Doutorado em Medicina Tropical e Saúde Pública (IPTSP)
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Item Análise molecular e de qualidade de vida dos pacientes e familiares com xeroderma pigmentosum, residentes em Goiás, Brasil(Universidade Federal de Goiás, 2016-03-07) Souto, Rafael; Siqueira Júnior, João Bosco; http://lattes.cnpq.br/3644529827602550; Menck, Carlos Frederico Martins; https://orcid.org/0000-0003-1941-0694; Menck, Carlos Frederico Martins; Lacerda, Elisângela de Paula Silveira; Vêncio, Eneida Franco; Teles, Sheila Araújo; Brasil, Virginia ViscondeXeroderma pigmentosum (XP) variant is an autosomal recessive disease that involves changes in POLH. The study aimed to characterize the distribution of alleles mutated by Real Time PCR (RT-qPCR) in patients and families with clinical suspicion of XP, residents in Araras/Faina, State of Goiás. Additionally, we also, planned to evaluate the quality of life (QoL) by WHOQOL-Bref. In this community, the skin cancer incidence, due to this syndrome, is caused by mutation in the POLH gene, which encodes for DNA, polymerase eta, and two distinct mutations were detected, at the intron 6 e exon 8. Morover, at Trindade a different mutation was found in the same gene (intron 10). Molecular analysis by Real Time PCR (RT-qPCR) o 125 individuals at-tempted to identify the mutated alleles in POLH, which can result in disease and impact on quality of life. Of these, 29 clinically diagnosed as affected by XP syndrome, and 18 in the community of Araras/Faina and 11 are from other Goiás State locations. In Araras/Faina, of the 114 individuals analyzed, 12 were homozygous for the mutanted allele at the beginning of intron 6 (XPV 6/6), one homozygous for the mutanted allele at exon 8 (XPV8/8) and 5 are compound heterozygous for compounds two alleles (XPV 6/8). In addition, 36 patients were identified as carrying (as heterozygous) the mutation at intron 6 (XPV 6/wild-tipe) 12 carriers for muta-tion at exon 8 (8 XPV/wild-type) and 48 participants were wild type for the two alleles (XPV wild type/wild). In the study of 11 clinically affected patients and residents in other regions of the state of Goiás, 2 were positive for XPV with mutations in intron 10 (XPV 10/10) and 9 were negative for the three alleles identified in XPV. The Quality of Life evaluation gave relatively high scores when compared to the work of other groups that studied the Tourette syndrome, Wilson's disease and Thalassemia Major. In comparison using the Student t test between QoL scores of patients by XPV and not sick, it was obtained a p ≤ 0.05 for all domains of the WHOQOL-Bref, demonstrating that the XPV impacts the quality of life of those affected. However, even in a more stratified analysis , the comparison between QoL scores and genotypes for XPV, obtained a p ≤ 0.05 for the Physical and Environmental domains. Thus, we believe that molecular tests come uncovering cases of XP that were underreported showing the actual frequency of the syndrome in the state of Goiás, in addition, the measure of the perceived quality of life is showing the impact that these mutations promote affected in the XPV.Item Identificação por imunoproteômica dos exoantígenos do complexo paracoccidioides, com potencial aplicação no diagnóstico e terapia da paracoccidioidomicose(Universidade Federal de Goiás, 2019-08-26) Moreira, André Luís Elias; Bailão, Alexandre Melo; http://lattes.cnpq.br/5415221996976886; Bailão, Alexandre Melo; Borges, Clayton Luiz; Silva Neto, Benedito Rodrigues da; Rocha, Thiago Lopes; Oliveira, Milton Adriano Pelli deIdentification by immunoproteomic of exoantigens of the Paracoccidioides complex, with potential application in diagnosis and therapy of Paracoccidioidomycosis Fungi of Paracoccidioides complex are the etiological agents of Paracoccidioidomycosis (PCM), a systemic mycosis restricted to Latin American countries. Currently, the Paracoccidioides genus is represented by P. lutzii, P. americana, P. brasiliensis, P. restrepiensis and P. venezuelensis. In some cases, oral and skin mucosal lesions caused by other pathogens may coincide with lesions caused by Paracoccidioides spp.. Moreover, even with the advances in immunological techniques used for the diagnosis of fungal diseases, false-positive results rates for PCM are present. Thus, we investigated which antigens are secreted by 4 species of the Paracoccidioides complex in order to identify and characterize new molecules, thus increasing the spectrum of antigens to be used for future diagnostic tests of PCM. Through of nanoUPLC-MSE, 79 exoantigens were identified in 4 Paracoccidioides species. In silico analysis revealed 2 exoantigens exclusive to P. lutzii species, as well as the identification of 44 unique B-cell epitopes of the Paracoccidioides complex. Thirteen exclusive epitopes distributed among Paracoccidioides species also predicted, being this excellent molecules to be employed in the future for epidemiological studies. These results demonstrate a range of epitopes exclusive to the Paracoccidioides complex as well as the identification of molecules unique to each fungal species. In addition, these analyzes allowed the identification of new candidate biomarkers to PCM diagnosis, as well as the identification of molecules to be used as future epidemiological biomarkers.Item Carga epidemiológica da coqueluche e avaliação de impacto de vacinação de gestantes contra coqueluche implementada no Brasil em 2014(Universidade Federal de Goiás, 2022-08-10) Bagattini, Ângela Maria; Toscano, Cristiana Maria; http://lattes.cnpq.br/3772312631884265; Toscano, Cristiana Maria; Morais Neto, Otaliba Libânio de; Siqueira Júnior, João Bosco; Silva, Lara Lívia Santos da; Kuchenbecker, Ricardo de SouzaIntroduction: Pertussis is an acute infectious disease of respiratory transmission, with cyclical occurrence, it is endemic worldwide, represents an important global burden, particularly in children under one year of age who have more severe conditions and may progress to death. Between 2010 and 2014, a sudden and atypical increase in the number of cases of the disease was observed in several countries, including Brazil. There are several hypotheses about the factors associated with this resurgence, including disease cyclicality, replacement of whole-cell vaccines with acellular vaccines, falling vaccine coverage in children, and the effectiveness and duration of protection of vaccines in children, among others. Objectives: This study aims to characterize and estimate the epidemiological situation of pertussis in Brazil, evaluate the impact of vaccination of pregnant women with pertussis vaccine (dTpa) implemented in Brazil in 2014. Also, review the evidence about protection and duration of protection conferred by the whole cell vaccine against pertussis used for vaccination of children in the National Programs of Immunization. Methods: A study in three stages was carried out. The first stage described the epidemiological situation of whooping cough in morbidity and mortality in Brazil from 2000 to 2016. Next, the second stage with an interrupted time series ecological study was carried out with data adjusted by month and using the ARIMA model to assess the impact on cases and hospitalizations of children under five years of age with the introduction of the dTpa vaccine for pregnant women in 2014. The two stages used three independent health information systems, the Information System for Notifiable Diseases (SINAN), the Hospital Information System (SIH), the Mortality Information System (SIM) and the National Immunization Program Information System (SI-PNI), data were aggregated and evaluated by age group and federation unit. The third stage of the study was carried out through a systematic review to assess the effectiveness and duration of protection provided by the whole cell pertussis vaccine (DTPw) in children considering the vaccines currently available on the international market. Results: Pertussis showed cyclical patterns of disease burden over time between 2000 and 2016 in Brazil, in different regions with heterogeneous conditions, with a well-defined outbreak that started in 2011 and peaked in 2014, reaching mainly and more severely. children under six months, with 20,103 (54%), 19,919 hospitalizations (79%) and 565 deaths (93%). The incorporation of dTpa vaccination in pregnant women was associated with a significant reduction in pertussis cases and hospitalizations in children under six months of age when coverage is above 45%. After the introduction of dTpa, it is estimated that 2,124 cases and 1,439 hospitalizations for pertussis were avoided in children under six months of age in states with coverage above 45% between 2015 and 2016. Additionally, 12 studies with DTwP conducted between 2007 and 2020 were included for review, which have varied methodological quality and lack evidence on immunogenicity and duration of immunity indicating a short duration, less than five years. Conclusions: The analysis of different health information systems used showed consistent results throughout the period analyzed, reflecting the cyclicity of the disease and its resurgence from 2011. The incorporation of dTpa vaccination in pregnant women resulted in an impact on the reduction of hospitalizations and deaths of children when vaccination coverage above 45% is achieved. The wP vaccines currently in use have scant evidence of duration of immunity, even though they are used in over 100 countries, in most low- and middle-income countries. Relevance and Impact: The results of this work reinforce the importance of achieving and maintaining dTpa vaccination coverage in pregnant women above 45% in order to obtain a significant impact of vaccination of pregnant women in reducing hospitalizations for pertussis in children. Although wP is one of the most used vaccines globally in children in immunization programs in low- and middle-income countries, there has been an important change in the producers of these vaccines in recent decades, with large pharmaceutical companies having left the market and being replaced by producers in countries emerging technologies that today account for the totality of wP vaccines produced and used in the world. Evidence suggests that the duration of protection afforded by these vaccines in children is short. However, better quality evidence on the effectiveness and duration of immunity conferred by these vaccines is needed to support the definition of more appropriate vaccination strategies.Item Tendência da mortalidade por pneumonia em idosos no Brasil e o contexto da vacinação pneumocócica(Universidade Federal de Goiás, 2019-06-12) Miranda, Denismar Borges de; Andrade, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899; Morais Neto, Otaliba Libânio de; http://lattes.cnpq.br/4030124246791320; Moraes Neto, Otaliba Libânio de; Turchi, Marília Dalva; Souza, Maria de Fátima Marinho de; Bierrenbach, Ana Luiza; Sartori, Ana LúciaCommunity-acquired pneumonia is one of the most common infectious diseases and is considered a major cause of morbidity and mortality in the elderly population worldwide. Studies show the direct and indirect impact of 10-valent pneumococcal vaccine on hospitalizations for pneumonia in several countries. There is scarce knowledge regarding this impact on mortality in the elderly in the world and, to date, no evidence in the Brazilian population. This study aimed to propose models for correcting mortality rates due to pneumonia in the elderly in Brazil and to evaluate the indirect impact of PCV-10, introduced in the childhood immunization schedule, at these rates. This is a time-series study of mortality rates from pneumonia in the elderly from 2005 to 2016. For the time-series analysis, the models for predicting pneumonia rates, if the vaccine had not been implemented, were Seasonal Autoregressive Integrated Moving Average (SARIMA) in the presence of seasonality, Auto Regressive Integrated Moving Averages (ARIMA) when there was only trend, and Autoregressive Moving Average (ARMA) in the absence of trend and seasonality. Percentage difference between observed and predicted rates were calculated, considering statistical significance of 5%. There was an increasing trend of mortality due to pneumonia in the elderly in Brazil. Interrupted time series analysis showed that the estimated pneumonia mortality rates from the study were significantly lower than those predicted by the analysis models. There was probably an indirect impact of PCV-10 on the elderly. In the Southeast region there was a statistically significant difference between the rates observed and the predicted in the three age groups of the elderly.Item Epidemiologia molecular de isolados de toxoplasma Gondii na região metropolitana de Goiânia, Goiás, Brasil(Universidade Federal de Goiás, 2018-01-12) Rezende, Hanstter Hallison Alves; Vinaud, Marina Clare; http://lattes.cnpq.br/1921551651088660; Castro, Ana Maria de; http://lattes.cnpq.br/9232309971000621; Castro, Ana Maria de; Fernandes, Everton Kort Kamp; Rocha, Thiago Lopes; Cardoso, Cléver Gomes; Soave, Danilo FigueiredoThe protozoan Toxoplasma gondii is the etiological agent of toxoplasmosis and its definitive hosts are domestic cats (Felis catus) and other wild felids. Within the cities, stray cats are responsible for the dissemination of the parasite in the environment as they release oocysts which are the infective forms for humans and other animals. Backyard chickens (Gallus gallus) are in constant contact with the environment and feed directly from the soil. Therefore they are important indicators of environmental conditions. The knowledge regarding prevalence, biological and genetic characteristics of T. gondii are of utmost importance for the comprehension of the complex host-parasite relationship. Thus the aim of this study was to evaluate the concordance of laboratory techniques used to detect the parasite; to determine the prevalence, biology and molecular epidemiology of T. gondii isolated from stray cats and backyard chickens from the metropolitan region of Goiania, in order to better comprehend the infection. This is the first performed study to determine the genetic and biologic characterization of T. gondii in the state of Goias. This study used 24 stray cats captured by the Center for the Zoonosis Control in Goiania and 50 backyard chickens from the Metropolitan Region of Goiania, Goias. The serologic triage was performed by IHA and showed positivity of 87.4% (21/24) cats and 96% (48/50) backyard chickens for T. gondii. The bioassay was performed using the brain and hearts of the cats and chickens obtained after euthanasia of the animals. After the peptic digestion the homogenate tissues was intraperitoneally inoculated in groups of three Swiss mice each which were daily observed in order to detect signs of acute toxoplasmosis. The asymptomatic mice were euthanized after 60 days followed by serologic analysis through indirect immunofluorescence. Fragments of brain from these animals were observed under optic microscopy in order to visualize tissue cysts. Part of the homogenate was submitted to DNA extraction and polymerase chain reaction (PCR) in which 75% (18/24) cats and 64% (32/50) backyard chickens were positive. The correlation between IHA and PCR from both animals was considered weak. It was not possible to obtain isolate strains from cats. From the chickens it was possible to obtain 15 isolates strains from which 8 presented tachyzoites (acute toxoplasmosis) and 7 presented brain tissues (chronic toxoplasmosis). After the DNA extraction from the isolates the RFLP-PCR was performed using the following primers SAG1, 5’-3’ SAG2, altSAG, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. It was possible to define the genotype from 9 isolates. According to ToxoDB seven isolates corresponded to genotype #65 and two isolates have not been described previously. The genetic analysis showed high diversity and variability. The virulence essays showed that the mortality rate in mice in seven isolates that presented tachyzoites, from the same genotype, detected high virulence in 4 isolates and intermediary virulence in 3. The morphometry analysis of the tachyzoites of these isolates showed statistical difference in at least one of the analyzed variables such as length or nuclei-apical complex distance. These phenotypic differences in isolates from the same genotype show the need to the continuity of the genetic characterization of the parasite using other primers which could be related to the isolated phenotype. The study demonstrated the importance of knowledge about the molecular epidemiology of T. gondii in the metropolitan region of Goiânia, which presents high rates of seroprevalence in hosts.Item Identificação de biomarcadores para o diagnóstico da toxoplasmose aguda e congênita(Universidade Federal de Goiás, 2020-11-27) Storchilo, Heloisa Ribeiro; Castro, Ana Maria de; http://lattes.cnpq.br/9232309971000621; Borges, Clayton Luiz; Alves, Daniella de Sousa Mendes Moreira; Rezende, Hanstter Hallison Alves; Paccez, Juliano Domiraci; Castro, Ana Maria deIntroduction: The diagnosis of congenital toxoplasmosis is complex and might take months to confirm since most newborns do not present symptoms and levels of undetected IgM and IgA anti-Toxoplasma gondii antibodies. Besides, pregnant women may have residual IgM, making it difficult to diagnose acute infection during pregnancy. Despite the various serological tests developed, there are few studies about biomarkers that can assist in the diagnosis of acute and congenital toxoplasmosis. Objective: Analyze the immunoreactivity profile of T. gondii protein with the potential to be biomarkers for the diagnosis of acute toxoplasmosis and the diagnosis and prognosis of congenital toxoplasmosis. Methods: Serum samples were selected from children with symptomatic and asymptomatic congenital toxoplasmosis and women of childbearing age or pregnant women with acute and chronic acquired toxoplasmosis. These samples were grouped according to the patients' laboratory and clinical results. Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE) was used to separate T. gondii proteins. The Immunoblotting technique was applied to the analysis of immunoreactive bands profile by IgG antibodies, using nitrocellulose membranes sensitized with parasite proteins. Results: One hundred and sixteen samples were analyzed. When comparing immunoreactive bands profile for antibodies present in samples from different groups and subgroups, the 150, 18.5, and 16.96 kDa proteins were more immunoreactive by the antibodies present in serum samples from the acquired infection group. The 343, 189, 150, 75, and 42 kDa proteins showed more chance to be detected by the symptomatic congenital infection subgroup, while the 61, 50, and 16.96 kDa proteins were significantly immunoreactive by the acute infection subgroup. Conclusions: Before obtained results, it was possible to identify potential biomarkers that may assist in early diagnosis and treatment of toxoplasmosis, avoiding the appearance of clinical manifestations throughout the life of children congenitally infected.Item Identificação de novos fármacos antimaláricos através de estratégia de quimiogenômica por reposicionamento e validação experimental(Universidade Federal de Goiás, 2018-03-05) Rodrigues, Juliana; Cravo, Pedro Vitor Lemos; http://lattes.cnpq.br/1059199347781390; Andrade, Carolina Horta; Andrade, Carolina Horta; Andrade, Éverton Kort Kamp; Castro, Ana Maria; Neves, Bruno Júnior; Costa, Fabio Trindade MaranhãoMalaria is an infectious disease of possible chronic evolution that affects billions of people in the tropics and subtropics. P. falciparum is the most lethal malaria parasite of humans, while P. vivax is the most widely distributed. The effectiveness of the antimalarial treatment is compromised by the ability of the parasite to evolve resistance to the compounds and by the lack of new effective antimalarials, underscoring the urgent need for the discovery of new drugs. One of the strategies that has been gradually explored in the search for new therapies is the so-called "drug repurposing" approach. In this context, the goal of the present study was to use a drug repurposing-chemogenomics strategy to identify effective drugs against malaria parasites. A comparative genomics tool available from the TDR Targets Database was used through to select targets expected to be present exclusively in P. falciparum and P. vivax parasites, but absent in humans. Each of the selected targets was then used as a query in the following databases: Drugbank, Therapeutic Target Database and STITCH. The P. falciparum and P. vivax targets were aligned with their predicted homologue targets, using pairwise BLAST, to compare functionally relevant regions. Only those where ≥ 80% overlap was observed between the two sequences for the corresponding drug target were considered for subsequent studies. Thereafter, the drugs identified were submitted to a bibliographic search to find drugs that were never evaluated against malaria parasites in the past. A prediction of active compounds was performed through binary QSAR models. The selected drugs were submitted to in vitro assays using asexual stages of P. falciparum (strains 3D7, chloroquine-sensitive, and W2, multidrug-resistant). Epirubicin displayed potent in vitro activity against the 3D7 strain (IC50 = 140 nM). In addition, the drug was shown to be about twice as active against the W2 resistant strain (IC50 = 69 nM), exhibiting even greater activity than chloroquine. At present, in vitro experiments in sexual stages (ookinete conversion) and in vivo assays with P. berghei and P. chabaudi are being carried out. In conclusion, epirubicin is a good antimalarial drug candidate, although future studies are required to investigate its mechanism of action, potential toxicity, and eventually, to advance in the drug development process.Item Avaliação da densidade mineral óssea e estimativa de risco de fraturas em homens vivendo com HIV(Universidade Federal de Goiás, 2018-12-17) Sousa, Clarissa Alencar de; Turchi, Marília Dalva; http://lattes.cnpq.br/3769826743537934; Turchi, Marília Dalva; Siqueira Júnior, João Bosco; Teles, Sheila Araújo; Silva, Nilzio Antonio da; Albuquerque, Maria de Fátima Pessoa Militao deEvaluation of bone mineral density and fracture risk estimation in men living with HIV. Introduction: HIV carriers are at increased risk of developing chronic noncommunicable diseases at an early age. The reduction of bone mineral density, before the age of 50 years, is described in this population. Few studies evaluate the magnitude of osteoporosis in men with HIV, as well as screening strategies in this segment. Objectives: To estimate the prevalence and to investigate risk factors for changes in bone mineral density (BMD); compare osteoporosis prediction model, with bone-densitometry as the gold standard; to evaluate the risk of fractures using an algorithm in HIV-positive men. Method: Cross-sectional study, population of men with HIV, over 40 years old, using ART, attended in Goiânia-Goiás. Participants answered a standardized questionnaire, performed clinical evaluation and laboratory tests. All investigated bone mineral density through dual energy X-ray absorptiometry (DXA). Osteoporosis was defined as bone mineral density less than / equal to 2.5 standard deviations (SD), considering as a reference a young, healthy population categorized by sex. Osteopenia was defined as a reduction between 1 and 2.4 SD of bone mineral density according to WHO criteria. The Osteoporosis Self-Assessment Tool (which uses weight and age data) was evaluated in the prediction of osteoporosis and Low BMD. A ROC curve was constructed to assess OST performance. Results: The participants' age ranged from 40 to 72 years, with a mean of 49.6 (SD = 7.5). The prevalence of low bone mineral density was 56.8% (95% CI: 49.25-64, 13%) and osteoporosis 16% (95% CI 11.0 - 22.1). The risk of fractures in 10-years ranged from 1.1% to 20%, with a median of 1.9%. The factors associated with bone density reduction were the use of tenofovir disproxil fumarate and low BMI. The osteoporosis prediction instrument (OST) presented values of area under the curve of 0.71 and 0.67 in the prediction of osteoporosis and low bone mineral density. The cutoff point 7 obtained the best performance in predicting osteoporosis. The instrument revealed low to moderate predictive power over low bone mineral density and osteoporosis compared to DXA. Conclusion: There was a high prevalence of BMD reduction and overweight in a population of relatively young men with HIV.Item Avaliação bioquímica in vitro do metabolismo energético de cisticercos de Taenia crassiceps expostos a um derivado benzimidazólico, RCB20(Universidade Federal de Goiás, 2015-02-23) Fraga, Carolina Miguel; Castro, Ana Maria de; http://lattes.cnpq.br/9232309971000621; Vinaud, Marina Clare; http://lattes.cnpq.br/1921551651088660; Cruz, Aparecido Divino da; Costa, Tatiane Luiza da; Bezerra, José Clecildo Barreto; Fernandes, Éverton Kort KampHuman infections caused by Taenia spp may be more frequent than they are reported especially if occurred in immunodepressed patients and if the location of the parasites does not involve vital organs. Albendazole is one of the drugs used in the treatment of such infections and its use in mass treatment campaigns may accelerate the resistance development from the parasites both in humans and animal hosts. The aim of this study was to in vitro evaluate the biochemical alterations in T. crassiceps cysticerci after the treatment with a benzimidazolic derivatice (RCB20). The cysticerci were removed from the peritoneal cavity of BALB/c mice with 30 days of infection, macroscopically classified and harvested into 6 well culture plates. 30 cysticerci from each development stage in each well received 5mL of supplemented RPMI culture medium which contained sulfoxide albendazole (ABZSO) (6,5 μM or 13 μM), or RCB20 (6,5 μM or 13 μM). After 24h of culture the cysticerci were removed from the culture medium and both were frozen in liquid nitrogen. Liquid chromatography of high performance and spectrophotometric analysis were performed. It was possible to observe a difficulty in the secretion of substances by the treated cysticerci due to differences in the concentrations of the products dosed from the vesicular fluid and from the culture medium. This was an effect from the mode of action of the drugs which affects the tubulin formation. Regarding the glycolysis it was possible to observe a gradation increase in the secretion/excretion (S/E) of lactate in the treated groups while there was a decrease of this organic acid in the vesicular fluid. As to the energetic metabolism and the citric acid cycle it was possible to observe its traditional sense functioning due to the detection of alf- ketoglutarate. It was possible an increase in the concentrations of citrate, malate, fumarate and succinate in the treated groups. The alternative pathway of energy production such as beta-oxidation of fatty acids and protein catabolism were detected with an increase in the metabolites from those pathways in the treated groups. Therefore we conclude that in spite of not blocking the glucose uptake the drugs influenced its uptake which lead to the use of alternative pathways of energy production. In spite of more soluble and stable the RCB20 formulation did not present a different biochemical effect than ABZSO.Item Avaliação da capacidade da nova linhagem de células dendríticas AP284 responder a diferentes agonistas de TLRs e de induzir uma resposta Th17(Universidade Federal de Goiás, 2018-11-14) Oliveira, Pollyana Guimarães de; Oliveira, Milton Adriano Pelli de; http://lattes.cnpq.br/2152513705182408; Oliveira, Milton Adriano Pelli de; Fonseca, Simone; Pfrimer, Irmtraut Araci Hoffmann; Campos, Helioswilton Sales de; Lacerda, Elisângela SilveiraDendritic cells (DCs) are antigen presenting cells (APCs) that belongs a diversify group of cells, being still the main cells that can presents the antigen and activate T lymphocytes by inducing different kinds of cytokine production, which can influence the development of different profiles of lymphocytes. The production of IL-12p70 leads to the Th1 profile differentiation, furthermore the IL-23 production can be important for the establishment and maintenance of Th17 profile. Thus, IL-12 and IL-23 play an important role in the induction or establishment of the adaptive immune response. The aim of the study was characterize the DC AP284 cell line by checking its surface markers, TLRs expression and capacity of response to different TLRs agonists. Besides of that, the production of IL-12p40, IL-12p70, IL-23 and the ability to induce Th1 or Th17 profile were evaluated. AP284 cells have expressed surface molecules compatible with DCs including class II MHC, CD11c and 33D1. After stimulation using different TLRs agonists these cells produced higher amounts of de IL-12p40 and IL-23, but no IL-12p70 production was observed. The IFN-γ addition was not able to induce IL-12p70 production, instead of that an inhibition of the IL-12p40 and IL-23 production were observed. The results have also shown that AP284 cell line is related with the Th17 profile because higher amounts of IL-12p40 and IL-23, but not IL-12p70 were observed, besides of that the results showed an increase in IL-17 levels produced by mice lymph node cells after immunization with Freund’s adjuvant and BCG. Thus, AP284 cells can be important to study the mechanisms of induction and regulation related with the production of IL-12p40 and IL-23 working as a possible tool to study the establishment and maintenance of Th17 cells profile. Moreover, AP284 cells have some similar characteristics among the other immortalized murine DCs, but the AP284 line produces a much higher amount of IL-12p40 than all described cells and is also differentiated among them by the production of IL-23.Item Influência da inflamação crônica de baixo grau na susceptibilidade à tuberculose e resposta vacinal em camundongos(Universidade Federal de Goiás, 2019-10-14) Resende, Danilo Pires de; Kipnis, André; Kipnis, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; Kipnis, Ana Paula Junqueira; Fonseca, Simone Gonçalves da; Cominetti, Cristiane; André, Maria Cláudia Dantas Porfírio Borges; Campos, Helioswilton Sales deChronic conditions associated with inflammation, such as obesity (OB), diabetes mellitus (DM) and tuberculosis (TB), are serious public health problems. The OB epidemic is increasing worldwide; and the prevalence of adult obesity nearly doubled between 1980 and 2017. Since DM is associated with diabetes, the prevalence of diabetes also increased over the same period, reaching 9% of the adult population worldwide, totaling 422 million people. Studies have been demonstrating DM as a risk factor for the development of tuberculosis, raising the hypothesis of an association between OB, DM and TB. Circulating monocyte subpopulation proportions have been shown to change during OB, DM, and TB, however no studies have evaluated monocyte subpopulations in severe OB / DM and if there are characteristics that may be associated with increased susceptibility to TB. In the first part of this study we demonstrated, in a cross-sectional study, with individuals with severe OB (BMI over 35kg / m2) with or without OB (DM) (n = 50 individuals per group) that the population did not of monocytes from OBDM individuals presents similar characteristics to monocytes from individuals with active pulmonary TB, and monocytes from OBDM and TB individuals are more susceptible to Mycobacterium tuberculosis (Mtb) infection. This suggests that phenotypic similarity and susceptibility of monocytes may be factors contributing to the association between tuberculosis and obesity. Since monocytes/macrophages change in obesity and these changes interfere with the ability to control Mtb, we decided to assess whether the development of obesity could alter the ability to rescue the specific immune response following M. bovis BCG vaccination. Thus, in the second part of this paper, we evaluated whether obesity interferes with BCG vaccine response by vaccinating young C57BL / 6 mice and inducing obesity by hypercaloric diet and by analyzing the immune response after Mtb challenge. Obesity induced after BCG vaccination did not interfere with TCD4 + IFNγ + lymphocyte rescue after Mtb challenge. However, this response to Mtb has been reduced. These results suggest that vaccination of non-obese animals induces vaccine immune responses that do not change with obesity induced metabolic changes.Item Avaliação de receptores similares a toll (tlrs) em monócitos de pacientes com doença de Parkinson(Universidade Federal de Goiás, 2017-06-02) Rocha Sobrinho, Hermínio Maurício da; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Dias, Fátima Ribeiro; Silva, Marcos Vinícius da; Cruvinel, Wilson de Melo; Diniz, Denise Sisterolli; Gomes, Rodrigo SaarParkinson's disease (PD) is a condition of the central nervous system (CNS), chronic and progressive, resulting from the death of brain dopamine-producing neurons. Genetic and epidemiological studies have highlighted the role of neuroinflammation in the pathophysiology of PD, emphasizing that the process of neuroinflammation is associated with the death of dopaminergic neurons. In peripheral blood or tissues, leukocytes activated by pathogen-associated molecular patterns (PAMP) and / or tissue damage (DAMP), via Toll-like receptors (TLR), produce pro and antiinflammatory mediators whose imbalance can lead to a chronic inflammation. Under physiological conditions, three subpopulations of human peripheral blood monocytes are detected: the classical monocytes (CD14++CD16-), the intermediates (CD14++CD16++) and the non-classical (CD14+CD16+), these cells present phenotypic and functional differences that can influence the characteristics of the immune response against DAMPs or PAMPs. The aim of this study was to compare the peripheral blood leukocyte potential of patients with PD or healthy individuals in the production of cytokines (TNFα and IL-10) after stimulation of these cells with TLR4 (LPS) or TLR2 / 1 (Pam3Cys) agonists, to evaluate the effect of the age of DP patients on the production of cytokines in blood cultures, to investigate the percentage of monocyte subpopulations expressing TLR1, TLR2 and TLR10, to evaluate the influence of TLR10 expression on the biological activity of TLR2 to its agonist in PBMC cultures. Two groups of patients with PD were evaluated (≤ 55 years old and ≥ 65 years old), age-matched with healthy controls (n = 26). 6 h-TNFα production was increased after TLR4 stimulation, mainly in young PD patients, whereas TLR2-induced TNFα and IL-10 levels were decreased in PD patients independent of age (p < 0.05). A reverse correlation between LPS-induced TNFα production and age was observed in PD patients and controls, but TNFα induced by TLR2 agonist was not associated with age of PD patients or controls. TNFα production induced by TLR4 but not by TLR2 was reversely associated with the age at PD onset and disease duration. No associations between UPDRS scores and cytokine levels were detected. A higher percentage of CD14++CD16+ monocyte subpopulation was observed in the blood of patients with PD compared to healthy controls, as well as a higher percentage of monocytes from patients expressing TLR10. The expression of TLR2 and TLR10 in peripheral blood monocytes from PD patients was increased relative to that of healthy controls. The results suggest that the peripheral blood leukocyte response to the TLR2 ligand is reduced in PD and that the elevation of TLR10 expression in monocytes influences inhibiting the biological activity of TLR2 to its agonist as demonstrated in the neutralization experiment of TLR10. In this study, the group of DP patients with the highest severity of the disease, evaluated by the UPDRS scale, had a lower percentage of monocytes expressing TLR10, suggesting the expression of this receptor in peripheral blood leukocytes as a possible prognostic marker for DP. These findings, together, may play an important role in the immunopathology of PD. The study of TLR cell expression in peripheral leukocytes and CNS, intracellular signaling pathways and the production of pro and anti-inflammatory mediators is necessary in order to better clarify the role of TLR in the PD pathogenesis and perhaps for the identification of biomarkers of disease prognosis in the blood PD patients.Item Acidentes de trânsito em capitais selecionadas do Brasil: estimativa da magnitude corrigida e fatores associados à gravidade da lesão(Universidade Federal de Goiás, 2017-08-24) Mandacarú, Polyana Maria Pimenta; Morais Neto, Otaliba Libânio de; http://lattes.cnpq.br/4030124246791320; Morais Neto, Otaliba Libânio de; Duarte, Elisabeth Carmen; Mota, Eduardo Luiz Andrade; Ternes, Yves Mauro Fernandes; Itria, AlexanderIntroduction: Middle-and low-income countries currently account for 92% of all road transport fatalities worldwide, with an increasing trend in mortality rates, the opposite of what occurs in high-income countries. Brazil has a high morbidity and mortality burden caused by traffic. However, one of the limitations of the knowledge of the real magnitude of traffic accidents in Brazil is the lack of qualified information about traffic accidents by mode of transportation and the underestimation of the actual number of fatalities and serious injuries. In this way, the qualification of the databases through the relationship of health and traffic records allows improving coverage, coverage and quality of information, as well as enhances the epidemiological analysis of this disease in the population. Objectives: To estimate the magnitude of deaths and severe injuries using a linkage procedure as well as the percentage of correction for health and traffic data sources in the municipalities of Belo Horizonte, Campo Grande, Curitiba, Palmas, Teresina and Goiania, and to characterize the factors Associated with deaths and serious injuries in Goiania. Method: Two cross-sectional studies were conducted, using a database of traffic victims (VIT), the Hospital Inpatient System (SIH), and the Mortality Information System (SIM). The first in Belo Horizonte, Campo Grande, Curitiba, Palmas and Teresina and the second in Goiania. A linkage procedure was performed in both studies through the RECLINK III program, identifying true pairs with calculation of the percentage of correction of the underlying cause of death, secondary diagnosis or classification of the victim in the traffic database. In the second study, for the definition of the associated factors for deaths and severe injuries, the incidence ratios with a 95% confidence interval were estimated. The comparison of the incidences between the categories of each variable using bivariate and multivariable regression model using the Poisson regression, with robust variance. Results: The results showed that there was a considerable correction of the basic cause of death, diagnosis of hospitalization or classification of the severity of the victim's injury in traffic records in the six capitals. For SIM, the percentage of correction of the underlying cause of death was 29.9%, 11.9%, 4.2%, 33.5%, and 43.9% for Belo Horizonte, Campo Grande, Curitiba, Teresina and Goiania, respectively. For SIH, the percentage of correction of the secondary diagnosis of hospitalization was 51.3% for Goiania, 24.4% for Belo Horizonte, 96.9% for Campo Grande, 100.0% for Palmas and 33.2% for Teresina. For VIT, there was a change in the classification of the severity of the victim (not severe to severe), with correction percentage of 100.0% for Belo Horizonte and Teresina, 48.0% for Campo Grande, 52.8% for Goiania and 51.4% For Palmas. In the case of nonfatal to fatal, the correction was 29.5%, 52.3%, 74.3%, 4.4% and 72.9%, respectively, for Belo Horizonte, Campo Grande, Curitiba, Palmas and Teresina. For Goiania, the contribution of the linkage procedure to the database of victims was the identification of 15 deaths (9.6%), not classified as such in the transit data base. In Goiania, 70% of all victims were males and 43.7% of all victims were aged between 18 and 29 years and 63% of all accidents were motorcycle occupants. The main factors associated with death were: age over 40 years (40-49 years: RI 2.75, IC 1.11-6.79, 50-59 years: RI 4.46, IC 1.8- 11.04 and 60 and more: RI 7.69, IC 3.15-18-78) bicycle occupants (RI 2.26 IC 1.19-4.3) and pedestrians (RI 2.12 IC 1.26 -3.58) and the occurrence of the accident between 0-6 hours (RI 2.47 IC 1.36-4.47); For the severely injured were: the age group over 40 years (40-49 years: RI 1.62, IC 1.26-2.08, 50-59 years: RI 1.48, IC 1.23-2, 16 and 60 and more: RI 2.00, IC 1.50-2.66), occupants of Motorcycle (RI 2.38 IC 2.01-2.83), Bicycle (RI 2.35 IC 1.76- And the occurrence of the accident between the periods of 00: 00-17: 59 hours (00:00 to 05:59 RI 1, 38 IC 1.1-1.73.06.06 at 11.59 RI 0.72 IC 0.63-0.83; 12:00 at 17.59 RI 0.84 IC 0.73-0.95). Conclusion: The study contributed to the qualification of the coverage and quality of the information of the health and traffic data banks, as well as identified gaps and limitations in the information system that registers ATT.Item Genomas completos e parciais de formas recombinantes BF1 e BC do HIV-1 circulantes nos estados de Goiás, Mato Grosso, Mato Grosso do Sul, Tocantins, Maranhão e Piauí(Universidade Federal de Goiás, 2017-08-14) Reis, Mônica Nogueira da Guarda; Guimarães, Monick Lindenmeyer; http://lattes.cnpq.br/0776265935831372; Stefani, Mariane Martins de Araújo; http://lattes.cnpq.br/5581414958714905; Stefani, Mariane Martins de Araújo; Santos, André Felipe Andrade; Delatorre, Edson Oliveira; Araújo Filho, João Alves de; Cardoso, Ludimila de Paula VazBackground and objectives: One of the most striking features of HIV-1 is its extensive genetic polymorphism which allows its classification into four groups (M, N, O and P). The pandemic group M can be classified into nine subtypes (A-D, F-H, J, K), six sub-subtypes (A1-A4, F1-F2), dozens of circulating recombinant forms (CRFs) and countless unique recombinant forms. The molecular epidemiology of HIV-1 in Brazil is complex and dynamic and has been characterized by the cocirculation of subtypes B, F1 and C and BF, BC, BFC and CF recombinants. Previous studies on pol sequences of HIV-1 isolates from six Brazilian States (Goiás, Mato Grosso, Mato Grosso do Sul, Tocantins, Maranhão e Piauí) have shown great variability of BF, BC and FC recombinant viruses. This thesis describes the molecular characterization of these mosaic genomes including reclassification in pol and generation of full length/near full length/ partial genomes of representative isolates. The results of this thesis are presented as articles, one of them is already published and two other are presented as manuscripts to be submitted. Methods: Proviral DNA was extracted from whole blood and four overlapping fragments that compose HIV-1 whole genome were amplified by “nested”-PCR. The generated sequences were aligned (BioEdit 7.2.0) and phylogenetic analysis performed (MEGA 6, Neighbor-Joining, Kimura 2 parameters). The recombination profiles were identified by point analysis, phylogenetic analysis of fragments, and Bootscan analysis (SIMPLOT v3.5.1). The time of the most recent common ancestor (TMRCA) of HIV-1 BF clades was estimated (Bayesian Markov Chain Monte Carlo, BEAST v1.8). Results: Article 1/Plos One: Among 828 HIV-1 isolates from six Brazilian States, phylogenetic analysis of pol identified 87 BF recombinant isolates: 48% (42/87) grouped into five different clusters (Cluster #1-5), 21% (18/87) was CRF_BF-like and 31% (27/87) were classified as URFs_BF. Among 22 isolates that composed the largest BF cluster (#5), we have obtained six full length genomes, one near full length genome and four partial genomes. These 11 isolates, obtained from patients that did not have any epidemiological link shared identical recombination profile in their genomes, allowing the description of a new CRF. This recombinant named CRF90_BF1 is circulating in Goiás, Mato Grosso e Tocantins states. Among 20 BF isolates from the other four clusters we have obtained: three full length genomes, four near full length genomes and five partial genomes. Analysis of these sequences characterized three URFs (Clusters #1, #2 e #3), circulating in the Central West region. The estimated TMRCA for these BF clades was the beginning of the 90’s. BLAST search analysis identified other sequences sharing the same recombination pattern among isolates from the North and South regions, suggesting that they may represent other potential, unidentified CRFs. A moderate rate of CRF28/CRF29_BF-like isolates was seen (16.1%, 14/87) in Goiás, Mato Grosso and Mato Grosso do Sul states. Manuscripts 1 and 2: BC and BF recombinant isolates were also detected (2.9%; 24/828): BC (2,3%, 19/828), BFC (0,4%, 3/828) and CF (0,2%, 2/828). Among these 24 isolates, 19 grouped into six clusters while five (BRGO4156, BRMT2509, BRMT3086, BRMS43, BRPI34) did not cluster. Six clusters were identified (Clusters #1-6): two clusters contained 7 isolates with recombination profiles similar to already identified CRFs_BC (29.2%, 7/24). Cluster #4/CRF31_BC-like comprised 5 isolates (20.8%) and two isolates were Cluster #5/CRF60_BC-like (8.3%). Most patients infected with BC and FC isolates was antirretroviral naïve and (23/24) and 21.7% (5/23) had mutations associated with transmitted drug resistance. Conclusions: Our analyses of isolates from Central West, North and Northeast regions allowed the characterization of the new CRF90_BF1 and different URFs_BF and BC from already described ones. Also isolates sharing recombination profiles with already described ones were also observed (CRF28/29_BF-like, CRF31_BC-like, CRF60_BC-like). Altogether our results indicate that coinfection/ superinfection and intersubtype recombination may be more prevalente that reported. Also, our results indicat the continuing generation of recombinants in the Brazilian epidemic that is taking place in urban centers located away from the epicenter of the epidemic. Continued surveillance studies including full/near full genomes in larger sample sizes are necessary to identify emerging and disseminating recombinants and their implication in the transmission and control of HIV-1 in Brazil.Item Identificação de novos compostos bioativos contra tripanossomatídeos a partir de estratégias integradas em química medicinal(Universidade Federal de Goiás, 2018-04-02) Melo Filho, Cleber Camilo de; Braga, Rodolpho de Campos; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4731114E4; Andrade, Carolina Horta; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4745602P1; Andrade, Carolina Horta; Castro, Ana Maria de; Oliveira, Milton Adriano Pelli de; Lacerda, Elisângela de Paula Silveira; Scotti, Marcus TulliusTrypanosomatids are parasites that cause different diseases affecting mainly low-income populations from countries in development or underdeveloped countries. These diseases are classified as neglected tropical diseases (NTDs). The few drugs available for the treatment of these parasitic diseases present several problems regarding low efficacy, resistance, and toxicity, making the need of new therapeutic options an urgent task. In this sense, the aim of this work was the identification of novel drug candidates against trypanosomatids through the application of integrated strategies in Medicinal Chemistry. To achieve this goal, three studies were performed combining different computer assisted drug design (CADD) approaches and experimental validation. In the first study, 39 virtual hits were identified by quantitative structure activity relationships (QSAR)-based virtual screening (VS) and selected for experimental evaluation. Among them, 13 compounds were active and selective in vitro against intracellular T. cruzi. Additionally, an in silico multi-parameter analysis for evaluation and comparison of some pharmacokinetic (ADMET) and physicochemical properties, with potency and selectivity, allowed the prioritization of the most promising compounds for further in vivo assays. In the second study, VS was performed for identification of novel potential inhibitors of pyruvate kinase (PK) from Leishmania spp., allowing the discovery of 14 novel potential inhibitors of this enzyme. These compounds were submitted to experimental evaluation against L. infantum amastigotes. In the third study, a dual-target VS was performed, allowing the identification of 15 potential inhibitors of both PK and sterol 14 α-demethylase (CYP51) of Leishmania spp., also predicted as active compounds against L. infantum amastigotes by QSAR models. Therefore, this work has demonstrated the potential of the integration of computational and experimental methods for the identification of active compounds against trypanosomatids.Item Relação entre a presença de macrófagos no microambiente tumoral e fatores prognósticos em câncer de mama(Universidade Federal de Goiás, 2018-04-26) Labre, Luciana Vieira Queiroz; Saddi, Vera Aparecida; http://lattes.cnpq.br/7496804650895441; Santos, Silvia Helena Rabelo dos; http://lattes.cnpq.br/4994826511439492; Santos, Silvia Helena Rabelo; Freitas Júnior, Ruffo de; Batista, Aline Carvalho; Celes, Mara Rúbia Nunes; Costa, Nádia do LagoIntroduction: The analysis of possible biomarkers associated with the more aggressive behavior of breast carcinomas, linked to clinical and prognostic information, can provide important data in the treatment of patients with breast cancer. There are indications that the presence of tumor associated macrophages (TAMs) in the tumor microenvironment is associated with a worse prognosis for women with breast cancer. Objective: To evaluate the importance of the presence of macrophages in the tumor microenvironment in breast neoplasms. Methods: This retrospective cross-sectional study included 93 cases of women with invasive ductal breast cancer and submitted to immunohistochemical analysis of the markers for CD68 (M1 / M2) and CD163 (M2) macrophages. The counts of CD68 + and CD163 + cells were performed in a peritumoral region, considering hot spot fields, and the cases under study were classified as low infiltration or high infiltration using semiquantitative evaluation of cell marking. Statistical analysis was performed using the Stata 13.0 program (Stata Corp., College Station, TX, United States), with bivariate analysis, considering the level of significance of p <0.05, and multivariate analysis where the variables with p <0.2 were submitted to the logistic regression test. The Kaplan-Meier method and the log-rank test were used in the analysis of survival curves. Results: High levels of TAMs (CD68 +) were significantly correlated with: patient age, presence of distant metastasis, more advanced tumor staging, absence of estrogen receptors (ER) and progesterone (PR), and triple negative breast cancer (TN). The reduction of the overall survival rate in the cases of women with breast cancer studied was significantly associated with high infiltration of TAMs (CD68 +). The high infiltration of TAMs phenotype M2 (CD163 +) was significantly correlated with the independent variables presence of distant metastasis, absence of RE and TN breast cancer. Conclusions: The association of the analysis of TAM biomarkers with clinical and prognostic information showed that the high presence of macrophagic infiltrate has a significant prognostic value, adding information on the relationship between the presence of macrophages in the tumor microenvironment of cases of women with carcinoma and the clinical-pathological characteristics of the tumor. In addition, TAMs appear as potential prognostic markers for breast cancer.Item Estudo a homeostase de cobre no fungo patogênico histoplasma capsulatum(Universidade Federal de Goiás, 2018-07-18) Tristão, Gabriel Brum; Bailão, Alexandre Melo; http://lattes.cnpq.br/5415221996976886; Bailão, Alexandre Melo; Malavazi, Iran; Lima, Patrícia de Sousa; Bailão, Elisa Flávia Luiz Cardoso; Paccez, Juliano DomiraciHistoplasma capsulatum is a thermodymorphic pathogenic fungus that causes systemic mycosis known as histoplasmosis. This fungus grows as mycelium at temperatures around 25°C and as yeast at 37°C. During the infectious process, pathogenic microorganisms must obtain nutrients from the host in order to survive in infected tissues. Among these nutrients, copper is an essential metal ion, because it participates in oxidation/reduction reactions, in energy production, in the transport of electrons, its cofactor of many enzymes and metalloproteins and is required for energy and melanin production. Copper excess however, it is toxic due to the fact that produces reactive oxygen species, dislocates other metals from metalloproteins, causes damage to lipids and DNA, so because of this H. capsulatum must maintain the homeostasis of this metal during infection. We observed here that H. capsulatum, through the transcriptional levels of Ctr4, Mac1, Crp1 and Ace1, during the infectious process in macrophages, faces an environment of copper overload imposed by the host cells via copper ATPase ATP7a. This copper excess shown to be INF-ɣ and time dependent, because when macrophages are not stimulated by INF-ɣ and in greater times of infection, they impose a restrictive copper environment instead, during the infection. H. capsulatum uses the Crp1 copper efflux pump in order to respond this toxic copper milieu, since mutant yeasts for Crp1 were unable to grow at high levels of the metal. Despite the importance of Crp1 for the fungus in this context, it appears that Crp1 is not strictly necessary for the total virulence of the fungus, leading us to infer that other proteins may also be exerting a Crp1-like function in H. capsulatum. It is clear once again that in the pathogen-host relationship copper plays a complex, highly dynamic and dependent on certain biological variables role.Item Detecção e genotipagem do papilomavirus humano em amostra do colo do útero de adolescentes do Distrito Sanitário Noroeste do município de Goiânia, Goiás(Universidade Federal de Goiás, 2005-03-31) Daud, Lyana Elias Santos; Alves, Maria de Fátima Costa; http://lattes.cnpq.br/1412047788155346; Alves, Maria de Fátima Costa; Martins, Regina Maria Bringel; Villa, Luísa LinaBackground: Human Papillomavirus (HPV) infection is a strong predictor for cervical cancer development. Some HPV types are considered at high-risk and associated with pre-malignant lesions and cervical cancer. Genital HPV infection is highly prevalent in sexually active young women. Actually PGMY09/11 primer system associated with Restriction Fragment Length Polymorphism (RFLP), hibridization or DNA sequencing is one of the most sensitive methods for detecting and typing HPV-DNA in genital samples. Objectives: To detect and identify HPV-DNA genotypes in genital specimens among sexually active female adolescents from Distrito Sanitário Noroeste, Goiânia. Methods: We performed a cross-sectional study among 432 sexually active female adolescents (15-19 years), randomly selected at Distrito Sanitário Noroeste, Goiânia, and served by the Family Health Program. HPV-DNA was detected by PCR assay, in endocervical samples, using the PGMY09/11 consensus primer set. The presence of amplifiable DNA was assessed with -globin primers (PCO3/PCO4). Amplicons generated with PGMY primers were typed with the line blot assay (PGMY-line blot) and RFLP. Results: HPV-DNA was detected in 121 samples yielding a 28% prevalence (95% CI 23.8- 32.5). All samples yielded a -globin amplimer, confirming the DNA adequacy. Thirty different HPV genotypes were identified, the high risk genotypes frequently found being: HPV-16 (6,7%); followed by HPV-51 (5,1%); HPV-31 (4,6%); HPV-52 (4,2%) and HPV-18 (3,5%). PGMY-line blot test identified 54 cases of infection caused by multiple genotypes. The genotypes 16, 51, 18, 53 and 52 were the most frequently associated with cervical co- infection with multiple HPV types. Conclusions: A high prevalence of HPV-DNA was found and a broad spectrum of HPV genotypes was identified, predominantly high risk HPV genotypes. A high frequency of multiple HPV infections was found in the studied population.Item Prevalência e caracterização molecular dos vírus linfotrópicos de células T humanas 1 e 2 em primodoadores de sangue e seus familiares no estado do Piauí(Universidade Federal de Goiás, 2014-12-15) Ribeiro, Ivonizete Pires; Pinheiro, Gevina da Silva; http://lattes.cnpq.br/6309974608158589; Martins, Regina Maria Bringel; http://lattes.cnpq.br/2582896795892370; Martins, Regina Maria Bringel; Kozlowski, Aline Garcia; Fiaccadori, Fabiola de Souza; Carneiro , Megmar Aparecida dos Santos; Ferreira, Renata CarneiroHuman T-lymphotropic virus type (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATL), HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), and other inflammatory diseases. HTLV-2 has been associated with a syndrome similar to HAM/TSP and other clinical manifestations. The present study aimed to investigate the seroepidemiologic and molecular aspects of HTLV-1/2 infection in the first-time blood donors and their family members in the state of Piauí. Between August 2011 and July 2012, 37,306 first-time blood donors were screened by enzyme-linked immunosorbent assay (ELISA) for the presence of anti-HTLV-1/2 antibodies in the Centro de Hematologia e Hemoterapia do Estado do Piauí (HEMOPI). The positive individuals were recruited for a second blood collection and interview, when their family members were invited to participate to this investigation. Sera were tested by ELISA, and the positivity confirmed by line immunoassay (LIA). Whole-blood samples of anti-HTLV seropositive individuals were submitted to proviral DNA detection by polymerase chain reaction (PCR) and nucleotide sequencing, followed by phylogenetic analysis. Of 37,306 first-time blood donors, 48 were anti-HTLV-1/2 reactive by ELISA. By testing the second serum samples, 47 (0.13%) were also reactive by ELISA. Among these, 22 (0.06%) were anti-HTLV-1 and 14 (0.04%) anti-HTLV-2 positive by LIA. Most of HTLV-1 or 2 infected patients reported risk characteristics related to vertical and sexual transmission. Genetic characterization demonstrated that all 22 HTLV-1 isolates belonged to the Cosmopolitan subtype HTLV-1a Transcontinental subgroup A. For HTLV-2, 11/14 (78.6%) anti-HTLV-2 positive samples were amplified and then identified as subtype a (HTLV-2 a/c). Of 21 families invited to take part in the study, five agreed to participate. Three had, in addition to the index case (blood donor), two or more anti-HTLV-1 or 2-positive members (ELISA/LIA). By combining epidemiological data of the first family members with molecular analysis of their isolates, intrafamilial/vertical transmission of HTLV-1 was suggested, as well as the transmission of HTLV-2 among the second and the third families. Additionally, HTLV-2 sexual transmission could be considered in the last family. The prevalence of 0.13% (CI 95%: 0.11-0.17) for HTLV-1/2 found in this study is similar to those observed in blood donors in Brazil, and HTLV-1 and 2 subtypes identified in this investigation were consistent with those circulating in Brazil. In addition, this study provides evidence of intrafamilial transmission of HTLV-1 and 2.Item Circulação dos vírus dengue no estado de Goiás: vigilância laboratorial (1994-2013) e perfil de anticorpos neutralizantes sorotipo específico durante o surto de 2013 em Goiânia(Universidade Federal de Goiás, 2014-12-19) Argolo, Angela Ferreira Lopes de Teive e; Féres, Valéria Christina de Rezende; http://lattes.cnpq.br/8089054699896454; Martelli, Celina Maria Turchi; http://lattes.cnpq.br/5867052489026059; Martelli, Celina Maria Turchi; Braga, Maria Cynthia; Cordeiro, Marli Tenório; Turchi, Marília Dalva; Santos, Silvia Helena Rabelo dosDengue is an endemic disease in Goias state with current circulation of the four serotypes. In the last five years has been occurred a significant increase in cases, hospitalizations and deaths caused by dengue in the region. In this thesis the first manuscript analyzed data from the virological surveillance of DENV in Goias during 1994-2013. Virus isolation in C6/36 (A.albopictus) cell culture followed by indirect immunofluorescence using monoclonal antibodies were performed in LACEN-GO. During the study period the four serotypes have been identified with introduction of DENV-1 in 1994, DENV-2(1998), DENV-3(2002) and DENV-4(2011). Among 21,684 virus isolation tests performed 4,839(22.3%) were positive. DENV-1 was isolated in 3,023(62.5%), DENV-3 in 1,051 (21.7%), DENV-4 in 413(8.5%) and DENV-2 in 352(7.3%) samples respectively. There were changes in the predominance of serotypes throughout the study period with dominance of DENV-1 between 1994-2002 and 2009-2011, DENV-3 in 2003-2008 and DENV-4 in 2013. The DENV-2 was never prevalent among serotypes isolated in LACEN-GO. The time series of the virological surveillance of dengue in Goias showed the introduction and the temporal sequence of circulation of DENV in the state. This study contributes to characterize the epidemiological scenario of dengue in Goias with possible implications for epidemiological surveillance of the disease in the region. The second manuscript evaluated the presence of dengue serotype-specific neutralizing antibodies (NAb) in patients involved in a clinical dengue cohort established during epidemic of 2012-2013 in Goiania, state of Goias. Among 452 participants of the cohort we analyzed the paired samples from 60 patients classified as severe dengue (n=40) or dengue fever (n=20). The eligibility criteria were confirmation of the infection by IgM, NS1Ag and /or RT-PCR; IgG positive in at least one of the samples. Monotypic or multitypic response were defined: PRNT50≥1/20 for only one serotype or ≥1/20 for two or more serotypes simultaneously. The infecting serotype was defined by ≥4 fold increase in the title of NAb in paired samples. Overall 73.3% of patients had NAb against DENV-4, 68.3% against DENV-1, 68.3% against DENV-2 or 61.6% against DENV-3. The patients' ages ranged from 3 to 81 years. Regardless of age group 85% of patients had multitypic response while 15% had monotypic response. Patients infected with DENV-4 showed the greatest difference in NAb titers in comparison to other serotypes indicating predominance of seroconversion for serotype 4. There was no correlation among preexistent NAb and disease severity. The characterization of dengue serotype-specific immune response is important to study the relationship of preexistence of neutralizing antibodies with clinical outcomes of disease also facing the perspective of introducing of antidengue vaccines to identify protective immunity.