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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Acesso aberto (Open Access)
Acidente vascular encefálico isquêmico na exposição crônica ao etanol: estudo pré-clínico da comorbidade e da resposta a minociclina
(Universidade Federal do Pará, 2015-02-27) FONTES JÚNIOR, Enéas de Andrade; MAIA, Cristiane do Socorro Ferraz; http://lattes.cnpq.br/4835820645258101; CRESPO LÓPEZ, Maria Elena; http://lattes.cnpq.br/9900144256348265
Stroke is the second largest cause of death in the world and the leading in Brazil, with 87% of strokes due to ischemic processes. Chronic ethanol consumption, usually beginning in adolescence, is recognized as an independent risk factor for increased morbidity and mortality by stroke. Although cases combining the two diseases are relatively common, there is no data in animals or clinical models demonstrating the quality or mechanisms of interaction between the two morbidities, nor its impact on therapeutic intervention. Considering the recent studies proposing minocycline as a new therapeutic tool for the treatment of stroke, this study aimed to investigate the interaction between the Chronic Alcoholic Intoxication (CAI) started in adolescence and the stroke in motor cortex of adult rats, and the effects of treatment with minocycline on this interaction, using behavioral, cellular and molecular parameters. Female Wistar rats (35 days-old) were chronically exposed to ethanol (6.5 g/kg/day, 22.5% w/v) or water for 55 days. One day after the end of the CAI focal ischemia was induced in motor cortex with the endothelin-1 (ET-1), followed by seven-day treatment with minocycline or saline. After this period, the animals were assayed with open field and rota rod tests. Immediately, animals were sacrificed and cortex was dissected for evaluation of nitrite and lipid peroxidation levels. In all groups, some animals were perfused and the motor cortex subjected to histological analysis to assess the damage, and immunohistochemical labeling to neuronal death (anti-NeuN), microglial/macrophage (anti-ED1) and astrocytes (anti-GFAP) activation. The ethanol intoxication from puberty to adulthood potentiated the damage caused by stroke, causing major losses in capacity to start and running movements as well as the strength and motor coordination compared to ischemic animals pretreated with water. These manifestations were accompanied by increased neuronal loss, reduced ED-1+ and GFAP+ cells and higher levels of nitrite and lipid peroxidation. Treatment with minocycline was effective in preventing/reverse motor deficits and tissue damage induced by focal ischemia, also inhibiting the increase in oxidative stress markers. The CAI either alone with succeeded by focal ischemia, harmed the outcome of treatment with minocycline. Our results indicate that heavy alcohol intoxication during adolescence exacerbates the motor deficit and tissue damage in animals subjected to focal ischemia. This process appears to be associated with microglia/astroglial activation, but mainly with oxidative stress. It also shows that the previous history of CAI started adolescence interferes significantly in the treatment of cerebral ischemia with minocycline.
Acesso aberto (Open Access)
Ajustes motores compensatórios após lesão isquêmica focal unilateral do trato corticoespinhal
(Universidade Federal do Pará, 2017-06-30) CARVALHO, Walther Augusto de; BAHIA, Carlomagno Pacheco; http://lattes.cnpq.br/0910507988777644; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170
The aim of this work was to develop a new model of spinal cord injury caused by focal and unilateral transient ischemia after ET-1 microinjection in the dorsal funiculus and to evaluate the sensorimotor alterations of the anterior paw of rats (Wistar). Fifty (n = 50) animals (CEPAE / UFPA protocol BIO007912), who were trained, thirty-three (n = 33) were selected to compose control (n = 15), sham (n = 6) and injury (n = 12) groups. By using a micropipette, we injected the volume of 250 nL of saline (sham) or endothelin-1 (lesion) near the medial dorsal artery of the cervical segment C4 at a depth of 1 mm from the pial surface of the spinal cord. ET-1 induced cystic cavity formation of 0.421 mm2 (± 0.035 mm2, n = 3) on the corticospinal tract and suprajacent white matter, ipsilateral to the microinjection site that can be measured in cross-sections (50 μm) stained by the Nissl technique. The motor functions of the forepaw were evaluated by specific sensorimotor tests before and after injury at 3, 7 and 14 days. The results were evaluated by the ANOVA statistical test with Tukey post-hoc analysis (α = 0.05). Our results show in pasta test that after injury there is a compensatory motor behavior in which the non-preferential forepaw assumes the functions of the preferential forepaw. The Staircase test revealed a decrease in the ability to grasp the object with the preferred paw and the Contact test showed a decrease in sensitivity of the preferred paw.
Acesso aberto (Open Access)
Ajustes posturais antecipatórios e compensatórios em idosos com e sem lombalgia
(Universidade Federal do Pará, 2021-03) GARCEZ, Daniela Rosa; CALLEGARI, Bianca; http://lattes.cnpq.br/0881363487176703; https://orcid.org/0000-0001-9151-3896; YAMADA, Elizabeth Sumi; http://lattes.cnpq.br/7240314827308306
Chronic low back pain (DLC) is associated with changes in postural control and is highly prevalent in the elderly. Research shows that aging and DLC are described as important factors that affect postural control. The postural control impairments increase the risk of falls. Researches evaluating the postural control in elderly people with DLC are still necessary for greater effectiveness in balance rehabilitation programs to prevention falls in this population. The objective of this study is to verify whether anticipatory postural adjustments (APAs) and compensatory postural adjustments (CPAs) are affected by DLC in elderly people by assessing their postural control during a self-initiated perturbation paradigm induced by rapid upper arm movement when pointing to a target. Elderly people were divided into: Group with DLC (GDLC) (n = 15) and Control Group (CG) (n = 15). The participants’ lower limb muscle onset and center of pressure (COP) displacements were assessed prior to perturbation and throughout the entire movement. T0 moment (i.e., the beginning of the movement) was defined as the anterior deltoid (DEL) onset, and all parameters were calculated with respect to it. The rectus femoris (RT), semitendinosus (ST), and soleous (SOL) showed delayed onset in the GDLC group compared with the control group: RF (control: -0.094 ± 0.017 s; GDLC: -0.026 ± 0.012 s, t = 12, p < 0.0001); ST (control: - 0.093 ± 0.013 s; GDL: -0.018 ± 0.019 s, t = 12, p < 0.0001); and SOL (control: -0.086± 0.018 s; GDL: -0.029 ± 0.015 s, t = 8.98, p < 0.0001). In addition, COP displacement was delayed in the GDLC group (control: -0.035 ± 0.021 s; GDL: -0.015 ± 0.009 s, t = 3; p = 0.003) and presented a smaller amplitude during APA COPAPA [control: 0.444 cm (0.187; 0.648); GDLC: 0.228 cm (0.096; 0.310), U = 53, p = 0.012]. The GDLC group required a longer time to reach the maximum displacement after the perturbation (control: 0.211 ± 0.047 s; GDLC 0.296 ± 0.078 s, t = 3.582, p = 0.0013). This indicates that GDLC elderly patients have impairments to recover their postural control and less efficient anticipatory adjustments during the compensatory phase. Our results suggest that people with GDL have altered feedforward hip and ankle muscle control, as shown from the SOL, ST, and RT muscle onset. This study is the first study in the field of aging that investigates the postural adjustments of an elderly population with GDLC. Clinical assessment of this population should consider postural stability as part of a rehabilitation program.
Acesso aberto (Open Access)
Alteração diferencial nos astrócitos radiais do hipocampo e neurogênese em aves marinhas com rotas migratórias constantes
(Universidade Federal do Pará, 2019-08-17) LIMA, Camila Mendes de; MAGALHÃES, Nara Gyzely de Morais; http://lattes.cnpq.br/2519507561210918; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286; https://orcid.org/0000-0001-6611-6880
Little is known about environmental influences on radial glia–like α cells (radial astrocytes) and their relation to neurogenesis. Because radial glia is involved in adult neurogenesis and astrogenesis, we investigated this association in two migratory shorebird species that complete their autumnal migration using contrasting strategies. Before their flights to South America, the birds stop over at the Bay of Fundy in Canada. From there, the semipalmated sandpiper (Calidris pusilla) crosses the Atlantic Ocean in a non-stop 5-day flight, whereas the semipalmated plover (Charadrius semipalmatus) flies primarily overland with stopovers for rest and feeding. Using hierarchical cluster and discriminant analysis of morphometric features to classify three-dimensionally (3D) reconstructed cells, we identified two morphotypes of radial glia, designated as Type I and Type II. The migratory process affected these cells differentially, with more intense morphological changes in Type I than in Type II morphotypes in both species. We also compared the number of doublecortin (DCX)-immunolabeled neurons with morphometric features of radial glial–like α cells in the hippocampal V region between C. pusilla and C. semipalmatus before and after autumn migration. Compared with migrating birds, the convex hull surface of radial glial–like α cells of wintering birds significantly increased in both C. semipalmatus and C. pusilla. This increase correlated with an increase of the total number of DCX-immunolabeled neurons in wintering birds. The decreased radial astrocyte morphological complexity in the semipalmated sandpiper and its increase in the semipalmated plover, a species that probably relies more on visuospatial information for navigation, may be significant, despite phylogenetic and other differences between these taxa. The migratory flight of the semipalmated plover, with stopovers for feeding and rest, versus the non-stop flight of the semipalmated sandpiper may differentially affect radial astrocyte morphology and neurogenesis.
Acesso aberto (Open Access)
Alterações da formação hipocampal do Calidris pusilla associadas à migração outonal de longa distância
(Universidade Federal do Pará, 2017-08-31) MAGALHÃES, Nara Gyzely de Morais; DINIZ, Cristovam Guerreiro; http://lattes.cnpq.br/1025250990755299; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
After breeding in the upper Arctic tundra, shorebirds affected by migratory restlessness trace an inherited preliminary route and use compasses, maps and visual landmarks, until they reach, in the northern hemisphere, stopover sites that have the necessary nutritional resources for fast and high gain of energy reserves for migratory journey, as in the Bay of Fundy-Canada. Following this stopover site that is used by 75% of the population of Calidris pusilla, the long-distance autumn migratory experience continues with uninterrupted 6-day non-stop flights over the Atlantic until these birds reach South America and then the island of Canela-Brazil. To test the hypothesis that the long-distance migratory process would influence neurogenesis, astrogenesis and activation of earlier-expression genes, we captured 12 individuals in full migratory activity in the Bay of Fundy and 9 individuals in the Island of Canela in Brazil. After selective immunostaining for mature neurons (NeuN), immature neurons (Dcx), astrocytes (GFAP), and neuronal activation by early genes (c-Fos), we quantified these markers in the hippocampal formation and compared the results of this quantification of the individuals in migration (Bay of Fundy) with those of wintering birds (Canela Island). We used quantitative stereological analyzes to estimate the total number of cells of hippocampal formation, number of active cells, total number of astrocytes and young and mature neurons. To verify if the differences found were statistically significant, we used the Student t test. Our results confirmed that autumnal migration alone, caused hippocampal changes in Calidris pusilla. After migration, we detected that the hippocampal formation has fewer activated cells and fewer astrocytes, more new neurons and greater relative volume in the quantified hemisphere (left hemisphere). However, this process did not influence the number of total cells and mature neurons. We suggest that the difference found between the volume and number of new neurons, of the full migration and wintering individuals, possibly occurred due to the migratory process in combination with local conditions found during the beginning of the wintering period. Taken together our findings demonstrate long-distance migratory shorebirds offer a unique opportunity to investigate many issues related to the cellular neurobiology of migration in general, and, on the neural plasticity associated with hippocampal neuronal and neurogenesis in adult birds.
Acesso aberto (Open Access)
Alterações genéticas e epigenéticas em meningiomas na população paraense
(Universidade Federal do Pará, 2013-07-17) BASTOS, Carlos Eduardo Matos Carvalho; ANSELMO, Nilson Praia; http://lattes.cnpq.br/6518287721873199; NAGAMACHI, Cleusa Yoshiko; http://lattes.cnpq.br/8887641213110093
Meningiomas are the most common intracranial tumors that originate from the meninges surrounding the brain and spinal cord. Despite meningiomas were among the first solid neoplasms to be studied cytogenetically, little is known about their genetic and epigenetic profile. This study aimed to investigate genetic and epigenetic alterations that could contribute to tumor initiation and progression in meningiomas in the population of Pará, Brazil. This thesis is subdivided into three chapters. In Chapter I we investigated the association between the MTHFR C677T and meningioma in 23 patients in the population of Pará. A total of 96 healthy individuals with no previous pre-neoplastic lesions were selected for the control group. This association was not found. Although not statistically significant, our observation suggests that the TT genotype increases the risk of developing meningioma when compared to CC genotype. In Chapter II we evaluated the methylation pattern in two members of microRNA124 family in meningiomas in the population of Pará. Hypermethylation of the promoter region of miRN124a2 and miRNA124a3 appears to be a frequent event, as was found in 73.9% and 69.56% of the samples, respectively. In Chapter III, we analyzed the methylation pattern of the APC, BRCA1, CDH1, CDH13, CDKN2A, DAPK1, ESR1, FHIT, GSTP1, MGMT, MLH1, NEUROG1, PDLIM4, PTEN, Rb, RASSF1, RUNX3, SOCS1, TIMP3, TP73, VHL and WIF1 genes in a grade I and in a grade II meningiomas through an assay developed by MethylScreen. Pattern of methylation of CDKN2B was also analyzed in 25 patients with meningioma through bisulfite conversion, PCR and direct sequencing. RASSF1A was methylated in 16.73% and 63.66% of the CpG sites analyzed in the grade I and grade II meningioma, respectively. RUNX3 is methylated only in grade II meningioma in 52.88% of the CpG sites analyzed. Our results point to the importance of epigenetic changes in tumorigenesis and tumor progression in meningiomas.
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)
Alterações histopatológicas dos rins de macacos prego, Cebusapella (Linnaeus 1758) após exposição crônica a baixas doses de metilmercúrio
(Universidade Federal do Pará, 2014-02-28) SOUSA, Andréa do Socorro Campos de Araújo; SILVEIRA, Luiz Carlos de Lima; http://lattes.cnpq.br/9383834641490219
Mercury has been a major environmental and occupational risk and it still remains a problem for human health in the Amazon region. Although studies have shown that mercury affects various tissues and organs, kidneys are the target organs to the metal toxicity. Thus, the aim of this study was to investigate the effects of chronic exposure to low doses of methylmercury on renal parenchyma of Cebusapella, adult males exposed during 120 consecutive days with daily oral doses of 1.5 μg in the diet. The concentrations of total mercury in the animals’ blood were monitored every 30 days using a cold vapor atomic spectrophotometer (201 Hg), compared to the control group. The method used for histopathological analysis was the immersion in paraffin for staining with hematoxylin and Eosin, Masson's CAB and PAS. The immunohistochemical investigations included reactions for detection of smooth muscle actin ( IA4 ), muscle actin ( HHF35 ) and cytokeratin (AE1 and AE2). The results showed that treatment with mercury caused significant differences (P < 0.001) between the exposed and control groups. As for total Hg levels, histopathologicalchanges just likehydrops in Proximal tubuleswere observed, a common finding in methylmercury exposure in other species, with no significant changes in creatinine and urea concentrations. The Person correlation test showed a strong negative relationship between mercury concentration and animal body weight loss (P < 0.0001). Another important finding was the decrease in mesangial cells number, which suggests that methylmercury executed its nephrotoxicity, affecting not only the renal tubular system, as well as the glomerular mesangium cells, making necessary a greater amount of experimental studies to clarify which mercury concentration alert level is capable of triggering aggression mechanisms and kidney injury in subjects exposed to methylmercury.
Acesso aberto (Open Access)
Alterações mitocondriais e tumorigênese de câncer gástrico em Sapajus apella
(Universidade Federal do Pará, 2018-06-15) ANTUNES, Symara Rodrigues; BORGES, Bárbara do Nascimento; http://lattes.cnpq.br/0676220027193876
Cancer is the name given to a variety of diseases that can occur in different regions of the body, which is characterized primarily by the deregulated proliferation of cells. A very important organelle in both normal and mutated cells is mitochondria, responsible for most of the ATP production in the cell. Mutations in mitochondrial DNA can lead to apoptosis or influence the efficiency of ATP formation. Considering several different estimates, gastric cancer still in the five most incidental in world population, as well as in Brazilian and local population. In this way, understanding tumor behavior becomes important for fight against this pathology. With this, the objective of the present work was to analyze presence of mitochondrial DNA alterations of gastric carcinoma lines implanted in an animal model. Four mitochondrial genes (COI, ATPase 8, ND1 and ND3) from four gastric cancer strains (AGP01, ACP02, ACP03 and PG100) and one control (Carcinossarcoma 256 from Walker) were analyzed to evaluate possible mitochondrial DNA mutations. These strains were inoculated in non-human primates of the Sapajus apella species, and some animals received the carcinogenic substance N-methyl-N-nitrosurea (MNU) concomitantly with the strains. The gastric tumors that developed in the animals were surgically removed, after which DNA extraction, amplification and sequencing of the sequences of interest were done. Changes were observed in the ND1 and ND3 genes. The two transitions found in ND1, one at position 3594 (CT) and 3693 (GA) of mitochondrial DNA, had no associated pathological record and were related to population markers. The AG transition at position 10398 of the ND3 gene resulted in the change from one threonine to alanine in the resulting amino acid, only in lines with more aggressive behavior or after MNU administration. Two heteroplasms were also identified in the ND1 gene at positions 3594 (C / T) and 3693 (A / G) only in the PG100 line after MNU, suggesting a difference in the DNA repair system of this line compared to the others. The results suggest that changes in the genes encoding proteins that participate in Complex I of the respiratory chain are more frequent than in other portions of the mtDNA in the analyzed gastric carcinoma strains.
Acesso aberto (Open Access)
Alterações neuroquímicas no tecido retiniano murino em modelo de malária cerebral induzida pela infecção por Plasmodium berghei (ANKA)
(Universidade Federal do Pará, 2011-07-21) OLIVEIRA, Karen Renata Matos; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978
Cerebral Malaria (CM) is a severe complication resulting from Plasmodium falciparum infection. This condition has usually been associated with cognitive, behavioral and motor dysfunctions, being the retinopathy the most serious consequence resulting from the disease. The pathophysiologymechanisms underlying the complications of CM remain incompletely understood. Several experimental models of CM have already been developed in order to clarify those mechanisms related to this syndrome. In this context, the present work has been performed to investigate which possible neurochemistry alteration could be involved in the CM pathology. Male and female susceptible C57Bl/6 mice (6-8 week old) infected with ≈106 parasitized red blood cells (PbA), showed a low parasitaemia (15-20%), with evident clinical signs as: respiratory failure, ataxia, hemiplegia, and coma followed by animal death. In parallel to the clinical characterization of CM, retinal analysis demonstrated that the disease led to a decrease in the glutathione levels with 2 days post inoculation. However, this decrease was not so evident with the course of the infection (4º and 6º days post- infection). We further demonstrated that the increase in the glutathione levels during the infection is followed by the increase in the 3H-glutamate uptake rate (4º and 6º days post-infection), suggesting that CM condition causes an up-regulation of the transporters systems. Immunofluorescence data demonstrated that besides the activity increases, CM condition also stimulated the increase of the xCG- system expression in the retinal tissue. Furthermore, our findings also highlighted that in the retina the neurochemistries alterations occurs in a manner independent on the establishment of an inflammatory response, once TNF-α levels and NOS-2 expression were altered only in the cerebral tissue.
Acesso aberto (Open Access)
Alterações oxidativas e inflamatórias induzidas pela dapsona no sangue e no córtex pré-frontal de camundongos: efeitos do ácido alfa-lipóico
(Universidade Federal do Pará, 2018-12-14) GOMES, Bruno Alexandre Quadros; MONTEIRO, Marta Chagas; http://lattes.cnpq.br/6710783324317390
Dapsone (DDS), a drug used in leprosy multidrug therapy, can cause many adverse reactions and intoxications, inducing the generation of reactive oxygen species (ROS) and imbalance in the redox state, increase methemoglobin (MetHb) formation, hemolysis and release of heme and iron free, which may interfere with redox homeostasis in more vulnerable regions, such as prefrontal cortex (PFC), causing neurotoxicity and even neuroinflammation. In this sense, antioxidant compounds with chelating properties such as α-lipoic acid (ALA) may play a key role in combating or preventing these alterations. Thus, this work aims to evaluate the effect of DDS on MetHb formation, peripheral oxidative stress, and oxidative changes and neuroinflammation in PFC, as well as, effects of ALA. For this, was induced MetHb formation in Swiss mice with DDS 40mg/kg ip for 5 days. Two hours after DDS administration, ALA was given at two concentrations (12.5 and 25 mg/kg). Besides MetHb percentage, total equivalent antioxidant capacity (TEAC), reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) thiobarbituric acid reactive substances (TBARS), and iron concentrations in blood and PFC were evaluated, as well as, IL-1β, IL-17, and IL-4 cytokine concentrations, and de F4/80+, GFAP, and BDNF expression in PFC. Our results show that DDS induces the MetHb formation in red blood cells of mice, however, ALA was able to prevent or reverse the oxidation of hemoglobin induced by DDS at two used concentrationns. DDS reduced antioxidant capacity (TEAC) in plasma and red blood cells; decreased erythrocyte GSH, CAT, and SOD; and increased TBARS and plasma iron; however, ALA at two concentrations increased or reestablished TEAC in plasma and red blood cells at baseline levels. In addition to increasing or reestablishing GSH levels, SOD, and CAT in red blood cells, and decreased TBARS and iron levels, mainly in euthanized animals 4h after treatment. Curiously ALA 50mg/kg increased plasma iron concentrations. The treatment with DDS 40mg/kg also reduced TEAC, GSH, SOD e CAT in the PFC of the mice and increased TBARS and iron, characterizing oxidative stress, mainly in euthanized animals in 24h after treatment. Treatment with ALA increased or restored TEAC and GSH; and increased SOD and CAT in 12,5mg/kg concentration in euthanized animals 4h after treatment, as well as reducing TBARS levels and decreasing or preventing iron overload, mainly in euthanized animals 24h after treatment. DDS also promoting microglial and astrocyte activation in PFC, through F4/80+ e GFAP expression., with increased IL-1β and IL-4 production, and BDNF reduction, on the other hand, ALA 25mg/kg reduced GFAP and IL-1β expression, besides increased BDNF, suggesting that DDS also can cause neuroinflammation, and ALA presents antioxidant and anti-inflammatory properties against toxicity caused by DDS. These results suggest that ALA is promising and plays an important role in the prevention and/or formation of MetHb, reestablishment of redox balance and iron concentrations in both blood and PFC. Thus, ALA may be a usefull adjuvant therapy in DDS-induced toxicity, with lower toxicity and increasing adherence to treatment of leprosy patients.
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 de parâmetros de exposição mercurial, suscetibilidade genética e intoxicação em populações ribeirinhas do Tapajós e Tucuruí
(Universidade Federal do Pará, 2016-08-30) ARRIFANO, Gabriela de Paula Fonseca; CRESPO LÓPEZ, Maria Elena; http://lattes.cnpq.br/9900144256348265
Mercury is a heavy metal responsible for intoxications worldwide. Most toxic form is methylmercury that has affinity for the central nervous system, with recognized neurotoxicity. Some regions of the Amazon are well characterized by mercury exposure in humans, as the region of the Tapajos, due to local mining activity, for example. However, others, such as Tucuruí, remain virtually unstudied, with only one study in humans to date. In the Amazon, there is a large number of studies showing mercury exposure, however, intoxication and susceptibility studies are far less numerous in the Amazonian populations, and even today, there is no study analyzing simultaneously the three factors. The objective of this study was to determine the exposure (mercury content in the body by using mercury levels in hair samples), the individual susceptibility (genetic predisposition of each individual to suffer a damage because the exposure, using the genotyping of apolipoprotein E) and the intoxication (quantifying the extent of the damage already caused by using biomarkers such as S100B and NSE) in Amazonian riverside populations. Three hundred eighty-eight individuals, selected after inclusion and exclusion criteria were studied. The genotype of apolipoprotein E more frequent was ɛ3/ɛ3, followed by ɛ3/ɛ4. Allelic frequencies were 0.043: 0.784: 0.173 to ε2: ɛ3: ɛ4, respectively. The median level of total mercury in hair was 4.2 μg/g (1.9- 10.2). A significant proportion of participants (24.8%) had mercury levels above 10 μg/g, as recommended by the WHO limit, and 12.8% of participants showed a total content of mercury greater or equal to 20 μg/g. Interestingly, Tucuruí levels were much higher than levels in the Tapajós (area recognized by the presence of mining gold activity). We identified 29% of patients with ApoE4 (considered at risk) and 8 maximum risk individuals (carriers of ApoE4 and mercury content above the limit of 10 μg/g). Moreover, there was a significant difference in mRNA levels of S100B protein between groups exposed to high and low levels of mercury. For the first time, markers of the three spheres of influence in human toxicology (exposure, susceptibility and poisoning) were studied. Our data already support the use of these markers to monitoring the Amazonian populations. This knowledge will assist the development of prevention strategies and making government decisions facing the problem of the impact of the mercury in the Amazon.
Acesso aberto (Open Access)
Análise de patógenos orais entre indivíduos portadores de câncer gástrico e indivíduos sem câncer
(Universidade Federal do Pará, 2021-06) OLIVEIRA, Gyselle Ribeiro de Carvalho; BURBANO, Rommel Mario Rodriguéz; http://lattes.cnpq.br/4362051219348099; https://orcid.org/0000-0002-4872-234X
The loss of teeth and lack of oral hygiene have been associated with the risk of developing gastric cancer in several populations evidenced in epidemiological studies. In this study, we quantitatively compared the proportion of oral pathogens in individuals with gastric cancer and individuals without cancer in a referral hospital in the city of Belém, Brazil. This study evaluated 192 patients with gastric cancer and 192 patients without cancer. Periodontal clinical examination was performed, and all individuals were submitted to the collection of salivary and dental biofilms. When comparing the median periodontal indexesin the gastric and cancer-free groups, it was statistically significant in the gastric cancer group compared to the probing depth of the periodontal pocket. Levels of bacterial DNA were observed in saliva and dental plaque, with a statistically significant difference between individuals with cancer and without neoplasia in all the bacteria surveyed. Significant relationships between biological agents and gastric cancer have been found in bacterial species that cause high rates of periodontal pathology and caries. The results suggest a different quantitative association in the presence of oral pathogens between individuals without cancer and patients with gastric cancer. As noted, it cannot be said that the bacteria present in the oral cavity increase the risk of gastric cancer or are aggravating factors of the disease. However, it is worth mentioning that, as it is part of the digestive system, the lack of care for the oral cavity can negatively affect the treatment of patients with gastric cancer.
Acesso aberto (Open Access)
Análise de variações genômicas em genes da região cromossômica 22q11.2 em pacientes esquizofrênicos do Estado do Pará
(Universidade Federal do Pará, 2015-08-29) MORAES, Leopoldo Silva de; BURBANO, Rommel Mario Rodriguéz; http://lattes.cnpq.br/4362051219348099
The COMT Val158Met and ZDHHC8 rs175174 polymorphisms have received increased attention in the molecular study of schizophrenia not only because they are localised to the main susceptibility locus of the disease, 22q11, but also because they are related to the dopaminergic status of the prefrontal cortex and the activity of several neuronal proteins, respectively. To evaluate the influence of the polymorphic genotypes on schizophrenia, we used real-time PCR to genotype 130 patients and 175 controls in a population from the North Region of Brazil. Our results indicated an absence of association between both polymorphisms and the likelihood of schizophrenia in the population studied. However, when categorised by gender, we found a dichotomous association between the Met/Met genotype of the COMT Val158Met polymorphism and susceptibility to schizophrenia, conferring a higher probability of disease in men (OR = 10.76; CI 95% = 2.09–55.34; p = 0.004) than in women (OR = 0.23; CI 95% = 0.07–0.69; p =0.009). Moreover, the variance analysis showed an association of the genotypes Val/Met (COMT Val158Met) and GG (ZDHHC8 rs175174) with higher average age at onset of schizophrenia. Our study supports the hypothesis of a gender-dependent association of the COMT Val158Met polymorphism with schizophrenia, in addition to suggesting an influence of both polymorphisms studied on the age at disease onset.
Acesso aberto (Open Access)
Análise do efeito tóxico e alterações transcriptomicas de células neuronais e gliais após exposição ao fluoreto
(Universidade Federal do Pará, 2019-05-23) GOMES, Bruna Puty Silva; OLIVEIRA, Edivaldo Herculano Corrêa de; http://lattes.cnpq.br/0094007714707651; LIMA, Rafael Rodrigues; http://lattes.cnpq.br/3512648574555468
Despite being widely used in dentistry for dental carie control, in high amounts fluoride may be associated with side effects of which the best known is dental fluorosis. In addition, studies suggest that even at low concentrations fluoride may exerts toxicity leading to damage on CNS. Functional toxicogenomics analysis of gene profile after exposure to contaminants has been used as a tool for the identification of biomarkers of exposure, as well as for the identification of signaling pathways that may be used for treatment and / or prevention of damage caused by the toxicity of certain compounds. As the molecular mechanism of fluoride toxicity still unknow, analysis of F chronic exposure on gene expression profile of CNS cells are necessary. Here we aimed to show the effect of fluoride exposure of plasma concentration founded on population that used to be exposed to fluoridated drink water, on the main CNS cells. In this way, we have used human cell lineage IMR-32 (neurons) and U87 (glial cells) to analyze parameter of viability, morphology and cell metabolism, ATP-synthesis, oxidative stress, DNA damage and global gene expression profile after 10 days exposure. Our results have shown that fluoride does not induce changes in IMR-32 cells. On the other hand, it induces cell death by necrosis, increased metabolism, decrease in ATP and GSH / GSSG in U87 cells and DNA fragmentation. The U87 gene expression profile is differentially altered after fluoride exposure, decreasing 1735 genes and an increasing expression of 1047 genes after exposure to 0.095μg / mL and decreasing of 1863 gene expression and increasing of 1023 expression after exposure to 0.22μg / mL. We also highlighted the major molecular pathways altered after exposure, such as the signaling pathway TNF-alpha via NFK-B and mitochondrial process. We also showed genes with significant importance biology (hub genes) such as the genes PTGES3, EP300, CYP1B1, RPS27A. Our results suggests that glial cell are affected by fluorides exposure and mitochondria has a major role on the mechanism of fluoride toxicity.
Acesso aberto (Open Access)
Análise do perfil do número de cópias e transcriptoma de pacientes com gliomas e em linhagens de glioblastomas tratadas com pisosterol
(Universidade Federal do Pará, 2018-10-17) FERREIRA, Wallax Augusto Silva; OLIVEIRA, Edivaldo Herculano Corrêa de; http://lattes.cnpq.br/0094007714707651
Central Nervous System Tumors (CNS) account for approximately 2% of all cancers. Although the incidence of CNS tumors is small, compared to other neoplasms, these tumors are among the most serious human malignancies because they affect the organ responsible for the coordination and integration of all organic activities. Gliomas represent approximately 80% of all intracranial tumors, typically affecting adults, with a high incidence between 40 and 65 years of age. Although numerous anti-glioma drugs have already been developed, they induce adverse reactions and their therapeutic effects are not satisfactory. The objective of this study was to evaluate and compare the profile of Copy Number Variation (CNV) and gene expression of patients diagnosed with gliomas and in glioblastomas cell lines (U87-MG, U343, AHOL1 and 1321N1) treated with pisoterol. For rhe experiments done with the cell lines treated with pisoterol, we demonstrated that they were highly sensitive to pisoterol treatment. This drug reduced the number of live cells in a dose-dependent manner. In addition, we demonstrated that after 48h of exposure to pisoterol, all cell lines were blocked in G2/M. Finally, we demonstrate that the pisosterol can modulate the expression of several genes of ATM/ATR pathway, promoting apoptosis. We demonstrated on genomic scale that all the cell lines had more genes that were significantly down-regulated than up-regulated after the treatment with pisosterol. For the experiments done with the gliomas biopsies, we demonstrated that only 11 genes (TNFRSF1A, SNAPC2, CASP8, IRAK3, GPX3, FZD9, TFAP2C, CDH1, RPRM, POU4F3 and MGMT) exhibited changes in the pattern of methylation in all grades analyzed. In addition, the methylation pattern of these 11 genes had correlations with some clinicopathological characteristics, such as age, sex and histological grade. And finally, we made a molecular characterization describing the CNVs of the gliomas originating from Belém-PA.
Acesso aberto (Open Access)
Análise genômica comparativa e os polimorfismos nos genes TNFA, IFNG IL6 e IL10 associados à expressão de citocinas na infecção por Plasmodium vivax no município do Itaituba, Estado do Pará
(Universidade Federal do Pará, 2018-07-19) PIMENTA, Tamirys Simão; MACHADO, Ricardo Luiz Dantas; http://lattes.cnpq.br/0307356330748427; OLIVEIRA, Edivaldo Herculano Correa de; http://lattes.cnpq.br/0094007714707651
In endemic areas of Asia, Oceania, Central and South America and in the horn of Africa P. vivax malaria is a major cause of morbidity with 35 million cases annually. In Brazil, the Amazon region concentrates almost all cases and infections registered countrywide, with more than three hundred thousand cases per year. Several evidences suggest that an exacerbated inflammatory response associated to density parasite is likely to aggravate the malaria symptoms. We assessed the haematological and immunological aspects, genetic alterations related to CNVs that could lead to phenotypic alterations, conferring resistance or susceptibility to malaria, as well the presence of polymorphisms in cytokine genes and their association with the infection in patients living in a gold-mining area in a gold-mining in the Brazilian Amazon Region, establishing patterns of immune response characteristic of primary malaria, recurrent malaria and endemic control. Six SNPs (TNFA-308G/A, IFNG+874T/A, IL6-174G/C, IL10-1082G/A, -819C/T, -592C/A) in four genes were determined; blood cell count was conducted on automatic analyzer; plasmatic cytokines IL-6, IL-10, TNF-α and IFN-γ were quantified by flow cytometry and density parasite was estimated by thick blood films with confirmation by nested-PCR; the CNV was estimated by aCGH and association between copy number and phenotypes (parasite load, mean number of clinical infections of malaria and gender) was assessed. The statistical analyzes were performed by Graph-pad prism 6.0 and Bioestat 5.0. No significant association was found between SNPs and malaria infection; cytokine levels were higher in malaria group when compared to endemic control; production of IL-10 was higher in the presence of GCC/GCC haplotype; IFN-γ levels were correlated with previous malaria episodes; malaria patients showed lower platelet numbers, reduction on white blood cells count and an increased monocyte percentage; significant increase in the IL-6 and IL-10 plasmatic levels in both malaria groups; the primary malaria patients displayed the highest significant plasmatic IFN-γ levels; recurrent malaria patients displayed the highest significant plasmatic TNF-α; malaria infection demonstrated correlation between parasite density and TNF-α, IL-6 and IL-10 levels; a total of 112 amplified genes and 12 deleted genes were observed and the CNVs found did not include any gene related to receptors or vivax malaria resistance factors. There were no statistically significant correlations between the clinical and pathological data (parasite load, mean number of clinical infections of malaria and gender) and the presence of CNVs in the patients studied. This study provides additional data on Plasmodium-host immune response and describes the quantitative changes in the human genome in P. vivax infection in an endemic area of garimpo.
Acesso aberto (Open Access)
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).