Mestrado em Biologia da Relação Parasito-Hospedeiro (IPTSP)
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Item Mapeamento imunoinformático de regiões conservadas da proteína hexon de adenovírus humano(Universidade Federal de Goiás, 2020-03-17) Anjos, Déborah Carolina Carvalho dos; Souza, Menira Borges de Lima Dias e; http://lattes.cnpq.br/0054562567103606; Souza, Menira Borges de Lima Dias e; Fonseca, Simone Gonçalves da; Gardinassi, Luiz Gustavo AraújoHuman Adenoviruses (HAdVs) are important infectious agents associated with high incidence, morbidity and mortality in immunocompromised patients, such as those undergoing allogeneic hematopoietic progenitor cell transplantation (allo-TCPH). HAdVs belong to the family Adenoviridae, genus Mastadenovirus, and further classified into seven species (A-G) and 103 genotypes, characterized so far. The HAdV capsid consists mainly of the hexon protein, which has four highly conserved regions (CR1 - 4) that are known for their immunogenic potential, being one of the main targets of the T and B cell-mediated anti-HAdV immune response. The aim of the present study was to perform the mapping of potentially immunogenic epitopes, located in the HAdV hexon CRs and to evaluate the HAdV infection in patients undergoing allo-TCPH. To this end, predictions of T and B cell epitopes and IFN-γ induction were performed, considering 101 HAdV genotypes with sequences available in databases. The most conserved and best classified epitopes were then selected by the prediction programs to perform the molecular docking analysis with HLA alleles of the study population, consisting of nine adult patients undergoing allo-TCPH. It was also carried out the HAdV research by TaqMan real-time polymerase chain reaction (qPCR) in the patient's serum samples. As a result, regions containing overlapping T and B cell epitopes were obtained and, based on immunoinformatics analysis (prediction and molecular docking), two peptides with high conservation and immunogenic potential were designed. Nine patients were positive for HAdV by qPCR TaqMan, with the average viral load found in the serum samples of the study population being 6.71x1011 CG / mL. The genomic sequencing of the positive samples returned sequences that showed 100% similarity with sequences of the HAdV C hexon protein, deposited in a database. The present study allowed the mapping of the main immunogenic regions located in the CRs of the HAdV hexon, as well as the construction of two peptides that will be used in future studies to evaluate the immune response of patients undergoing allo-TCPH, participating in the present study.Item Escherichia coli, adenovírus e enterovírus em amostras de água consumida em áreas rurais de Goiás(Universidade Federal de Goiás, 2020-06-23) Lima, Fernando Santos; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Scalize, Paulo Sérgio; Santos, Mônica de Oliveira; Gabriel, Ellen Flávia Moreira; Gama, Aline RodriguesThe rural environment lacks basic sanitation services. Thus, the water supply and sewage disposal facilities are often under the initiative of each resident, who may not have the financial resources and technical knowledge to build and keep them functioning properly. Thus, water for human consumption is subject to fecal contamination and, consequently, to the presence of waterborne pathogens, such as enteric viruses. The objective of this work was to evaluate the fecal contamination of water samples from individual sources used for household supply in rural areas of the State of Goiás. The samples were collected from 86 homes, distributed in 15 communities, whose water sources were tubular wells, dug wells, springs, surface waters and rainwater. Escherichia coli (EC) bacteria, analyzed by the defined chromogenic substrate method, and the enteric virus human adenovirus (HAdV) and enterovirus (EV), analyzed by qPCR, were used as fecal contamination indicators. It was observed that 90.7% of the samples showed indications of fecal contamination. Detection rates were 74.4% for EC, 57% for HAdV and 9.3% for EV. The concentration averages of these indicators were, respectively, 8.34 x 101 NMP / 100mL, 8.6 x 105 CG / L and 9.75 x 105 CG / L. The EC indicator was the most prevalent in ground and surface water samples. The HAdV indicator was significantly more detected in groundwater samples than in surface water and was more efficient in indicating contamination in tubular wells. In cistern samples, viral indicators were the most prevalent. There was no association of frequencies or correlation of concentrations between EC and HAdV. HAdV indicated human fecal contamination and performed well as a complementary indicator. These results reveal that the analyzed population is vulnerable to waterborne diseases caused by enteric pathogens.Item Fluconazol e própolis co-encapsulados em nanopartículas mucoadesivas para o tratamento da candidíase vulvovaginal(Universidade Federal de Goiás, 2020-04-03) Silva, Jacqueline Teixeira da; Alves, Suzana Ferreira; http://lattes.cnpq.br/9799183444893510; Amaral, André Corrêia; http://lattes.cnpq.br/8801299423520104; Amaral, André Corrêia; Costa, Carolina Rodrigues da; Pereira, MaristelaVulvovaginal Candidiasis is the second most common infection in women of childbearing age, in about 70 to 90% of cases; the associated etiologic agent is C. albicans. The increase in the incidence of non-albicans species has led to the emergence of cases of resistance to antifungals, which is why the importance of new approaches in treatment. The objective of the present work was to develop and test in vivo a nanostructured system containing fluconazole and Green Propolis for the treatment of Vulvovaginal Candidiasis. The nanoparticles containing Fluconazole and Green Propolis were prepared following the technique of ionic gelation and presented satisfactory physical characteristics for the intended purpose. The association efficiency for the complexed compounds was performed using the technique of High Performance Liquid Chromatography (HPLC) and UV spectrophotometry. The Minimum Inhibitory Concentrations (MIC) for Fluconazole, Green Propolis and nanoparticles were determined, showing efficacy for use and fungistatic action on C. albicans ATCC 10231. Release analyzes of up to 72 hours with stabilization up to 96 hours were verified in the nanoparticles. The toxicity tests by hemolysis and Galleria mellonella, demonstrated positive results for propolis and negative for nanoparticles, suggesting safety in their use in a method with a murine model. In vivo tests were performed with an indication of efficacy for the proposed formulation.Item Co-encapsulação do trans-cinamaldeído e do fluconazol em nanopartículas de quitosana para o tratamento tópico da candidíase vulvovaginal(Universidade Federal de Goiás, 2020-04-03) Sousa, Paulo Henrique Dantas de; Amaral, André Corrêa; http://lattes.cnpq.br/8801299423520104; Amaral, André Corrêa; Alves, Suzana Ferreira; Silva, Luís Antônio DantasVulvovaginal candidiasis is an infection caused by fungi of the genus Candida. The increased incidence of vulvovaginal candidiasis maybe related to the advancement of chronic and immunosuppressive diseases along with the emergence of new mechanisms of fungal resistance. The objective of the present work was to prepare and characterize chitosan nanoparticles containing fluconazole and trans-cinnamaldehyde co-encapsulated for the topical treatment of vulvovaginal candidiasis. The empty nanoparticles, fluconazole, trans-cinnamaldehyde and coencapsulated were prepared using the ionotropic gelling technique. The nanoparticles were characterized as to their average size and polydispersity index by the dynamic light scattering technique. The surface charge of the nanoparticles was obtained by the electrophoretic migration technique. The empty nanoparticles had an average size of 448,9 ± 32,9 nm, PdI of 0,4 ± 0,1 and a surface load of 31 ± 0,4 mV, the nanoparticles of fluconazole exhibited an average size of 175,3 ± 8,9 nm, PdI of 0,2 ± 0,01, surface load of 33,2 ± 0,3 mV and encapsulation efficiency of 37,5% ± 3,1. Trans-cinnamaldehyde nanoparticles showed an average size of 352,6 ± 71,6 nm, PdI of 0,4 ± 0,05, surface load of 44,6 ± 3,7 mV and encapsulation efficiency of 44,6 % ± 3,7 and the coencapsulated nanoparticles had an average size of 234,5 ± 60,2 nm, PdI of 0,4 ± 0,07, surface load of 33,2 ± 0,5 mV and encapsulation efficiency of 56,8% ± 3 for fluconazole and 46,1% ± 5,1 for trans-cinnamaldehyde. The scanning electron microscopy of the nanoparticles presented an oval shape. The nanoparticles had their antifungal efficacy against the strain of C. albicans ATCC 10231 investigated by means of the minimum inhibitory concentration by the broth microdilution technique. The fluconazole nanoparticle showed antifungal efficacy with a minimum inhibitory concentration of 2 μg/mL and the trans-cinnamaldehyde was effective in the concentration of 75 μg/mL. The coencapsulated nanoparticle exhibited a minimum inhibitory concentration of 2 μg/mL for fluconazole and 37,5 μg/mL for trans-cinnamaldehyde. The coencapsulated nanoparticle exhibited the same minimal inhibitory concentrations as the fluconazole and trans-cinnamaldehyde nanoparticles. As for in vitro toxicity, nanoparticles were considered non-toxic. Regarding the in vivo toxicity test, only the empty nanoparticles exhibited a cytotoxic effect. All groups of nanoparticles proved to be effective in inhibiting fungal growth.Item Participação e regulação de macrófagos na malária experimental(Universidade Federal de Goiás, 2016-07-01) Tomé, Fernanda Dias; Nagib, Patrícia Resende Alo; http://lattes.cnpq.br/7172668220681444; Fonseca, Simone Gonçalves da; Celes, Mara Rubia Nunes; Nagib, Patrícia Resende AloMalaria is a neglected tropical disease caused by the protozoan parasite Plasmodium sp., which is capable of infecting red blood and liver cells. Profile Th1 and Th2 cells play an important role and can determine whether there is an effective response or increased sensitivity to the disease. Several factors may act on these cells and influence the immune response, such as leptin hormone, which in high concentrations stimulates Th1 cells and consequently M1 macrophages; on the other hand, leptin low levels may favor Th2 profile and M2 macrophages polarization. The macrophages role in the experimental fatal malaria is not clearly described. Thus, this work aimed evaluate the regulation of M1 and M2 macrophages during infection with P. berghei NK65 in vitro and regulatory mechanisms, leptin and cytokines in BALB / c mice. The animals were infected with iRBCs / P. berghei and euthanized at 4; 7 or 14 d.p.i. Every two days, the mice were weighed and parasitemia was estimated; it assessed the weight of the perigonadal fat, liver and spleen for index analysis thereof, as well as macroscopic of these organs. It was performed spleen and liver histological analysis and tissue extract was transferred to cytokine assay by ELISA. Serum leptin was determined by ELISA. M1 and M2 macrophages were differentiated and placed, in vitro, in the presence of infected mice erythrocytes to evaluate phagocytosis, elimination of the parasite, urea and NO production. Our data showed that the infection led to hepatosplenomegaly, fat loss and tissue damage. Initially infection by P. berghei led to increasing leptin and Th1 cytokines, followed by a immunoregulation and increasing IL-4 and IL-10 that characterize the regulation and Th2 polarization at 14 day after infection. In vitro, infected erythrocytes were able to induce a M2 macrophages regulation, causing the same to produce NO, a characteristic molecule of M1 macrophages. Despite the NO production, there was no decrease in infection rate, indicating escapement mechanism. These data together with previous data from our group show that the experimental malaria is able to induce immunomodulation and change macrophages profiles, and that this is due to polarization of cytokine response that is directly influenced by the leptin hormone; causing tissue damage and leading to high levels of parasitaemia, perpetuating the infection and subsequent death.