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Tese 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/0676220027193876Cancer 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 (CT) and 3693 (GA) of mitochondrial DNA, had no associated pathological record and were related to population markers. The AG 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.Dissertação Acesso aberto (Open Access) Análise citogenética como bioindicador para pacientes com diagnóstico sugestivo de Alzheimer(Universidade Federal do Pará, 2013-08-02) NEGRÃO, Igor Patrick Ramos; BAHIA, Carlomagno Pacheco; http://lattes.cnpq.br/0910507988777644; PEREIRA JÚNIOR, Antônio; http://lattes.cnpq.br/1402289786010170Alzheimer's disease (AD) is a neurodegenerative disease that causes neuronal death and consequent progressive loss of cognitive functions, reducing the capacity for work, interfering with social relationships and behavior of the patient. Among the diseases that cause dementia, AD is the most frequent nature of the vascular a ratio of 4:1, respectively. In addition to the pharmacological therapies, diagnostic methods assist in the early identification of the disease by helping the pretreatment, thus reduced disease progression. Currently cytogenetic studies have demonstrated chromosomal abnormalities in individuals with AD and may aid in the diagnosis of disease. The aim of this study was to investigate the potential of karyotype analysis of peripheral blood lymphocytes as a diagnostic biomarker of Alzheimer's disease. For this work, we used two groups of women aged 65 or more, one group (10) suffering from AD and other normal group (10). Each subject was submitted to the socioeconomic survey, a cognitive screening test (MMSE) and the Venous blood lymphocyte culture and chromosome analysis. Our results demonstrate that the group of women with AD showed high rate of monosomy and trisomy compared to normal women. Through the study of history via questionnaire, we found the lifestyle of both groups. Compared the relationship of chromosomal abnormalities with the cognitive level of the AD group, we evidenced an inverse trend between the number of monosomy / trisomy and cognitive performance. Another aspect of our analysis was the role of each chromosome linked to AD. Chromosomes 1, 14 and 21 showed no trisomy and verify the frequency of monosomy, each chromosome has frequency below 3 % of aneuploidy, i.e., the chromosomes studied did not have a great importance in chromosomal alterations found in the study.Dissertação Acesso aberto (Open Access) Análise da interação in vitro entre Fonsecaea pedrosoi e macrofagos peritoneais de camundongos C57/BL6 e BALB/c(Universidade Federal do Pará, 2008-04-02) YAMANO, Suellen Sirleide Pereira; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125Chromoblastomycosis (CBM) is a chronic, subcutaneous, granulomatous infection caused by traumatic implantation in the skin of several dematiaceous fungi, usually Fonsecaea pedrosoi. Brazil has second highest disease prevalence in the world and Para State is the most endemic area. Histologically, CBM is characterized by the presence of multinucleated giant cells and sclerotic cells can be found engulfed by macrophages. The objective of this study was to analyze the different aspects of interaction between peritoneal macrophages from BALB/c or C57/BL6 mice with F. pedrosoi conidia or sclerotic cells, calculating infection, phagocytosis and cellular fusion rates. The results showed phagocytosis and infection rates with conidia higher than sclerotic cells to BALB/c (p <0.05), while the rate of cellular fusion was higher for sclerotic cells interaction, with Langhans giant cells formation, in comparison to foreign-body giant cells after interaction with conidia. Macrophages from BALB/c co-cultured with conidia produced more TNF-α than control group after 3 to 72 hours, and more IL-10 after 3h. Macrophages interacting with sclerotic cells produced more TNF-α than control group after 1h and 3h, and the amount of IL-10 was higher after 72h of interaction. In the co-culture of C57/BL6 macrophages with conidia, the presence of large vacuoles after 24h was observed, while in the coculture with sclerotic cells, macrophages were detached from coverslip glasses after 24 h. Our results indicate higher levels of TNF-α after interaction of conidia compared to controls at 1 and 72 h and increase of IL-10 after 48h. However, after interaction with sclerotic cells, only IL-10 differed from control, being higher after 1 to 48 hours. All of these data suggest that macrophage response to fungus is different between BALB/c and C57/BL6 mice, differing also on the response of the same type macrophage for each fungal form, sclerotic cells apparently being more immunogenic than conidia.Tese 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/0094007714707651In 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.Tese Acesso aberto (Open Access) Aspectos bioquímicos e morfológicos da matriz extracelular das cartilagens septal e alares do nariz suíno(Universidade Estadual de Campinas, 1999-07-02) MESSIAS JÚNIOR, Nazário de Souza; GOMES, Laurecir; http://lattes.cnpq.br/0032653238117133The objective of the present study was to characterize some aspects of the extracellular matrix components of the alar and septal cartilage of the nose of swine and to examine the histological and histochemical aspects of the two types of cartilage. The kaotropic agent Gu-HCl was used to extract the matrix components. The predominant glycosaminoglycan in the two caitilages was chondroitin sulfate. After ultracentrifugation, the D4 fraction of the septal and alar cartilages was fractioned on DEAE-Sephacel using na NaCl gradient. SDS-PAGE analysiys showed similarities between the two cartilages, with the presence of fíbromodulin, decorin and binding proteins. Only the septal cartilage presented the 115 kDa component, which possibly appears as 30 kDa subunits after the action of the reducing agent. The glycosaminoglycan populations were determined by fractionation on Sepharose CL-6B followed by analysis of the samples on the agarosepolyacrilamide gel. The two cartilages presented a strongly polydispersed population, which was less uniformly polydispersed in the septal than in the alar cartilage. AD2 fractionation on DEAE-Sephacel and sample analysis by SDS-PAGE and agarosepolyacrylamide electrophoresis showed the presence of the small proteoglycans biglycan and decorin, possible associated with the large proteoglycans. The septal cartilage presents chondrocytes roughly arranged in rows accompanyng the widest axis of the cartilage, whereas the alar cartilage presents chondrocytes characteristically arranged in isogenic groups. Histhochemistry was performed usisng Xylidine Ponceau for total proteins, Picrosirius-Hematoxylin for collagen, Toluidine Blue for total glycosaminoglycans. And alcian blues, pH 2.5 and pH 1.0, for sulfated and carboxylated glycosaminoglycans. Total glycosaminoglycans are more cleary visible in the territorial matrix, whereas total proteins are more cleary visible in the interterritorial matrix. The collagen of the septal cartilage is roughly organized along the longitudinal axis of the specimens between the chondrocytes rows.Dissertação Acesso aberto (Open Access) Avaliação antitumoral de amidas graxas derivadas de triglicerídeos de óleo de andiroba (Carapa guianensis aublet) em linhagem celular de glioma in vitro(Universidade Federal do Pará, 2022-06-26) SILVA, Nágila Monteiro da; OLIVEIRA, Fábio Rodrigues de; http://lattes.cnpq.br/4538804050936779; https://orcid.org/0000-0003-2761-3440; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978; https://orcid.org/0000-0003-3647-9124Glioma is a rare type of tumour, which acts on Nervous System in a very aggressive way, presents problems in its diagnosis, low effective treatments and survival time less than one year after diagnosis. Due to factors such as intratumoral cell variability, inefficient chemotherapy drugs, adaptive resistance development to the drugs and tumour recurrence after resection, the development of new drugs becomes necessary. In this sense, molecules analogues to endocannabinoids such as fatty amides are a good alternative, since scientific literature shows that they can act as antitumor agents through the interaction with the endocannabinoid system, which modulates many metabolic pathways related to cancer. In this work, two fatty amides synthetized from andiroba (Carapa guianensis aublet) using lipase from Candida antarctica-B (CAL-B) oil were tested aiming to evaluated its potential in the glioma treatment in vitro (C6). AGs reduced C6 cell viability in a dose dependent manner while were not toxic to normal glia cells. Both FAA1 and FAA2 caused apoptosis cell death and also loss of mitochondrial integrity probably by inhibiting PI3k/AKT pathway. Furthermore, FAAs were capable of reduce the C6 migratory potential. In conclusion FAAs have a promising potential to treat glioma-type brain cancer.Dissertação Acesso aberto (Open Access) Avaliação da atividade antineoplásica dos fármacos metformina e mebendazol isolados e em associação em linhagem celular de câncer(Universidade Federal do Pará, 2018) SILVA, Karla de Assis; KHAYAT, André Salim; http://lattes.cnpq.br/6305099258051586Gastric cancer will account for 782,685 deaths worldwide in 2018, being the fifth most common cause of cancer in the world and a fourth in Brazil. For example, the diagnosis of pathognomonic cases, the diagnosis of gastric cancer occurs late in most cases. In addition, this is a series of patients with chemotherapy and radiotherapy, being a resection surgery that offers the greatest healing potential. Adenocarcinoma is the most common subtype of lung cancer, with incidence greater than 90%. The risk factors for the pathology are multiple and cover the genetic, environmental and food. The drugs metformin and mebendazole, now used in the processes of diabetes and parasitic infections, are presented as antineoplastic effects in studies of various types of cancer. For metformin, it is possible that anticancer mechanisms of action, among others, may be made viable by LKB1 / AMPK / mTOR. Mebendazole prevents the polymerization of tubulins, inhibiting the growth and invasiveness of cancer cells. Thus, the present study has as a preventive effect metformin and mebendazole, known for their anti-neoplastic antibodies and low toxicity, in isolation and in combination with AGP01 (prescribed from neoplastic cells present in the patient with gastric ascites fluid of the intestinal type). This study is in vitro: MTT cytotoxicity, viability / apoptosis and necrosis assessment, cell cycle analysis and migration assay. One metfomine had 6.2mM IC50 and the 300mM IC50 mebendazole given alone, when combined with a new level of new IC50 values of 1.8mM and 88nM, respectively. The cell migration was inhibited for the metformin from the time of 12h and mebendazole from the time of 24h, a combination of the drugs showed no change in the time of inhibition, but increased the reliability of the test. Mebendazole and metformin induced cell death by apoptosis and prevented cell cycle progression, increasing the percentage of cells in the G1 / G0 phase and decreasing the percentage of cells in the S phase and G2 / M phase. These data confirm, at least in the antineoplastic effects of these drugs.Tese Acesso aberto (Open Access) Avaliação da integridade genômica mitocondrial em gliomas de alto e baixo grau na população paraense(Universidade Federal do Pará, 2016-12-14) COSTA JÚNIOR, Carlos Antonio da; BORGES, Bárbara do Nascimento; http://lattes.cnpq.br/0676220027193876Cancer is characterized by fast abnormal cells proliferation which grow beyond their normal limits and may invade adjacent or distant tissue. Cancer CNS represents 2% of all cancers in the world, being slightly higher in men than in women. Mitochondria are responsible for producing most of the cellular ATP by oxidative phosphorylation (OXPHOS), may also act through glycolysis for same purpose, not requiring only oxygen. This option is a particular cancer cell property, also known as the Warburg effect. One hypothesis to explain this metabolic change may be related to mitochondrial DNA (mtDNA) defects caused by OXPHOS where these mutations can lead cancer cells to glycolysis. Eight mtDNA regions (D-LOOP, ND1, ND3, CO I, CO II CO III, ATPase 6 and ATPase 8) were analyzed in patients’ neoplastic tissues with glial cell cancer in Pará population, relating the data with the pathological and clinical characteristics of the patients. Among the changes found, the complex I seem to be decisive for the progression of high-grade tumors, as well as changes indel seem compromising important structures for OXPHOS. Deletions 4977 bp, when combined with other changes in ND1 / ND3 or heteroplasmias suggest poor prognosis, however, seem to have a reduced risk when changes in ND1 and ND3 are simultaneous.Dissertação Acesso aberto (Open Access) Avaliação da seletividade olfatória causada pela infecção por SARS-COV-2(Universidade Federal do Pará, 2022-09-16) ALMEIDA, José Ramon Gama; BASTOS, Gilmara de Nazareth Tavares; http://lattes.cnpq.br/2487879058181806; ÁVILA, Paulo Eduardo Santos; http://lattes.cnpq.br/4673218055614655Introduction: Sudden loss of smell is one of the most prevalent symptoms of COVID-19. The sense of smell ranges from detecting warning odors in the environment to building our most pleasurable experiences. This sense stimulates a complex neural network, including the temporal lobe, the amygdala, the insula and a large part of the limbic lobe: the loss of smell should not, only, be considered a sensory symptom, but also a psycho-sensory syndrome. Objectives: This study was aimed to evaluate the olfactory selectivity and trigeminal sensation in olfactory alterations reported by patients diagnosed with COVID-19. Methods: The Randomized case-control study involving 88 individuals: COVID-19 without olfactory dysfunction previously diagnosed prior to the pandemic period; with COVID-19 with olfactory dysfunction diagnosed before the pandemic period; and without COVID-19 and loss of olfactory sensitivity during the pandemic period, with persistent anosmia or hyposmia after SARS-CoV-2 infection. All individuals participating in the study (Control/intervention groups) diagnosed with or without COVID-19 underwent evaluation data collection form and psychophysical sensory test which included olfactory test, olfactory memory test, olfactory threshold test and trigeminal sensation test (CEP-UFPA: 40962420.2.0000.0018). ANOVA follow the Tukey test. Results: The results demonstrated which all patients with or without COVID-19 diagnosis, loss the ability to not identify the odorant of banana essence when compared to the healthy health group being significant control vs. COVID-19 without p<0.0002 and significant control vs. COVID-19 with p<0.0010 and not significant COVID-19 without vs. COVID-19 with p>0.05. For the short-term olfactory memory test which all patients with or without COVID-19 diagnosis, demonstrated an increase in the misidentification of odorants presented when compared to healthy controls, as well as in the olfactory threshold differences in perception were observed between the groups. analyzed. The absence of at least one chemosensory function (cooling sensation) of the trigeminal during the test period was reported by which all patients with or without COVID-19 diagnosis, when compared to the healthy control group. Conclusion: In this way, SARS-CoV-2 infection may be promoting an olfactory dysfunction that affects the perception of banana odor, as well as the affected long-term characteristic such as olfactory memory, olfactory threshold and trigeminal sensation of olfactory loss that COVID-19. 19 generates in patients may provide clues to therapeutic interventions aimed at preventing, alleviating or curing long-term olfactory dysfunction in COVID-19.Tese Acesso aberto (Open Access) Avaliação do efeito da fração lipídica extraída de Agaricus brasiliensis antioxidante e imunomoduladora in vitro e em modelo de sepse letal em murino(Universidade Federal do Pará, 2021-01) LIMA, Kely Campos Navegantes; MONTEIRO, Marta Chagas; http://lattes.cnpq.br/6710783324317390; https://orcid.org/0000-0002-3328-5650Sepsis is defined as a potentially fatal organ dysfunction caused by a dysregulated host immune response to an infection. During sepsis, dysregulation of the host response occurs with the excessive release of pro-inflammatory mediators, generation of reactive species with depletion of antioxidant defenses and cellular damage. As a result, the patient develops organ dysfunction. In this context, our group proposes the A.brasiliensis Lipid Fraction (FLAb) as a possible therapy for sepsis considering its immunomodulatory and systemic antioxidant activity in a murine sepsis model. Thus, the present study aimed to evaluate the activity of FLAb isolated in vitro and to evaluate the effect of treatment with FLAb alone or associated with the antibiotic ertapenem (F-Erta) on coagulation, antioxidant and immunomodulatory parameters in the lethal sepsis model in murine. For this, FLAb was kindly provided by Dr. Herta Dalla- Santa from UNICENTRO. In the present study, the antioxidant capacity of different concentrations of FLAb (1.25 and 5 μg/mL) was evaluated and in a RAW 264.7-Luc macrophage cell line, cytotoxicity, phagocytic capacity, nitric oxide, NF-κB activity and cytokines TNF-α and IL-6 were evaluated.The survival rates were analyzed 7 days in a model of CLP sepsis in swiss albino mice (Mus musculus), and treated with CLP+Salt (0.9%), CLP+FLAb (0.2mg/Kg), CLP+F-erta (0.2mg) /Kg; 30mg/Kg). For evaluation of on coagulation, antioxidant and immunomodulatory parameters, the mice were treated by 6 and/or 24h after CLP. In vitro, FLAb show antioxidant and anti-inflammatory activity in both concentrations. In vivo, all CLP+Salt animals died within a maximum of 48 hours while the FLAb and F-Erta treated groups survived the 7 days. During this period, clinical parameters of these animals were evaluated, the septic animals treated with saline showed piloerection, with little active level of consciousness and most of the time they were stopped in the cage, some of them had ocular secretion. In addition, animals treated with saline showed significant weight loss, reduced water and feed consumption resulting in death. The FLAb and F-Erta groups were active, with normal appearance, with normal breathing and heart rate, in addition to consuming water and food within normal limits. In the inflammatory site, peritoneal cavity, the treatment with FLAb showed an anti-inflammatory effect, decreased reactive oxygen species (ROS) and increased GSH antioxidant activity and protected from cell damage, maintaining neutrophil recruitment and nitric oxide levels (NO), reducing the bacterial load. Regarding coagulation parameters (platelet count, tp and ttpa), treatment with FLAb and F-Erta eliminated the bacterial load and protected the animals from tissue damage. In the liver, 6 hours after CLP the treatment with FLAb and F-ERTA was observed in the biochemical parameters protective effect, in addition, it presented immunomodulatory, antimicrobial and antioxidant activity preventing liver damage. In the parameters evaluated in the heart, the treatment with FLAb and F-ERTA after CLP protected the animals from cardiac damage through immunomodulatory, antimicrobial and antioxidant activity. In this sense, FLAb alone showed promise as a treatment and/or adjunct in sepsis, in addition to preventing organic dysfunction in septic animals.Dissertação Acesso aberto (Open Access) Avaliação do método de fotooxidação do DiI por diodo emissor de luz (led): aspectos morfológicos de células horizontais da retina humana(Universidade Federal do Pará, 2013-11-25) FARIAS, Flávia Moura Gaia; SILVA FILHO, Manoel da; http://lattes.cnpq.br/2032152778116209The analysis of cell morphology is a crucial aspect of neurobiology, since the relationship between form and function can define the physiological processes in health and disease. One of the main methods to evaluate cell morphology is through photooxidation of intracellular fluorescent markers, among which is the perchlorate of 1,1 '-dioctadecyl-3, 3,3', 3'-tetramethyl-indocarbocianina (DiI) a lipophilic carbocyanine that can make living or fixed cells. The DiI was chosen for this work because, among other factors, it is used for important studies of cell morphology. As a model for evaluating the quality of photooxidation of the apparatus constructed for this purpose, horizontal cells of human retina were elected in order to proceed with morphometric study these cells further, in view of the lack of studies with this approach in human retina. This study evaluated the quality of the DiI method photooxidation via LED and used the horizontal cells of the human retina as a model. The material was obtained from the Eye Bank of Ophir Loyola Hospital and sequence dissected, labeled with DiI crystals and photooxided with the LED device. The images resulting from the new method of lighting fluorescent tracer showed photooxidation of high-quality detailed neuronal morphology, similar to the results obtained in reactions of convencional photoconversion with a microscope, which indicates that the device maintains the efficiency of the photooxidation for revealing fine details of cellular morphology, including the advantages of larger areas of tissue processing and a considerable cost reduction for the use of microscopy dispensed into the process.Dissertação Acesso aberto (Open Access) Avaliação in vitro da suscetibilidade antifúngica de conídios e células muriformes de Fonsecaeae spp(Universidade Federal do Pará, 2018-08-14) TAKAHASHI, Aline Barral; SILVA, Moisés Batista da; http://lattes.cnpq.br/5525661855611118; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125Chromoblastomycosis (CBM) is a fungal disease characterized by chronic cutaneous and subcutaneous lesions, due to the previous transcutaneous implantation of different black fungi, belonging to the family Herpotrichiellaceae, the main agent belonging to the genus Fonsecaea. The Amazon region is the main endemic area of Brazil and our state is the main endemic area in the country and second in the world. There is no standardization of treatment, drug therapy is used, with different regimens, mainly itraconazole. New azolics such as posaconazole and voriconazole began to be used, but with limitations due to the high cost, the use of physical methods such as cryosurgery, thermotherapy, laser therapy and both. The low cure rates and high rates of relapse of the disease may be related to the different treatment methods employed and the lack of a standardized regimen itself. The objective of this work was to evaluate the in vitro sensitivity of conidia and in vitro induced muriform cells of Fonsecaea spp. in the presence of different antifungal agents, to date there are no studies evaluating the action of drugs using the pathogenic form of the fungus, reinforcing the importance of this study, following the method established in the standard M38-A2, Clinical and Laboratory Standards Institute. The minimum inhibitory concentration for each drug was obtained after 5 days of interaction, posaconazole showed better in vitro inhibitory activity for conidia (CIMMG = 0.632μg / ml) and muriform cells (CIMMG = 1μg / ml). In the analysis of the fungicidal concentration, in both conidia and muriform cells, posaconazole presented lower value (CFMMG = 6.72μg / ml, in both). The fungus samples presented distinct times to differentiate their conidia in muriform cells, therefore we believe that although they belong to the same genus the fungi present physiological peculiarities still unknown. In the in vitro susceptibility tests PCZ had the lowest inhibitory and fungicidal concentrations in both conidia and muriform cells, thus investing in susceptibility testing was important to identify sensitive or resistant strains.Dissertação Acesso aberto (Open Access) Avaliação in vitro do potencial genotóxico e citotóxico do extrato do açaí (Euterpe oleracea) clarificado sobre a linhagem celular AGP01 (câncer gástrico)(Universidade Federal do Pará, 2024-01) SANTOS, Thiago Souza; BAHIA, Marcelo de Oliveira; http://lattes.cnpq.br/3219037174956649; BURBANO, Rommel Mario Rodríguez; http://lattes.cnpq.br/4362051219348099; https://orcid.org/0000-0002-4872-234XAçaí (Euterpe oleracea MART) is a fruit of great importance for the Amazon region in nutritional, cultural and socioeconomic terms. In recent years, açaí has been the subject of several studies due to its beneficial properties for health, including effects against tumor cells. Therefore, the present work aimed to evaluate in vitro the genotoxic and cytotoxic effects of the clarified extract of açaí juice in a human metastatic gastric cancer cell line (AGP01 cells). For comparison purposes, a non-transformed cell line of African green monkey renal epithelial cells (VERO cells) was used. The viability assay by resazurin reduction, the comet assay, the determination of cell death by differential fluorescent dyes and the wound healing migration assay were performed. A reduction in viability was observed only in the AGP01 line within 72h. There was no genotoxic damage or cell death (through apoptosis or necrosis) in any of the cell lines. However, açaí extract induced motility reduction in both cell lines. The reduction in cell viability and the induction of the anti-migratory effect in the AGP01 cell line opens perspectives for exploring the potential of Euterpe oleracea as an adjuvant in the treatment of gastric cancer.Tese Acesso aberto (Open Access) Avaliação in vitro dos efeitos citotóxicos e genotóxicos do antifúgico Fluconazol em linhagens de rim de macaco verde (VERO)(Universidade Federal do Pará, 2018-11-26) CORREA, Regianne Maciel dos Santos; BAHIA, Marcelo de Oliveira; http://lattes.cnpq.br/3219037174956649Fluconazole is a broad-spectrum triazole antifungal that is well-established as the first-line treatment for Candida albicans infections. Despite its extensive use, reports on its genotoxic/mutagenic effects are controversial; therefore, further studies are needed to better clarify such effects. African green monkey kidney (Vero) cell line were exposed in vitro to different concentrations of Fluconazole and were then evaluated for different parameters, such as cytotoxicity (MTT/cell death by fluorescent dyes), genotoxicity/mutagenicity (comet assay/micronucleus test), and induction of oxidative stress (DCFH-DA assay). Fluconazole was used at concentrations of 81.6, 163.2, 326.5, 653, 1306, and 2612.1μM for the MTT assay and 81.6, 326.5, and 1306μM for the remaining assays. MTT results showed that cell viability reduced upon exposure to Fluconazole concentration of 1306μM (85.93%), being statistically significant (P<0.05) at Fluconazole concentration of 2612.1μM (35.25%), as compared with the control (100%). Fluconazole also induced necrosis (P<0.05) in Vero cell line when cells were exposed to all concentrations (81.6, 326.5, and 1306μM) for both tested harvest times (24 and 48 h) as compared with the negative control. Regarding genotoxicity/mutagenicity, results showed Fluconazole to increase significantly (P<0.05) DNA damage index, as assessed by comet assay, at 1306μM versus the negative control (DI=1.17 vs DI=0.28, respectively). Micronucleus frequency also increased until reaching statistical significance (P<0.05) at 1306μM Fluconazole (with 42MN/1000 binucleated cells) as compared to the negative control (13MN/1000 binucleated cells). Finally, significant formation of reactive oxygen species (P<0.05) was observed at 1306μM Fluconazole vs the negative control (OD=40.9 vs OD=32.3, respectively). Our experiments showed that Fluconazole is cytotoxic and genotoxic in the assessed conditions. It is likely that such effects may be due to the oxidative properties of Fluconazole and/or the presence of FMO (flavin-containing monooxygenase) in Vero cells.Dissertação Acesso aberto (Open Access) Avaliação in vitro dos efeitos de miltefosina, orizalina e TC95 no crescimento de Entamoeba histolytica(Universidade Federal do Pará, 2011) ALVARENGA, Betânia Mara; BATISTA, Evander de Jesus Oliveira; http://lattes.cnpq.br/2206444845201080Amebiasis is characterized as a chronic, noninflammatory, afebrile which involves the colon wall and can invade the intestinal mucosa and spread to other organs, especially the liver. Entamoeba histolytica infection is treated with metronidazole, but this one presents side effects, so researches have been done to find more effective drugs with no adverse effects to the patient. In addition, testing new drugs may reveal important features of the parasite, such as the discovery of metabolic pathways or possible differences in their structure. In the present study, it was tested the amoebicidal effect of metronidazole, dinitroaniline oryzalin, a microtubule inhibitor, miltefosine, an alkylphosphocholine, that inhibits the membrane phospholipids syntesis, and TC95, a hybrid drug that presents the functional groups dinitroaniline and alkylphosphocholine. It was performed a culture with protozoan trophozoites, where the drugs and the control with diluents were added after 7 hours of culture. It was made a count of the viable trophozoites 12 hours after the addition of the drugs and the control at intervals of 12 to 60 hours. After analyzing the growth curves, it was found that the TC95 was the most effective drug of the three treatments, miltefosine had little inhibition and oryzalin was not effective. This is due to greater susceptibility of the parasite to membrane damage, with little effectiveness on microtubules inhibition.Dissertação Acesso aberto (Open Access) Avaliação in vitro dos efeitos genotóxicos e citotóxicos do fármaco dipirona sódica (Metamizol Sodium) em linhagem de rim de macaco verde africano (VERO)(Universidade Federal do Pará, 2016-10-27) GOMES, Lorena Monteiro; BAHIA, Marcelo de Oliveira; http://lattes.cnpq.br/3219037174956649The dipyrone or metamizole belongs to the family of the pyrazolones. It is one of the nonsteroidal anti-inflammatory compounds (NSAIs) most used, Brazil included, mainly due to its low financial cost. However, in some countries the sale of dipyrone is prohibited due to reported severe cases of agranulocytosis as a result of its use. Despite its high usage, studies showing genotoxic and cytotoxic effects of dipyrone in mammalian cells are scarce. Therefore, in the present study we will assess cell viability, genotoxic effects, cytotoxic effects (by apoptosis and necrosis induction) and the induction of reactive oxygen species (ROS) in VERO cells (a cell line obtained from red kidney of green monkey) exposed to dipyrone. Our results showed a significant reduction in viability of cells exposed to dipyrone by the MTT assay. A significant increase in damage index evaluated by comet assay was also observed, which indicate its genotoxic effects. In which concerns the cytotoxic effects of dipyrone, we observed a significant increase in the number of apoptotic cells using fluorescent dyes after 24h and 48 h of treatment with the drug. Ours results also showed that there was no significant difference in the induction of ROS generation after treatment of the cells with the drug assessed by the DCFH-DA technique. Thus, our work showed that dipyrone is both a genotoxic and cytotoxic drug to VERO cells in the assessed conditions.Dissertação Acesso aberto (Open Access) Avaliação neuropatológica da lesão estriatal em ratos machos e fêmeas da linhagem lister hooded induzida experimentalmente por microinjeções de endotelina-1(Universidade Federal do Pará, 2012-05-30) SANTOS, Ijair Rogerio Costa dos; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072The inflammatory response may exacerbate the process harmful after acute neural disorders. Sexual dimorphism generated by different hormonal attendance between male and female have demonstrated neuroprotective abilities endogenous opposite, showing a better preservation of the integrity of the nervous tissue in female, putatively due to the presence of gynoid hormones. However, there is no research work comparing how this difference might affect the inflammatory response during stroke. In the present study, we investigated the differences in acute inflammatory processes of sexual dimorphism in adult rats of both sexes subjected to acute ischemic injury induced by endothelin (ET1) in the striatum. Six experimental groups were designed: male animals survival 24 hours (n = 8); male survival of 72 hours (n = 8); male survival of 7 days (n = 8) and female 24 hours survival (n = 8), females 72 hours of survival (n = 8); female seven days of survival. Histopathologic analysis was performed in the general sections stained with violet cresila. Macrophages, astrocytes, and neurons were identified by immunohistochemistry with antibodies specific for these inflammatory cells (ED1, anti-GFAP and Anti-NeuN, respectively). We conducted counts activated microglia / macrophages and neuron bodies mentioned in the experimental groups. No difference was observed quantitatively between different sexes, but there was an apparent decrease in the amount of macrophages / microglia at 3 days but in males and in females, presenting apparently some difference in the activation of astrocytes was stronger in males. The results suggest that sex differences, at least in lineage Lister hooded is not enough to cause significant differences in the preservation of nerve tissue and in some aspects of the inflammatory response after induction of cerebral ischemia by ET1.Tese Acesso aberto (Open Access) Caracterização de aspectos morfológicos e ultra-estruturais do ciclo de vida de microsporidios encontrados em peixes da Região Amazônica(Universidade Federal do Pará, 2007-03-28) MATOS, Edilson Rodrigues; NASCIMENTO, José Luiz Martins do; AZEVEDO, Carlos José Correia de; http://lattes.cnpq.br/7216249286784978; http://lattes.cnpq.br/3066639837918744This work describes the results in the light (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM) obtained of the life cycle of some microsporidian species (phylum Microsporidia Balbiani, 1882) parasites of the ichthyofauna of the Amazon region. Emphasis special to the ultrastructural aspects of the different life cycle phases with evidence of the spores was given. The spores and life cycle stages characterized the different genus and species. The ultrastructural organization of the host tissues with the lysed aspects, that frequently occurred, and the ultrastructural aspects of the xenoma, was discussed.Dissertação Acesso aberto (Open Access) Citoproteção do ácido kójico (AK) na morte induzida por LPS em células de Muller de retina de embrião de galinha(Universidade Federal do Pará, 2017-12-07) CARVALHO, Giselle Cristina Brasil; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/72162492867849785-Hydroxy-2-hydroxymethyl-γ-pyrone (AK), a known inhibitor of tyrosinase, an enzyme important for melanin synthesis and therefore used for pigmentation disorders. AK also promotes significant activation of macrophages and promotes cytoplasmic accumulation of reactive oxygen species (ROS), suggesting its role as a potentiator of the immune system and microbicide. There is no work in literature that shows the action of AK in the central nervous system (CNS) as a cellular activator and its possible protective role against infections. To test this hypothesis, it use retinal Muller glia which have similar properties to those of macrophages. Therefore, the present work evaluates the action of AK as a possible protective role in LPS-induced cell death in culture of glial cells from chicken embryos. Cultures enriched with glial cells were treated with AK (10, 25, 50 and 100 μM) and LPS (0.1, 10, 100, and 500 ng / ml) for 24 hours. After treatment, the cells did not show AK-treated cytotoxicity; however, treated with LPS, cell death occurred in a dose-dependent manner. We verified the accumulation of EROs in groups treated with AK (100 μM) and LPS (100 and 500 ng / ml). Cultures co-treated with AK and LPS in the same concentrations there was a reduction of accumulation of EROs. AK was also able to inhibit the activity of antioxidant enzymes, (catalase and Superoxide dismutase) and glutathione levels, while LPS produces an increase in the activity of these antixodants. AK was able to inhibit the antioxidant enzymes and glutathione from the increase induced by LPS. These data show that AK promotes the modulation of oxidative and antioxidative balance as a possible protective mechanism in the cell death produced by LPS in Müller's Glia enriched cells.Tese Acesso aberto (Open Access) Diversidade de helmintos de Ageneiosus ucayalensis Castelnau 1855 (Pisces siluriformes) da foz do Rio Guamá e Baia do Guajará, Belém, Pará(Universidade Federal do Pará, 2010-05-10) GIESE, Elane Guerreiro; SANTOS, Jeannie Nascimento dos; http://lattes.cnpq.br/4543897195525368; VALLINOTO, Antonio Carlos Rosário; http://lattes.cnpq.br/3099765198910740Descriptive taxonomy studies of fish parasites in the Amazon Region are relevant, considering the great biodiversity of known hosts. The aim of this was to analyze parasites found in Ageneiosus ucayalensis, describing new helminthes species, redescribing species and finding new hosts for species of known helminthes, using Light Microscopy, Scanning Electron Microscopy and Molecular Biology. A. ucayalensis, a fresh water siluriform from South America, is poorly studied, despite its importance in the trophic chain of fresh water environment and the great value in the regular diet of Amazonian populations. Those hosts inhabit the base level of Guamá river and Guajará Bay being parasited by helminthes of Phyla Plathyhemintes, Acanthocephala and Nematoda, which are described for the first time. In this study two new species of Nematoda were described, Procamallanus (Spirocamallanus) belenensis and Cucullanus ageneiosus; new host and new occurrence were described for Procamallanus (Spirocamallanus) rarus; moreover, we described larval forms of nematoda of Families Anisakidae (Anisakis sp.), Cystidicolidae (Pseudoproleptus sp.) and Cucullanidae (Cucullanus sp.) parasiting A. ucayalensis. Parasites of phylum Plathyhelmintes, Class Cestoda, represented by metacestodes and adults of family Proteocephalidae were also described, as well as class Monogenea, represented by helminthes of family Dactylogiridae, subfamily Ancyrocephalinae and phylum Acanthocephala by specimens of Neoechinorhynchidae, family (Genus Neoechinorhynchus). We believe that the present study contributes with important data to parasite biodiversity from Amazon Region.
