transaminase (AST), alanine transaminase (ALT) and total bilirubin (BT) were determined. Liver was fixed by formaldehyde 10% and prepared by histopathology protocols. Significant difference was observed in groups exposed and control about total mercury (Hgtotal) in the periods of 60, 90 (P < 0,05) and 120 days (P < 0,01). The histopathology revealed moderate steatosis and hydropic degeneration, common in methylHg exposed in other species. No Significant difference between the levels of AST (p= 0.38), ALT (p= 0.83) and BT (p= 0.07) in groups exposed and control. The Pearson correlation with Hgtotal was negative (AST r= -0,7; ALT r=0,07; BT r= -0,3 e p > 0,05), suggests another studies to clarify the alert levels of mercury concentrations and liver dosages.
Acesso aberto (Open Access)
Análise comparativa dos padrões neurodegenerativos da substância cinzenta em diferentes áreas corticais de ratos adultos submetidos à lesão isquêmica focal
(Universidade Federal do Pará, 2012-09-27) SANTOS, Enio Maurício Nery dos; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
Stroke can occur in any region of the central nervous system (CNS). The cerebral cortex is one of the most often affected areaby this acute neural disorder, but there are no studies that have compared the damaging pattern in different cortical regions after acomparable focal ischemia. The aim of this investigation was to evaluate the degenerative pattern of different cortical areas after focal ischemic injury. Focal ischemia was induced by stereotaxic microinjections of endothelin-1 (ET-1) into the somatosensory, motor and association cortices of adult rats (N = 45). The control animals were injected with the same volume of sterile saline (N = 27). The animals were perfused 1, 3 and 7 days after the ischemic event. The brain was removed, postfixed, cryoprotected, and sectioned in a cryostat. The general histopathology was evaluated in 50μm sections stained with cresyl violet. 20μm sections were submitted to immunohistochemistry for astrocytes (anti-GFAP), activated microglia / macrophages (anti-ED1) and overall microglial population (anti-Iba1). The damaging patterns werequalitatively evaluated under optical microscopy and quantitatively by counting the number of cells in the ipsilateral and contralateral sides to injury.Descriptive statistics and comparisons within and between groups were performed using analysis of variance with Tukey post-hoc test. Conspicuous ischemic tissue loss, microglial activation and astrocytosis were observed mainly 3 and 7 days after ischemia, which was not observed in control animals. The tissue loss and activation of glial cells were more intense in the somatosensory cortex, followed by the motor cortex. The association cortex displayed less damage compared to other cortical areas, which was confirmed by quantitative analysis. The results suggest that an ischemic lesion of the same intensity induces a differential pattern of tissue loss and neuroinflammation, depending on the cortical area, and that the primary sensory and motor areas are more susceptible to ischemia than association areas.
Acesso aberto (Open Access)
Análise evolutiva da morfologia e ecologia em espécies continentais de lagartos do gênero Anolis daudin 1804 (Squamata : Polychrotidae)
(Universidade Federal do Pará, 2007-10-02) PINTO, Gabriel Silva; ÁVILA-PIRES, Teresa Cristina Sauer de; http://lattes.cnpq.br/1339618330655263
Acesso aberto (Open Access)
Análise morfológica e topográfica das células ganglionares da retina do caititu (Tayassu tajacu)
(Universidade Federal do Pará, 2014-12-22) COSTA, Kelly Helorany Alves; ROCHA, Fernando Allan de Farias; http://lattes.cnpq.br/3882851981484245
In the Amazon there are several different wild animals species, becoming this way a good environment to investigated about compared physiology. Among this species, the caititu, Tayassu tajacu, stood out. This animals is located in central America and Latin America. There are many publications around morphology of sexual organs, meat and blood of the catitu. Although, regarding studies about visual’s morphology are scarce. Facing this reality, the present study investigated a morphology and topography of ganglion cells of Tayassu tajacu’s retina. Were used six retina, from eight animals, Both sexes of the species Tayassu tajacu. The caititu born and raised in captivity in the Brazilian company of research known as Embrapa/Pará. Were slaughtered according to the rules of animals ‘s management to subsequent removal and attachment of eyes. The retinas were dissected and stained using Nissl technique to ganglion cells of view, displaced amacrine, red blood cells, microglia cells and components of vascularization. A count of ganglion cells was made a long the horizontal and vertical axis. Being the number of ganglion cells by field converted into density values. The different regions of the retina were analyzed as the cellular density, obtaining the medium value of the density 351,822 ± 31,434 CG/mm². Verified different of the density between the studies regions : The dorsal region had a medium density and standard deviation 894 ± 44 CG/mm², the ventral region 894 ± 1 CG/mm²; the nasal region 1.403 ± 43; the temporal region with 1596 ± 251. The average peak density located approximately 3.13 mm from the dorsal direction and 6.77 mm in the temporal direction of the optic nerve, was 6767 GC / mm². Check there are two specialized regions, the visual streak region and the area temporalis. The visual streak located in the horizontal direction of the nasal region to temporal, presenting higher cellular density, possibly providing better panoramic vision from the environment and detecting the objects in the horizontal direction. Already the temporalis area, located within the visual range, provides increased visual acuity and spatial resolution, the environment that they live. These results allow to start comparisons between morphophysiological the retina of peccaries with other animal species.
Acesso aberto (Open Access)
Aspectos reprodutivos de Anableps anableps (Linnaeus, 1758) (Cyprinodontiformes: Anablepidae) no Estuário Amazônico
(Universidade Federal do Pará, 2010) OLIVEIRA, Valéria de Albuquerque; MONTAG, Luciano Fogaça de Assis; http://lattes.cnpq.br/4936237097107099
Acesso aberto (Open Access)
Aspectos reprodutivos e alimentares da piranha Serrasalmus gouldingi fink & machado-allison, 1992 (Characiformes: Serrasalmidae) em rios afogados da Amazônia Oriental
(Universidade Federal do Pará, 2012) PRUDENTE, Bruno da Silveira; MONTAG, Luciano Fogaça de Assis; http://lattes.cnpq.br/4936237097107099
The present work aimed to evaluate the reproductive biology, condition factor and feeding ecology of Serrasalmus gouldingi in relation to fluviometric variations of lower Anapu River, in “drowned” rivers of Caxiuanã National Forest, Eastern Amazonia, Pará, Brazil. 275 specimens were bimonthly collected during July 2010 to May 2011 and their total length, total weight, gonad and stomach weight were measured. The gonads were analysed histologically to verify the sex and maturation degree, while the stomachs were evaluated through the identification of the consumed items. The sexual proportion did not differ from 1:1considering the studied period as a whole, however the females were more frequent during the transitional periods of drawdown and filing. The species spawning was “parcelada”, showing two peeks of reproductive activity during the periods which preceded the increase in the local fluviometry. The L50 was estimated in 12.24cm for males and in 16.13cm for females. The species growth was positively alometric, showing a gain of weight in relation to length, and the condition factor, when analysed along the entire sample period, decreased mostly during the spawning. The diet of S. gouldingi was composed of 32 items, grouped in 10 categories. The composition indicated an omnivorous diet with a strong tendency to piscivory, once fish fragments were the most predominant item, followed by fruits and seeds and allochthonus arthropods. The distinct hidrological periods were significantly different in relation to diet composition, although no significant difference was detected among sex and maturation degree. Males and females showed the same variation pattern of repletion index, with a more intense feeding during the increase of the river´s level, while it was less intense during the transitional periods (drawdown and filing). The species also showed some variation in their breadth niche, with lower values during the wet period, attributed to the almost exclusive consumption of fruits and seeds.
Acesso aberto (Open Access)
Ativação microglial, lesão da substância branca e expressão de Nogo-A em ratos submetidos à isquemia estriatal
(Universidade Federal do Pará, 2012-05-10) LIMA, Rafael Rodrigues; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
The objective of this investigation was to evaluate the degenerative pattern of several white matter tracts after striatal ischemic injury, correlating degenerative process standards with the microglial activation and expression of Nogo-A. For this purpose, focal ischemia was induced with stereotactic injection of endothelin in striatum of adult rats, and only in the control animals injected with sterile saline. The animals were perfused 3, 7, 14 and 30 days after ischemia. The brain removed, postfixed, cryoprotected, cut into cryostat sections obtained and submitted to immunohistochemical investigation with the following antibodies: anti-GFAP (1:2000, Dako), anti-Tau-1 (1:500, Chemicon), Anti-MBP (1:100, Chemicon International), Anti-Nogo-A (1:100, Invitrogen), Anti-Iba1 (1:1000, WAKO), ED1 (1:500, Serotec) and Anti-MHC II (Abcam 1:100), besides the viewing of the damage pattern with cresyl violet. Slides are marked by different methods were evaluated qualitatively and quantitatively also some (Anti-Nogo A, anti-ED-1, anti-MHC-II and anti-tau-1), counts were carried out in the striatum and in the corpus callosum. The data were tabulated, statistically analyzed by Tukey test (p <0.05) and micrographs taken of the findings more representative. The slides were stained with cresyl violet revealed an increase in cell density by the infiltration of inflammatory cells to the ischemic area, with a significant increase on day 7. The blades immunostained for GFAP was found progressive increase of the population of astrocytes and an increase in cell volume 7 and 14 days. Oligodendrocyte pathology marked with Tau-1 had peak marking the 3rd day in the striatum and the 7th day in the corpus callosum, and loss of myelin compaction identified by MBP was better at 14, in the different treatment. The microglial activation identified by different immunoblots showed a peak on day 7, both in striatum and in the corpus callosum, but in the corpus callosum with a much smaller number compared to the striatum. The morphology of microglial underwent changes, which found the branched phenotype in control animals, as well as in early and late times after ischemia and amoeboid default / phagocytic day 7, coinciding with the largest number of activated cells. The count of Nogo-A + cells peaked at 3 days observed in the striatum, and there were no differences in the corpus callosum expression Nogo-A 3 to 14 days, only a decrease compared to 30 days. Thus, microinjections of ET-1 induced conspicuous striatal tissue loss, concomitant with progressive microglial activation, astrocytosis, loss of immunoreactivity for myelin basic protein and oligodendrocytes damage in various survival times after focal ischemia. These events affect a few SB tracts, as the corpus callosum. The establishment of the temporal evolution of these events is the neuropathological basis for future studies, in which they should handle the inflammatory response in order to minimize these tissue changes.
Acesso aberto (Open Access)
Atividade antiinflamatória e neuroprotetora da Edaravona no córtex sensóriomotor primário de ratos adultos submetidos à isquemia focal experimental
(Universidade Federal do Pará, 2014-02-12) ARAÚJO, Sanderson Corrêa; BORGES, Rosivaldo dos Santos; http://lattes.cnpq.br/4783661132100859; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
Stroke is a neural disorder originated from blood flow decreasing or interruption, making inadequate energy supply in the region, thus promoting tissue damage. The stroke can be divided in hemorragic or ischemic. The ischemic stroke is more prevalent and can occur through thrombosis or embolism. The ischemic pathology has multiple interrelated events like excitotoxicity, peri-infarct depolarization, oxidative and nitrosative stress, inflammation and apoptosis. An element of fundamental importance in ischemic pathology is the microglial cell, whose activity is closely linked to the progression of environment harm. A therapeutic alternative in the treatment of stroke is a pyrazolone called Edaravone. This study evaluated the neuroprotective effect of Edaravone dose of 3mg/kg in primary sensorymotor cortex after focal ischemic lesion. Edaravone treated animals (N = 10) and animals treated with saline solution (N = 10) in the survival time of 1 and 7 days after the ischemic event was evaluated. Treatment whith edaravone showed by histopathological analysis with cresyl violet a reduction of 49% and 66% in infarct size in animals in survival time 1 and 7 days respectively. Immunohistochemistry studies for microglia/macrophages assets (ED1+) demonstrated a reduction in the presence of ED1+ cells in 35% and 41% survival times for 1 and 7 days, respectively. Neutrophils (MBS-1+) were reduced to 64% only in animals with survival times a day. Harmful patterns were assessed qualitatively and quantitatively. Data was tested by ANOVA with Tukey post hoc test. Differences were considered significant at p < 0,05.
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)
Cinética da infecção pelo arbovírus piry em modelo murino: a resposta do hospedeiro adulto
(Universidade Federal do Pará, 2011-09-30) SANTOS, Zaire Alves dos; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
In the present report, a member of a group of RNA South American viruses found in Brazil, that causes febrile disease in humans and encephalitis in neonate and adult murine models, was selected as a model to study encephalitis outcomes in adult albino Swiss mice. In mice housed under standard conditions with free access to water and food, we induced viral encephalitis by intranasal inoculation of Piry virus–infected brain homogenate and correlated neuropathological features. We quantified the cellular inflammatory response in the septal region using a stereologically based unbiased method with clinical signs and neuroinvasion, comparing the outcomes with those of animals inoculated with uninfected brain homogenate. Three-month-old female mice maintained in standard environment received an equal volume of Piry virus infected or normal brain homogenates into the nostrils. From the 1st to 8th days post-instillation (dpi), five subjects from the infected colony were fixed and processed to detect viral antigens and microglia. Control subjects were sacrificed in the 5th dpi and processed for the same markers. After Piry virus encephalitis induced microglial activation and neuroinvasion of glial cells and neurons mainly in the olfactory pathways early in the disease (2 – 4 dpi), but also included hippocampus, cerebellum and brain stem nuclei later on (5 - 8 dpi). The correlation of the host cellular inflammatory quantitative response in the septal area with clinical signs and neuroinvasion, revealed that the number and the morphology of microglias changed early in the disease before neuroinvasion had reached the septal region and clinical signs had appeared. Great variability in clinical symptoms intensity and survival rate were found in the outbred albino Swiss mice strain as compared with previous report in the inbred C57Bl6 strain suggesting less isogenic background. Taken together, our previous and present report dedicated to investigate Piry virus encephalitis progression in the outbred albino Swiss mice strain may open a new field of investigation of the genetics, anatomical and immune substrates of tropical sublethal arbovirus encepahlitis.
Acesso aberto (Open Access)
Dimorfismo sexual da espessura da retina: uma análise de aprendizagem de máquina
(Universidade Federal do Pará, 2022-03) FARIAS, Flavia Monteiro; SALOMÃO, Railson Cruz; http://lattes.cnpq.br/9518575270670446; SOUZA, Givago da Silva; http://lattes.cnpq.br/5705421011644718; https://orcid.org/0000-0002-4525-3971
The present research compared the accuracy of machine learning algorithms in classifying the thickness and volume measurements of retinal layers as obtained from male and female subjects. The study evaluated the retina of sixty-four healthy participants (38 women and 26 men), with normal vision and without eye or systemic diseases, aged between 20 and 40 years. The data acquisition was obtained with a Spectralis HRA+OCT tomograph in the macular region of the retina and its layers: retinal nerve fiber layer (RNFL), ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), retinal pigment epithelium (RPE), inner retina (IRL) and outer retina (ORL). The classification accuracy was obtained with the following algorithms: support vector classifier (SVC), logistic regression (LR), linear discriminant analyses (LDA), k-nearest neighbors (kNN), decision tree (DT), gaussian naive bayes (GNB) and random forest (RF). The characteristics attributed to each participant's samples were the thickness values in the nine regions of the macula plus the total macular volume of each retinal layer. The statistical tests Two-way ANOVA and Tukey HSD post-hoc were used in the statistical comparisons between the accuracies for the classifier and retinal layer variables, considering a significance level of < 0.05. All factors (classifier, retinal layer, and their interactions) had significant influences on accuracy (p < 0.05). The main effect of the algorithm type factor resulted in an F ratio of F (6, 630) = 4.527, p = 0.0002. The main effect for the retinal layer produced an F ratio of F (9, 630) = 51.64 and p < 0.0001. The interaction effect was also significant, F(54, 630) = 1.741, p = 0.0012. All algorithms classified with high accuracy (> 0.70) the innermost layers of the retina (total retina, inner retina, RNFL, GCL, INL) according to the gender of the participants, where we observed significant differences between genders in thickness and measurements volume. The SVC, LDA, and LR algorithms produced high accuracy (>0.70) when thickness and volume data came from the RNFL compared to the outermost layers of the retina. The KNN, RF and DT algorithms performed better in correctly classifying the total retina data in relation to the outermost layers. The thickness and volume of the retina and the innermost layers of the retina allow machine learning algorithms to be more accurate in separating data from different sexes.
Acesso aberto (Open Access)
Envelhecimento, declínio cognitivo e plasticidade astroglial em ca3
(Universidade Federal do Pará, 2011-11-18) TOKUHASHI, Tatyana Pereira; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
A few studies investigated in detail possible relationships between aging cognitive decline and hippocampal astroglial plasticity. In the present report we investigated in murine model possible relationships between performances in object recognition tests and the astrocytes laminar distribution in CA3. To do so, young (6 months old, n = 7) and old (20 months old, n= 5) C57Bl6 mice, were maintained in standard cages and assessed in object recognition hippocampal-dependent tasks. Isolated or integrated (episodic-like memory) tests were applied and revealed that object identity (What?), place (Where?) and time (When?), were impaired in old subjects, whereas in young mice only spatial memory was impaired. After behavioral tests all subjects were sacrificed and perfused with aldehyde fixatives had their brains removed and processed for glial fibrillary acid protein (GFAP) immunohistochemistry, a selective marker for astrocytes. To avoid sample bias we used the optical fractionator, a stereological method that is no affected by histological procedures. The results on behavioral isolated or integrated tests revealed that aging significantly impairs object, spatial and time recognition (two-tail t-test, p<0.05). As compared to young subjects, old mice showed laminar changes in the astrocytes distribution with proportional increase of the astrocytes number in the pyramidal layer of dorsal and ventral CA3 and a reduction in the lacunosum molecular layer of dorsal CA3. Coherently, the total number of CA3 astrocytes showed significant reorganization of its laminar distribution as a function of age with reduction of its numbers in the stratum oriens. No significant differences were detected in the mean values of laminar volumes suggesting that aging induced changes directly affected astroglial plasticity in CA3. Finally, a linear inverse correlation was found between the estimations of pyramidal cell layer astrocytes and performances in the behavioral tasks. Further direct evidences of this correlation with altered CA3 astrocytes and possible molecular mechanisms to explain aging cognitive decline remains to be investigated.
Acesso aberto (Open Access)
O fator de crescimento neuronal na infecção por Schistosoma mansoni: estudo molecular, imunoenzimático e morfométrico em modelo permissível e não permissível à infecção
(Universidade Federal do Pará, 2013-07-03) SANTOS, Daniel Valle Vasconcelos; SILVA FILHO, Manoel da; http://lattes.cnpq.br/2032152778116209
Schistosomiasis is a tropical disease caused by Schistosoma mansoni. His occurrence affects 110 million people worldwide. The deposition of eggs of the parasite may occur - in ectopic form – in the central nervous system (CNS) which leads to the formation of granulomas with consequent production of nerve growth factor (NGF). Since several studies have demonstrated the importance of NGF in the development of visual cortical pathways, our study aimed at evaluating the possible changes in the NGF concentratons in the visual system as well as the impact of this on the pyramidal cell morphology in two animal models. The change in concentration of the nerve growth factor as well as neuronal morphology were evaluated in suscetible and non-suscetible animals (mice and rats) to infection. We used 174 rats (Hooded Lister) and 135 albino mice bred and kept in cages and fed ad libitum. These animals were infected shortly after birth, with 50 cercariae. Seventy seven rats and 73 mice were inoculated with saline and constituted the control group of the study. The infection covered a period of 48 weeks . Samples of liver and visual cortex were removed, extracted and quantified with immunoassay kit (ChemiKineTM Nerve Growth Factor (NGF) Sandwich ELISA Kit - Chemicon International). For the morphometric analysis we used the pyramidal cells of the visual cortex layer IV marked by extracelular injection of biotinylated dextran (10,000 kDa). The results were expressed as mean ± standard deviation. We used Student t test to determine statistical differences between groups. The average value of NGF found in the visual cortex of rats infected was 39.2% higher than in the control group (infected: 400.9 ± 143.1 pg/ml, control: 288 ± 31.9 pg/mL, p < 0.0001). In liver samples, the increase was 28.9% higher in the infected group (infected: 340.9 ± 103.9 pg/mL, p < 0.01, control: 264.4 ± 38.6 pg/mL). No significant increase was detected within a week of infection. Among the mice group, the increase of NGF in the visual area was 94.1% (infected: 478.4 ± 284 pg/ml, p < 0.01; control: 246.5 ± 76.8 pg/ml). In the liver of these animals the increase was 138.7% (infected: 561.8 ± 260.7 pg/mL, p < 0.01, control: 301.3 ± 134.6 pg/mL). In mice group we found significant differences in dendritic parameters evaluated. The number of dendrites was 11.41% higher in the infected group than in the control (control: 25.28 ± 5.19; infected: 28.16 ± 7.45, p < 0.05). The total length of dendrites was also affected (control: 4916.52 ± 1492.65 μm; Infected: 5460.40 ± 1214.07 μm; p < 0.05), representing an increase of 11.06%. The total area of the dendritic receptive field was increased by 12.99% (control: 29.346,69 ± 11.298,62 μm2; Infected: 33.158,20 ± 7.758,31 μm2, p < 0.05) while the area had a somatic reduction of 13.61% (control: 119.38 ± 19.68 μm2; infected: 103.13 ± 24.69 μm2, p < 0.001). When we evaluated the effects of increased NGF in rats infected we did not observe significant differences in dendritic parameters analyzed, compared to the control group, except for an increase in the area of the neuronal body of approximately 21.18% (control: 132,20 ± 28.46 μm2; infected: 160.20 ± 31.63 μm2, p < 0.00001). This work showed that the reaction production of NGF in the CNS during infection with Schistosoma mansoni occurs in greater magnitude than permissible in the model in the model impermissible. We also demonstrated that in mice the effects on neuronal morphology is dramatically affected when the body is subjected to an increase in the concentration of NGF as a result of infection by Schistosoma mansoni. Given these data, studies evaluating the potential impact of visual effects and also in cell physiology caused by schistosomiasis infection becomes necessary to assess the actual damage caused by this pathological increase of nerve growth factor in the visual pathways of mammals.
Acesso aberto (Open Access)
Influência do tamanho da ninhada sobre o declínio cognitivo e a morfologia microglial da camada molecular do giro denteado em rattus novergicus
(Universidade Federal do Pará, 2012-10-11) OLIVEIRA, Marcus Augusto de; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286; DINIZ JUNIOR, José Antônio Picanço; http://lattes.cnpq.br/3850460442622655
It has been proposed that aging is associated with neuroinflammation in the central nervous system but it is not known whether microglial changes induced by aging are affected by early in life effects of litter size. On the other hand the molecular layer of dentate gyrus has been recognized as the main target of the perforant pathway, whose synaptic integrity is essential for the recognition memories of identity and spatial location. In the present report we investigated if aging cognitive decline and microglial morphological changes in the molecular layer are influenced by litter size changes early in life and aging. To assess these questions Wistar rats suckled in litters of six or 12 pups/mother were raised sedentarily in groups of 2-3 from the 21st post-natal day onwards. At four (mature adult) or 23 (aged) months of age were submitted to spatial memory and object identity recognition tests, sacrificed, perfused with aldehyde fixatives and had their brains processed for selective microglia/macrophages immunolabeling with anti-IBA-1 antibodies. A representative sample of the immunolabeled cells in the molecular layer of dentate gyrus was analyzed after three-dimensional reconstruction with Neurolucida software (Microbright Field Inc.) and morphological features of each cell were quantified by Neuroexplorer (Microbright Field Inc.). It was found that Wistar rats maintained all life in standard laboratory cages showed spatial memory deficits in both mature and aged subjects no matter the litter size. On the other hand all aged subjects independent of the litter size had their object recognition identity memory impaired. Microglial morphological analysis revealed that cell soma area and perimeter and branches volume seem to be more intensely affected by aging and that these changes are mainly associated with animals from large litters. In addition it was observed important shrinkage and thickening of the microglial branches in aged individuals in higher proportion in the group from large litters. Taken together the results suggest that spatial memory seems to be more susceptible to the aging process than object recognition and that these changes are associated with distinct effects on the soma and branching patterns of microglia of molecular layer from young and aged subjects.
Acesso aberto (Open Access)
Influências do ambiente e da idade sobre a complexidade morfológica dos astrócitos do giro denteado de camundongos suíços albinos
(Universidade Federal do Pará, 2015-05-14) FÔRO, César Augusto Raiol; SOSTHENES, Marcia Consentino Kronka; http://lattes.cnpq.br/7881527576747420; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
During our previous study (Diniz et al., 2010), mice (Mus musculus) maintained in an impoverished environment that mimicked a sedentary lifestyle from weaning generally performed worse on spatial memory tasks (the Morris water maze) and did not distinguish between old and recent or between displaced and stationary objects in episodic-like memory tests. In contrast, mice maintained in enriched cages for equal time preserved those abilities. These behavioral outcomes were associated with layer-dependent, numerical astrocytic changes. Using the same serial anatomical sections selectively immunolabeled for glial fibrillary acid protein from the previous study, we tested the hypothesis that environmental impoverishment would reduce the morphological complexity of astrocytes, and that such changes would be associated with learning and memory decline. We used three-dimensional microscopic reconstructions and unbiased systematic and random sampling approaches to select astrocytes from the polymorphic, granular, and molecular layers of the dentate gyrus. Cluster and discriminant analysis of three-dimensional astrocytic morphometric features from each layer and experimental group revealed two main morphological phenotypes. Type I astrocytes were more complex than type II; they exhibited larger tree areas, larger tree volumes, more segments, and more vertices. Integrated analysis with previous behavioral findings from the same animals revealed that the reductions in morphological complexity observed in young mice from impoverished and aged mice from enriched environments were observed in both astrocyte types in all layers of the dentate gyrus. We suggest that long-term environmental impoverishment and aging effects on astrocyte plasticity may represent at least part of the circuitry changes underlying learning and memory decline.
Acesso aberto (Open Access)
Influências do envelhecimento e do ambiente sobre a progressão da encefalite experimental por arbovírus Piry em modelo murino: mudanças morfológicas microgliais e alterações comportamentais
(Universidade Federal do Pará, 2014-10-03) SOUSA, Aline Andrade de; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
Environmental enrichment and aging effects on behavioral and microglial morphological changes were investigated in a murine model of sub-lethal arbovirus encephalitis. To that end two-months-old female albino Swiss mice were raised in impoverished (IE) or in enriched environments (EE) during 6 (young - Y) or 16 (aged - A) months. After behavioral tests, Y and A mice were nasally instilled with an equal volume of Piry virus infected (Py) or normal brain homogenates. Eight days post-infection (DPI), when behavioral tests first revealed sickness changes, mice brains sections were immunoreacted with anti-IBA1 and anti-Piry virus antibodies. At 20 and 40 dpi, the remaining infected animals were behaviorally re-tested, and processed for the same markers. In infected young mice from impoverished environment (IYPy), burrowing activity decreased and recovered earlier (8–10 dpi) than open field activity (20–40 dpi) but remained unaltered in age-matched mice from enriched environment (EYPy). In contrast, aged infected mice, both from enriched (EAPy) and impoverished (IAPy) environments, reduced significantly burrowing activity at all-time windows. Piry virus encephalitis induced transitory olfactory losses in IYPy and EYPy but permanent in IAPy and EAPy. Piry virus antigens immunolabeling reached a peak in CNS parenchyma at 5 and 6dpi and disappeared at 8dpi. All microglia three-dimensional reconstructions were done at 8dpi. Microglial changes were significantly more severe in young adult than in aged mice but EY mice seem to recover to the microglial homeostatic morphology earlier than IY. EE beneficial effects were smaller in aged mice.