Teses em Neurociências e Biologia Celular (Doutorado) - PPGNBC/ICB

URI Permanente para esta coleçãohttps://repositorio.ufpa.br/handle/2011/2390

O Doutorado 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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Análise transcriptômica das linhagens celulares B103 e C6 expostas à ação do metilmercúrio
(Universidade Federal do Pará, 2023-04) BONFIM, Laís Teixeira; FERREIRA, Wallax Augusto Silva Ferreira; OLIVEIRA, Edivaldo Herculano Correa de; http://lattes.cnpq.br/009400771470765; https://orcid.org/0000-0001-6315-3352
The intensification of anthropogenic activities produces a high rate of environmental pollution, mainly in water bodies, where the contamination by metals has become an object of great interest, due to their inability to support such load. Mercury (Hg) is a naturally occurring metal that can be used in the manufacture of home products such as fluorescent lamps, fungicides, and germicides. The entry of Hg into the food chain occurs through the methylation of Hg2+ ions into MeHg. After methylation, Hg is considered highly toxic to humans, and among the main target organs of this intoxication we can mention the brain, since MeHg easily crosses the blood-brain barrier and can accumulate in different brain areas. It is known that, once in the CNS, MeHg can cause extensive cellular damage, such as DNA damage, oxidative stress, neuroinflammation and cell death in both neurons and glial cells. Thus, the objective of this study was to analyze the transcriptomic alterations of cell lines B103 and C6, derived from neuroblastoma and glioma of Rattus norvegicus, exposed to the action of methylmercury. For this, the expression microarray technique was used to evaluate the global profile of gene expression after 24h of MeHg exposure. Our results demonstrate that MeHg induces significant alterations in gene expression of the two cell lines evaluated. The alterations were more prominent in the C6 cell line, in which a greater amount of differentially expressed genes was observed. Among the genes differentially expressed of the B103 cells we can highlight the genes Cdc42se2 (log2 FC -4.055713), Dcx (log2 FC 3.618981) and 4930449C09Rik (log2 FC 3.5129156) at a concentration of 0.1 μM. As for the exposure of 2.8 μM, the genes with the highest FC were Crem (log2 FC -4.027875), Otoa (log2 FC 3.501512) and Dcx (log2 FC 3.423433). In addition to the abovementioned genes, the genes Trim14, Gm14169, Gm30871, Otoa and Dcx were shared between the two exposed groups. As for the C6 lineage, ten transcripts with FC above 3 (Aldh1l2, Dac1, Rps4l, Zbtb46, 6430573p05Rik, Tcf12, Awat2, Muc3, Dclre1b, Slc38a6) are highlighted. In the 6.3 μM treatment, only three genes were altered more than 3 times (Rps4l, Ankdr44 and 2610318N02Rik). It is also noteworthy that three genes were shared between treatments (Rps4l, Lamb 3 and Gm 41386).
Acesso aberto (Open Access)
Avaliação do óleo essencial de pau-rosa (Aniba rosaeodora Ducke) em modelo de depressão induzida por álcool em ratas adolescentes
(Universidade Federal do Pará, 2023-08) SANTOS, Éverton Renan Quaresma dos; MAIA, José Guilherme Soares; http://lattes.cnpq.br/1034534634988402; MAIA, Cristiane do Socorro Ferraz; http://lattes.cnpq.br/4835820645258101
Depression is a prevalent disorder worldwide, which affects the functionality and quality of life of the people. The linalool-rich rosewood essential oil (Aniba rosaeodora Ducke) displays activities on the central nervous system (CNS), including of antidepressant-type. The work aimed to evaluate the effect of the essential oil in adolescent female rats through a model of binge drinking-induced depression. The essential oil was extracted by hydrodistillation and analyzed by a dual system of gas chromatography coupled to mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (CG-FID). The in vitro antioxidant capacity of the essential oil was determined in the 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging assay. Female Wistar rats, 29 days old, received distilled water or ethanol (3 g/kg/day) orally in 4 binge episodes, and saline solution or rosewood oil (35 mg/kg) intranasally once a day for 28 days. After the experimental treatment, splash and forced swimming tests were performed. The brain-derived neurotrophic factor (BDNF) and S100B gene expression, biological parameters of the oxidative stress and levels of pro-inflammatory cytokines were determined in the prefrontal cortex and hippocampus. The OEPR+EtOH group increased self-cleaning time and decreased immobility time, both significantly compared to the EtOH group in the splash and forced swimming tests, respectively. The essential oil behavioral effects were related to positive modulation of BDNF and S100B genes, to restoration of glutathione (GSH) and Trolox equivalent antioxidant capacity (TEAC) levels, in addition to the attenuation of the increase of thiobarbituric acid reactive species (TBARS), Interleukin-1β (IL-1β), Interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) induced by alcohol. The results suggest that essential oil improved the alcohol-induced depressive profile through a neuroprotective mechanism by positively modulating the gene expression of neurotrophic factors, rebalancing the antioxidant status and attenuating the inflammatory process, possibly due to the linalool action.
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A base molecular das adaptações visuais nos genes das opsinas de Anableps anableps e Phreatobius cisternarum através da análise de transcriptoma
(Universidade Federal do Pará, 2019-03-14) MARILUZ, Bertha Ruth Zelada; SCHNEIDER, Patrícia Neiva Coelho; http://lattes.cnpq.br/9584217233879031
Although vertebrate eyes share the same general organization, many species have developed specializations that improve their visual perception of the environment. These specializations are often reflected in a variety of visual adaptations that involve changes in visual sensitivity, which in turn can be modulated by the variation on the photoreceptors number, by altering the visual pigments or by combining both mechanisms. In the case of adaptive changes in the pigments, these may occur due to structural differences in the pattern of expression and the repertoire amount of the opsin genes family. However, there is no much information regarding adaptive changes on visual pigments over different luminosity on aquatic environments. This research aims to evaluate the molecular basis of the visual adaptations in the opsin genes from Anableps anableps and Phreatobius cisternarum, species present in environments with different light conditions, the first one from a surface environment and the second one from an underground habitat, through transcriptome analysis. This investigation comprises two chapters. The first chapter comprises the studies of the Anableps anableps specie. We combined the analysis of RNA-Seq and in situ hybridization of the eye tissue of this specie aiming to understand them visual adaptations to the aerial-aquatic environment. RNA-Seq data from the eye exhibited a repertoire of 20 non-visual opsin genes, which reflects the environmental heterogeneity these species lives. Likewise, comparative analyzes in protein coding sequences of the opsins allowed the identification of six opsins presenting the typical amino acid motifs of C-type and nine of Group 4, conserved among themselves. In situ hybridization studies on the retina showed asymmetric expression of these non-visual opsins at different stages, as well as during the ocular development. The second chapter presents the studies of the Phreatobius cisternarum specie. We combine histological, molecular and RNA-Seq analyzes to understand the visual and sensorial adaptations of P. cisternarum to the phreatic environment. RNA-Seq data from P. cisternarum head revealed a repertoire of eleven opsin genes, three visual opsins and eight non-visual opsins. Two visual opsins, rh1 and lws, presented amino acid substitutions that potentially contributed to the red and blue shift, respectively. Our histological analysis showed the presence of a rudimentary retina, while the RNA-Seq analysis identified the expression of 38 genes related to the lens fiber cells and 51 genes related to the retinal pigmented epithelium (RPE), indicating that the reduced eyes of P. cisternarum retained some lens structures. The extra-retinal expression of opn4m3 is possibly associated with the peripheral clock regulation. Furthermore, the presence of potential opsin pseudogenes would be regulated by a small retina exposed to a low-light environment. The chapters introduce and provide an overview of the investigation on opsins amino acid substitutions, changes on expression patterns and opsin repertoire size (duplication and pseudogenization), and how these could contribute to the shift in spectral sensitivity and finally visual adaptation of A. anableps and P. cisternarum to their peculiar environment. The present study provides fundamental evidence for the understanding of the adaptive molecular basis in the opsin genes to subterranean and aerial-aquatic environments, in the species P. cisternarum and A. anableps.
Acesso aberto (Open Access)
Efeitos do exercício físico sore parâmetros cognitivos e bioquímicos em ratos expostos ao etanol de forma intensa e episódica (Binge Drinking)
(Universidade Federal do Pará, 2018-12-21) TEMBRA, Dinair Pamplona dos Santos; LIMA, Rafael Rodrigues; http://lattes.cnpq.br/3512648574555468
The consumption pattern of heavy and episodic ethanol, weekend consumption, characterizes the pattern of excessive alcohol consumption or binge drinking that promotes an imbalance of brain metabolic functions, contributing to neurodegeneration and cerebral dysfunction. And because it is a legal drug, it has global relevance in public and social health. In this way, we aimed to investigate the effects of physical training of moderate intensity, in treadmill, on the deleterious effects of ethanol on hippocampus functions related to memory and learning. For this, 80 Wistar rats were divided into four groups: Control group; Trained group (animals trained and treated with distilled water); Ethanol group (animals not trained and treated with doses of 3 g / kg / day of ethanol, 20% w / v); and ethanol + trained group (animals trained and exposed to ethanol). Physical exercise was performed on a treadmill for 5 days a week for 4 weeks and all doses of ethanol and distilled water were administered by intragastric gavage (three days a week) in four repeated cycles. After the experimental period, the animals were submitted to the task of object recognition and Morris aquatic labyrinth test, and after euthanasia, blood and hippocampus were collected to measure levels of antioxidant capacity equivalent to trolox (TEAC), content of reduced glutathione (GSH), nitrite and lipid peroxidation. (LPO). Our results showed that EtOH caused marked oxidative stress and mnemonic damage, and physical exercise promoted neuroprotective effects, including modulation of oxidative plasma biochemistry (by restoration of GSH levels) and hippocampus (reducing levels of LPO and increasing antioxidant parameters) and improving cognitive function. Therefore, physical exercise may be an important prophylactic and therapeutic tool to improve and even prevent the deleterious effects of ethanol on cognitive functions.
Acesso aberto (Open Access)
Estudo eletrorretinográfico da adaptação à luz de vias de processamento específicas de cone e de oponência em cor e luminância
(Universidade Federal do Pará, 2017-12-15) COSTA, Alódia Brasil; SILVA, Anderson Manoel Herculano Oliveira da; http://lattes.cnpq.br/8407177208423247
Visual system has an essential function called light adaptation or photopic adaptation, which consists in the regulation of light sensitivity allowing visual adaptation to a wide range of illumination levels. This phenomenon is not fully elucidated. Therefore, the present study aims to evaluate the magnitude and temporal pattern of the variation of retinal electrical responses for L cone and M cone selective stimuli and for luminance pathway and red-green color pathway specific stimuli, making possible the evaluation of the red-green parvocellular (P pathway) and magnocellular (M pathway) visual pathways activity, during the photopic adaptation. For such purpose, 6 healthy subjects underwent 30 minutes of dark adaptation and then were exposed a light steady background for 16 min. The stimuli were given every 2 min and the electrical retinal response was recorded by full field electroretinogram (ffERG) over 16 min. The triple silent substitution method was used to isolate the responses of pathways originating from the L or M cone. We also used specific stimuli of luminance (Lum) and red-green color (Crom). For each stimulation type was used intermediate (12 Hz, which reflects the activity of the red-green parvocellular pathway - P pathway) and high (36 Hz, which reflects the activity of the magnocellular pathway - M pathway) temporal frequency, resulting in 8 stimulation conditions. Amplitude and phase of the first (F), second (2F) and third (3F) harmonic components were extracted by Fast Fourier Transform. It was observed that amplitude and phase increased over light adaptation time with simple sine wave form in the most of components and stimulation conditions. The relative increases in F amplitude from the M-cone driven responses during the light adaptation were higher than those L-cone driven ERGs at both temporal frequencies, 12 Hz (M = 1.21, L = 0.33) and 36 Hz ( M = 1.94, L = 0.55), as well as they were higher at 36 Hz than at 12 Hz for the two cones. In general, there was slight F phase increase during the light adaptation time (<30 degrees), slightly higher at 36 Hz. Regarding the light adaptation kinetics, F amplitude and phase that seem reflect the activity of the P pathway presented a faster adaptation (L-cone 12 Hz, M-cone 12 Hz, Crom 12 Hz, Lum 12, with a mean of 1.4 min) and those that probably reflect activity of the M-pathway had slower adaptation (Lum 36 Hz, L-cone 36 Hz, M-cone 36 Hz, with a mean of 4.9 min). Thefore, M and P pathways presented different magnitude and kinetics of light adaptation, being the M pathway the pathway of greatest increase and slower adaptation.
Acesso aberto (Open Access)
Investigação de tipos e origem celular neurogênicos em áreas diversas do sistema nervoso central da espécie Cebus apella
(Universidade Federal do Pará, 2015-07-27) SANTOS, Adriano Guimarães; HAMOY, Moisés; http://lattes.cnpq.br/4523340329253911; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
Identifying populations of neuronal precursors and stem cells generated by own central nervous system has held discussions on its possible use in the repair of damage caused by acute disorders of the central nervous system. However, in the case of adult mammalian brain, such generation is considered evolutionarily restricted to two areas: subgranular zone of the dentate gyrus of the hippocampus, and the walls of the lateral ventricles where new neurons are continuously generated. We used 6 non-human primates, adults, males of the species Cebus (capuchin monkey, 10 years old), weighing between 2.1 and 2.8 kg (mean 2.5 kg) pretreated with BrdU that after sacrifice and because histological processing, their tissues were analyzed for immunohistochemical analysis using anti-BrdU anti-Nestin, and Sox2 anti-DCX in diverse areas of the Central Nervous System. The ventricular walls showed the presence of neuroblasts similar to that observed in previous studies, but unexpected results were also observed in areas such as the frontal cortex.
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Isolamento e cultivo in vitro do agente etiológico da Doença de Jorge Lobo: morfologia, fisiologia e genoma de Candida loboi sp. nov
(Universidade Federal do Pará, 2015-12-18) COSTA, Patricia Fagundes da; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Jorge Lobo’s disease is a chronic granulomatous infection developing after traumatic implantation of the fungus in the skin. It presents with nodular, verrucous or keloid-like lesions mainly on legs and ears. The highest prevalence is at Amazon Region and it has been considered an emergent disease, presenting new cases on other continents in both, humans and dolphins. Little is known about L. loboi, and the absence of in vitro culture impairs the correct characterization of the fungus. This work had as the main goal to isolate, culture and characterize strains of the etiological agent of Jorge Lobo´s disease, obtained from patients attended at the Dr Marcello Candia Reference Unit in Sanitary Dermatology of the State of Pará, in Marituba, Pará, Brazil. During many years 23 patients were diagnosed and followed by our team. Most of them were male farmers, with age varying from 14 to 80 years-old. After biopsy of the lesion for confirming the diagnosis, the patients initiated treatment and the material was processed with dispase II at liquid medium RPMI, 37º C, 5% CO2 for 1-2 weeks to isolate yeast cells from human tissue. After 2-6 months in the medium, we observed fragmentation of mother cells and the presence of new yeast cells with diameter varying from 1 to 7μm. From that moment, it was possible to grow the strains in different liquid or solid mediums at 37º C or RT, where creamy, whitish cerebriform, sometimes hairy colonies were observed. Cells from those strains were analyzed by different techniques of optical and electron microscopy, biochemistry and genetics, ending with the description of the genome of one patient just after isolation of the fungal cells from the lesional skin, defining the following taxonomic identification: Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes; Saccharomycetales; Debaryomycetaceae; Candida/Lodderomyces clade; Candida; Candida sp. LDI48194. The peculiar clinical presentation associated to unique morphological, physiological and genetic characteristics that do not permit the definition of a known species indicate that the etiological agent of Jorge Lobo’s disease is a new species, for which we propose to use the name Candida loboi.
Acesso aberto (Open Access)
O que os olhos não vêem o coração não sente: implicações para a opinião sobre a redução da maioridade penal
(Universidade Federal do Pará, 2018-04-27) CALDAS, Ivete Furtado Ribeiro; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170
The increase in urban violence has led society to charge the State with more severe and punitive measures to solve the problem of juvenile crime. One of the proposals is the reduction of the criminal majority from 18 to 16 years. Discussions on this proposal have been ideologically polarized and instances where the debate is based on reliable scientific evidence are rare. In this research, we try to contribute to identify the implicit social and moral constraints associated with the issue of criminal majority. For this, we designed two experiments to evaluate how socioeconomic factors, sociodemographic factors and the moral development of the individual influence the opinion on the topic. Objective: To verify the relationship between socioeconomic factors, sociodemographic factors and moral development in the opinion about the reduction of the penal majority. Materials and Methods: This is a qualitative and quantitative research with a cross-sectional design that consisted of two experiments: Belem Experiment and Regional Experiment. In the Belém Experiment the data were collected in two public places of the municipality of Belém, Pará, Praça Batista Campos and the Court of Justice of the State of Pará. In the Regional Experiment an on line tool was used with the aim of achieving people from different regions of the Brazilian territory. The sample consisted of individuals older than 18 years of both sex. The instruments used were: (1) Socioeconomic and sociodemographic questionnaire and (2) Dilemma of the adolescent in conflict with the law. Pearson's Chi-square test (χ2) and data mining were used for data analysis. Results: Juvenile court workers display preference for lower stages (stage 1) and a lower level of moral competence (average c-index) (3.97), with a strong adherence to the law, different from the general public that prefers higher stages (stage 6) and c-index (14.29). There was a significant relationship between the stage preference and the opinion of the subjects, only in the juvenile court workers (χ2 = 20,665, df = 10, p = 0.024). However, the former is less in accordance with lowering the age of criminal responsibility than the latter. Conclusion: The accuracy of opinion is greater at the extremes of psychological distance, that is, when the individual is very distant or very close to the reality of this adolescent.

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