Mestrado em Biologia da Relação Parasito-Hospedeiro (IPTSP)
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Item Detecção, tipificação e filogenia molecular de Papilomavírus bovino em bovinos leiteiros(Universidade Federal de Goiás, 2017-03-31) Albuquerque, Winnie Castro Amorim e; Braga, Carla Afonso Da Silva Bitencourt; http://lattes.cnpq.br/7673897995590123; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Carneiro, Lilian Carla; Barbosa, Mônica Santiago; Santos, Mônica Oliveira; Moraes Filho, Aroldo Vieira de; Souza, Guilherme Rocha Lino deBovine papillomavirus is the etiological agent of bovine papillomatosis, a disease that triggers warts throughout the skin, udder, roofs, genitalia and in more severe cases can develop extensive papillomas, cause neoplasia in the digestive tract and bladder, weaken the animal's health and cause losses in the Productivity and losses for livestock. The present study aims to detect and typify bovine Papillomavirus present in bovine tissue and blood samples with papillomatosis, to sequence the isolated viral types, to analyze the nucleotide sequences and the phylogeny of the detected viral types. As a result, amplification was obtained in five tissue samples (papilloma) from different bovines, not being successful in the amplification of blood samples. PCR reactions revealed the presence of BPV-1 in 60%, BPV-5 in 40%, BPV-9, BPV-10, BPV-13 and BPV-14 in 20% and BPV-12 in 40% of the analyzed samples. The presence of coinfection was verified in 60% of the lesions analyzed, with up to four viral types infecting the same sample. Alignments of viral type sequences 1, 5 and 14 were validated with identity ranging from 74% to 95%. The phylogenetic diagram showed a genetic approximation between viral types 1 and 14, both belonging to the genus Deltapapillomavirus, and distancing between nucleotide sequences of viral types 5, 9 and 14. Papillomaviruses of types 5 and 9 belong to different genera, Epsilonpapillomavirus and Xipapillomavirus, Respectively, the phylogenetic distance between these viral types, verified in the diagram, is justified.Item Haemophilus influenzae e Haemophilus haemolyticus isolados de crianças que frequentam creches no município de Goiânia-GO: prevalência, fatores de risco e caracterização molecular da resistência antimicrobiana(Universidade Federal de Goiás, 2017-06-26) Almeida, Robmary Matias de; Cardoso, Juliana Lamaro; http://lattes.cnpq.br/0768752229180519; Cardoso, Juliana Lamaro; Kipinis, André; Moraes, Camile deHaemophilus influenzae (Hi) and Haemophilus haemolyticus (Hhae) are important microorganisms present in human nasopharyngeal colonization, with rates varying according to locality, sampling frequency, individual and social factors. Hi is a pathological agent that causes diseases such as meningitis, pneumonia, sepsis and otitis media, which presents in encapsulated forms with six serotypes a, b, c, d, e, f, and uncapsulated or non-typeable (HiNT). Hhae is a nasopharyngeal comensal and rarely causes invasive diseases. The objective of this study was to estimate the prevalence of Hi and Hhae in children under five years of age attending public day care centers in the city of Goiânia-GO, to determine the circulating serotypes, to analyze the risk factors associated with the nasopharyngeal carrige, as well as to characterize the antimicrobial resistance of Hi. Were analyzed 1.188 nasopharynx swabs from healthy children between 36 and 59 months of age from October to December 2010. The samples were submitted to bacterial culture for the isolation of Haemophilus spp. For the identification of the species, the Real-Time Polymerase Chain Reaction (TR-PCR) was used. Serotyping, as well as detection of the bla TEM-1 and bla ROB-1 resistance genes, was performed through the conventional Polymerase Chain Reaction. Phenotypic detection for β-lactamase production was performed by the chromogenic cephalosporin test. The database was constructed with the statistical software SPSS (Chicago, IL, USA) version 18.0. Risk factors, children aged 3 years, low maternal schooling and three or more children under 10 years of age living in the same household of the child recruited in the study were evaluated by multivariate Poisson regression. The prevalence of Hi carriers was 54.4% (646 / 1.188), 0.9% (n = 11) of the serotype e, 0.9% (n = 11) of serotype f, 0.2% (n = 2) serotype a, 0.08% (n = 1) serotype d, 0.0% (n = 0) serotype b and c and 52.3% (621 / 1.188) of HiNT. The prevalence of Hhae was 1.2% (14 / 1.188). Among the encapsulated Hi, the prevalence of the bla TEM-1 gene was 4.0% (1/25) and the bla ROB-1 gene was 4.0% (1/25). Among the 20% (124/621) of HiNT analysed, the prevalence of the bla TEM-1 gene was 13,7% (17/124) and the prevalence of the bla TEM-1 gene was 1,6% (2/124). Continuous surveillance of Haemophilus spp. as a colonizer, is necessary to evaluate its transmission and dissemination in the population where there is a higher risk of invasive disease, to control Hib re-emergence after the vaccinacion and to continue to monitor antimicrobial resistance.Item Atividade de compostos naturais e sintéticos na presença de espécies de Candida(Universidade Federal de Goiás, 2017-02-09) Andrade, Fernanda Almeida; Costa, Carolina Rodrigues; http://lattes.cnpq.br/2503738140054176; Costa, Carolina Rodrigues; Silva, Maria do Rosário Rodrigues; Oliveira, Valéria de; Passos, Xisto Sena; Costa, Maysa Paula daInfections caused by yeasts of the genus Candida are called candidiasis and are considered opportunistic diseases with characteristics ranging from superficial to disseminate. Candida species may develop defense mechanisms to the major antifungal agents available for clinical use against infections, making it difficult and limiting treatment. Natural products can be a source of bioactive compounds against infectious diseases, such as the Curcuma longa Linn or saffron plant as it is popularly known that it has the compound curcumin. This vegetable is used in large scale by the population as food condiment and as natural medicine against inflammatory diseases and infectious diseases. Laboratory-synthesized compounds have been to interest in medical biotechnology, among which the hydrazone derivatives are associated with antimicrobial including antifungal, antioxidant, antiparasitic, anticonvulsive, analgesic, anti-inflammatory and antitumor activity. The objectives were to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of curcumin and n-acylhydrazone compounds, and also to verify the time-kill curve of Candida, to quantify ergosterol and to evaluate hemolysis of red blood cells after action of both compounds. Twenty isolates Candida spp. in the broth microdilution test for evaluation of the MIC and MFC of the compounds. The time-kill curve and ergosterol assay were checked after treatment of C. albicans ATCC 28367 with the compounds. To evaluate the cytotoxicity of these compounds sheep red blood cells were used. The compound curcumin showed MIC values between 16 and 256 μg/mL and MFC between 128 and values greater than 512 μg/mL. For the n-acylhydrazone derivative the MIC values ranged from 2 to 128 μg/mL and the MFC variation ranged from 256 to values greater than 512 μg/mL. The compounds showed fungistatic action on the growth kinetics of Candida over 48 hours. The determination of ergosterol after contact with the agents in MIC of 64 μg/mL of curcumin and 16 μg/mL of nacylhydrazone was reduced by 29.56 and 53.57%, respectively. The compounds showed hemolytic activity at concentrations of 512 μg/mL. The results observed in this work show that the curcumin and the n- acylhydrazone derivative have promising antifungal properties.Item Avaliação do comportamento metabólico de cisticercos de Taenia crassiceps cultivados in vitro e expostos a condições estressantes(Universidade Federal de Goiás, 2013-09-06) Andrade, Lilian Cristina Morais de; Vinaud, Marina Clare; http://lattes.cnpq.br/1921551651088660; Vinaud, Marina Clare; Costa, Tatiane Luiza; Castro, Ana Maria deTaenia crassiceps cysticerci possess antigenic similarities to T. solium cysticerci and because of this and added to the relative easiness in its maintainance in laboratory they are used as an experimental model to T. solium studies. The objective of this study was to analyze the in vitro influence of glucose and insulin on the energetic and respiratory metabolism of T. crassiceps cysticerci exposed to low dosages of anti-helminthic drugs, albendazol and praziquantel, and also to different concentrations of glucose and insulin. Glucose is the main energy source to these parasites in their larval and adult stages and glycogen is their main energetic reserve. Glycogen may be converted into glucose in case of low glucose uptake and is stored in the cysticerci tegument. The cysticerci were collected from the peritoneal cavity of BALB/c female mice experimentally infected and maintained in the animal facilities of IPTSP/UFG. After 24hours of culture in RPMI culture medium supplemented and with glucose, glargine insulin (LANTUS®, Sanofi Aventis), albendazol and praziquantel, the cysticerci were removed and the medium was frozen in liquid nitrogen as to allow the metabolix stasis. Afterwards this culture medium was analyzes through HPLC as to permit the quantification of the following organic acids related to the carbohydrates metabolism: lactate and pyruvate, intermediary metabolism: citrate, α-ketoglutarate, succinate, fumarate, malate and oxaloacetate, fatty acids metabolism: β-hydroxibutyrate and propionate. It was possible to detect propionate and β-hydroxibutyrate secreted by T. crassiceps cysticerci which indicates the fatty acids oxidation as an alternative energy xv source used by the initial stage cysticerci which are in rapid growth. Also the detection of organic acids from the citric acid cycle indicates the in vitro aerobiosis performed by the initial stage cysticerci.Item Identificação e caracterização das proteínas da parede celular em Paracoccidioides sp.(Universidade Federal de Goiás, 2014-02-24) Araújo, Danielle Silva; Parente, Ana Flávia Alves; http://lattes.cnpq.br/8717846564577423; Soares, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637; Soares, Célia Maria de Almeida; Bailão, Alexandre Melo; Baeza, Lilian CristianeParacoccidioidomycosis (PCM) is the important systemic mycosis in Latin America, with high incidence in Brazil, Argentina, Colombia and Venezuela. The disease has been attributed to the thermodimorphic fungus Paracoccidioides sp.. During the infective process, we can highlight the role of the cell wall, which is a dynamic structure, vital for growth, survival and morphogenesis of the fungus. In addition, this structure is constantly changing in response to environmental signals and different stages of the cycle of the fungus. The interest in the fungal cell wall occurs primarily by lack of this structure in mammalian cells; for this reason cell wall components are promising targets for antifungal drug design. In order to describe the profile of cell wall proteins (CWPs) of Paracoccidioides sp., it was used a proteomic approach coupling nanoscale liquid chromatography to multiplexed mass spectrometry (nanoUPLC-MSE) to identify the CWPs isolated from Paracoccidioides sp. yeast cells and mycelia. Among the identified proteins, it was found a transglycosylase orthologue to the Crh1p, which is a GPI anchored protein, known to be involved in attaching chitin to β-glucan. Adhesins previously described in Paracoccidioides sp. such as enolase, glyceraldehyde-3-phosphate, alcohol dehydrogenase, fructose-1,6-biphosphate aldose and some chaperones were also identified. Moreover, we identified formamidase that was previously described as localized in the fungus cell wall and may be involved in nitrogen metabolism besides contributing with antigenic properties.Item Leishmaniose Tegumentar Americana na região Centro-Oeste: avaliação de dados clínicos, epidemiológicos, laboratoriais e moleculares(Universidade Federal de Goiás, 2014-04-30) Balian, Rosana Pereira Morais; Dorta, Miriam Leandro; http://lattes.cnpq.br/3933395097851681; Dorta, Miriam Leandro; Moraes, Sandra do Lago; Loyola, Patrícia Resende Alo NagibProtozoa of the genus Leishmania, which affects the skin and/or mucous membranes, cause American Cutaneous Leishmaniasis (ACL). It is an endemic zoonosis whose numbers of cases in the Brazil Midwest region are growing. In the Goiás state, 2798 ACL cases were been reported between 2007 and 2013. In the present study, our proposal was to investigate the epidemiological and molecular characteristics of ACL patients attended at the ambulatory of the Hospital Anuar Auad in the period 2000 at 2006 and identify the species of Leishmania sp. The study included 152 patients with ACL, 124 were from of the state of Goiás and 28 at Mato Grosso, aged between 6-79 years, and center and thirteen these individuals were male. For diagnosis, clinical, epidemiological and laboratory data were collected, such as direct examination (ED), histopathology (AH), Montenegro skin test (MST), indirect immunofluorescence (IIF). ELISA using crude extract of L. (Viannia) braziliensis was performing. Characterization of Leishmania species was carried out by polymerase chain reaction (PCR). The positivity of ED, AH, MST, IIF and ELISA was 70.6%, 80.9%, 68.9%, 44.2% and 73.0%, respectively. Specific IgG to L. (V.) braziliensis were detecting in 84.7% of patients with mucosal leishmaniasis (ML), significantly higher than those found in patients with cutaneous leishmaniasis (CL), which was 69.0% (p < 0.05). Detection of IgG before and after treatment was performed using ELISA and was observed a statistically significant difference only in samples obtained from patients with CL after 6 and 18 months of treatment. The data serological monitoring of patients with ACL before and after treatment indicates that the total IgG levels tend to decrease after 6 months of treatment. PCR was performed on 69 samples obtained from scraping the edge of lesions of patients with ATL, these, 62 (89.8%) were positive, 53 patients with LC and 9 patients with ML. The samples were characterized by PCR as L. (V.) braziliensis (93.5%) and L. (L.) amazonensis (6.5%). It was found that 25.8% of cases of leishmaniasis in the state of Goiás occurred in Campestre, Goiânia, Aparecida de Goiânia and Niquelândia. By analyzing the frequency of cases of ATL over the months during the years analyzed, it was observed that there was a distribution of ACL in all seasons from 2000 to 2006, with a larger number of cases in winter. PCR represents important and powerful tool in the diagnosis and species identification in ACL in endemic areas. ACL controlling in endemic areas is difficult and requires an accurate idea of its epidemiology.Item Nanopartículas poliméricas e interações com macrófagos(Universidade Federal de Goiás, 2016-09-16) Bandeira, Anielle Carvalho; Amaral, André Corrêa; http://lattes.cnpq.br/8801299423520104; Amaral, André Corrêa; Loyola, Patrícia Resende Alo Nagib; Celes, Mara Rúbia NunesNanotechnology involves the creation and use of materials, devices and systems through control of matter on the manometer scale. It has positive impact on medicine involving the treatment and diagnosis. Drug delivery systems prepared with nanostructures have the ability to overcome biological barriers and optimizing drug release. However, one of the main challenges in the use of these systems is their internalization by macrophages. This study aims to prepare and characterize polymer nanoparticles formed by biodegradable polymers and investigate its impact on the function of macrophages in vitro by the observation of cell viability, phagocytic activity and cytokine production. Also, was investigate the cytokine profile in serum and splenocytes culture supernatant from Balb/C mice injected with chitosan nanoparticles. The chitosan and chitosan-containing magnetic nanoparticles were prepared by ionic gelation crosslinking, resulting in a morphology "nearly spherical" and positive zeta potential. The PLGA prepared by emulsification and solvent evaporation method presented spherical morphology and negative zeta potential. All preparations had a diameter smaller than 300 nanometers and polydispersity index lower than 0.5. Nanoparticles of chitosan and PLGA affected the viability of macrophages in vitro only at the highest concentration tested (4 mg/mL). The chitosan nanoparticles containing magnetic nanoparticles were internalized by macrophages after 4h and 24h of incubation. Different nanoparticles triggered high production of TNF- and low IL12p40 production and IL-10. Serum and splenocytes culture supernatant was observed in both the production of IFN- as IL-4. However, IL-4 production was higher than that of IFN-.Item Astrovírus humanos clássicos em amostras fecais de crianças atendidas em um hospital de Goiânia, Goiás(Universidade Federal de Goiás, 2019-04-23) Barbosa, Gabriela Rodrigues; Souza, Menira Borges de Lima Dias e; http://lattes.cnpq.br/0054562567103606; Souza, Menira Borges de Lima Dias e; Fiaccadori, Fabíola Souza; Baur, CarmenClassical human astroviruses are classified into the family Astroviridae, genus Mamastrovirus, and are further classified into eight serotypes / genotypes (HAstV 1-8). These viruses are considered important agents of non-bacterial acute gastroenteritis (GEA), and can infect individuals of all age groups, being predominant in children up to five years of age. The objectives of this study were to evaluate the occurrence and estimate the viral load of classical human astrovirus, and to perform molecular characterization of positive samples from faecal samples obtained from children up to six years of age with symptoms of acute gastroenteritis (vomiting and / or diarrhea, with or without abdominal pain, with or without fever). For this real-time reverse transcription polymerase chain reaction (RT-qPCR)with standard recombinant plasmid curve and specific probes and primers specific for RLA2 genomic region were used, followed by genomic sequencing and phylogenetic analysis of the positive samplesFecal samples were obtained from 250 children attending a referral hospital in Goiânia from May 2014 to April 2015. A global positivity index of 3.2% (8/250) was observed for HAstV in the feces, and of these positive children, 50% (4/8) presented symptoms of GEA. The viral load ranged from 2.8x105 CG / mL to 1.6x1011 CG / mL, with an average of 2.39x1010. The molecular characterization of the positive samples was performed, of which four were sequenced, two as HAstV-1, 1-a lineage and two as HAstV-4, 4-c lineage. The highest detection occurred in the month of May (5/8), and no defined circulation pattern was observed in relation to dry and rainy seasons. The data obtained in this study contributes to a better understanding of the molecular epidemiology of these agents in the children of the region.Item Citomegalovírus em pacientes submetidos a transplante de células progenitoras hematopoiéticas(Universidade Federal de Goiás, 2016-04-15) Borges, Francielly Pinheiro da Silva; Souza, Menira Borges de Lima Dias e; http://lattes.cnpq.br/0054562567103606; Souza, Menira Borges de Lima Dias e; Carneiro, Megmar Aparecida dos Santos; Saddi, Vera AparecidaThe Human Cytomegalovirus (HCMV) is an important cause of morbi-mortality in recipients of allogeneic hematopoietic stem cell transplantation (AloHSCT). However, there is not a consensus on which protocol to use for monitoring the infection by HCMV and, data on the frequency and clinical manifestations of the infection in this group population are quite variable among the distinct transplant centers in the world. Thus, the main objective of the present study was to proceed the monitoring of active HCMV infection in patients undergoing AloHSCT by three different methodologies: antigenemia (AGM), nested-PCR (nPCR) and real-time PCR (qPCR) and determine viral load, correlating active infection with the clinical manifestations and prognosis of patients. For this, 21 patients undergoing AloHSCT were monitored (from pre-transplant period -5 days prior to transplantation- until one year after transplantation). For HCMV detection three methodologies were used: AGM, nPCR and qPCR, and for molecular detection a comparison was made between detection of HCMV in DNA extracted from pellet (buffy coat) and serum, in a paired manner. The results showed that the active HCMV infection was detected by at least one of three methodologies in 95.2% (20/21) of patients and 45% (9/20) of these were positive in pre-transplantation period, having been observed good agreement between the results of AGM and qPCR (kappa = 0.65). Of the 20 patients positive for active HCMV infection, 85% (17/20) were positive for the three methods and only 15% (3/20) were positive for AGM and qPCR, and negative by nPCR. Regarding the type of clinical sample, molecular techniques showed higher sensitivity to the pellet over the serum. The main alteration of patients was pancytopenia and the main complication was graft-versus-host disease. Six patients died during the study period, however, it was not possible to confirm if HCMV active infection was directly associated with the cause of death. The obtained data reveal a high positivity index and the occurrence of HCMV syndrome in patients submitted to aloHSCT. We hope that the results may assist in the therapeutical measures, as well as in the methodology of choice and the type of clinical sample for detection of active HCMV infection, in order to contribute for the inclusion of HCMV monitoring is included in the routine testing of patients.Item Atividade larvicida do extrato bruto de Swinglea glutinosa evidenciada pelas alterações morfohistológicas em larvas de Aedes aegypti (Diptera, Culicidae)(Universidade Federal de Goiás, 2015-06-29) Cabral, Sávio da Silva; Silva, Ionizete Garcia da; http://lattes.cnpq.br/5021551669347602; Silva, Ionizete Garcia da; Arruda, Walquíria; Silva, Heloisa Helena Garcia da; Silva, Ionizete Garcia daAedes aegypti is the major vector of dengue, urban yellow fever and emerging arboviruses in Brazil, as chikungunya fever. This mosquito is adapted to the urban environment, and is an important factor for the maintenance of its kind, the long period of quiescence of eggs, which was an extraordinary physiological adaptation to different climatic and dietary factors. Approximately 2.5 billion people live in endemic, exposed to dengue transmission countries. The best solution would be to control multivalent vaccines to existing serotypes. Tetravalent vaccines are being tested in several countries, including Brazil. However, recently it described a fifth serotype, and this can probably extend the course of research. So how vaccines are not ready for use, dengue control has been done by the intervention of actions against the vector, Ae. aegypti. Among the actions, the use of synthetic chemical insecticides has been the main, it emerged mosquito susceptibility of modification in many Brazilian cities and later confirmation of resistance status. This paper proposes the search for alternatives, through prospecting of natural compounds insecticides for the control of Ae. aegypti. Among the actions, the use of synthetic chemical insecticides has been the main. However, with the continuous use of the same product in some brazilians municipalities seemed evidence of mosquito susceptibility modification and subsequently confirm the resistance status. Among, measures control alternatives Ae. aegypti is prospecting for natural compounds insecticides. This work proposes the study of crude ethanol extract (c.e.e.) the result of Swinglea glutinous as larvicide and it showed CL50 and CL90 of 74.4 ppm to 125.2 ppm for third instar larvae of Ae. aegypti, besides the toxicity effect on these intestinal cells larvae investigated by morphohistological study. The morphological changes that are intensified with increasing time of exposure to the larvae of c.e.e. were visualized particularly in the posterior region of the midgut, included folding of the peritrophic matrix, increasing subperitrophic space, cytoplasmic vacuolization, reduction of the microvilli cells. What characterized and checked the larvicidal effect of c.e.e. demonstrating its potential use for controlling Ae. aegypti.Item Identificação de astrovírus não clássicos na região centro-oeste(Universidade Federal de Goiás, 2019-06-26) Caetano, Amanda Campos; Souza, Menira Borges de Lima Dias e; http://lattes.cnpq.br/0054562567103606; Souza, Menira Borges de Lima Dias e; Matos, Márcia Alves Dias de; Silva, Hugo Delleon daThe classical human astrovirus (HAstV-1 to HAstV-8) are viruses commonly associated with gastroenteritis in children up to five years of age. Recently new astrovirus species have been recognized and designated as novel astrovirus, Melbourne (HAstV-MLB) and Virginia (HAstV- VA), which have also been found in association with gastroenteritis and other more severe clinical conditions such as meningitis and encephalitis. However, there are still few studies on the occurrence of these pathogens in Brazil, and so far without reports in the Midwest region. Thus, for the first time, novel astrovirus was investigated, in this region, in stool samples obtained between May 2014 and April 2015 from children (≤6 years) with or without symptoms of acute gastroenteritis. The children were attended at the Materno Infantil Hospital in the city of Goiânia, Goiás. For the MLB research the reverse transcription polymerase chain reaction (RT-PCR) was used, followed by Semi-Nested PCR targeting region 3 'end of the ORF1b of the viral genome. It was also carried out genomic sequencing and phylogenetic analysis for molecular characterization. From 250 samples tested, one was positive (0.4%) for novel astrovirus and characterized as HAstV-MLB1. The sample was obtained from a 12- month-old child with diarrhea and respiratory symptoms (cough), in addition to fever. This is the first study to detect non-classical astrovirus in faecal samples obtained in the Midwest region. We hope that the data obtained may contribute to a better understanding of the molecular epidemiology and aspects of the pathogenesis of these agents in the pediatric population.Item Investigação molecular de vírus em crianças com infecção do trato respiratório e crianças asmáticas em Goiânia-Goiás(Universidade Federal de Goiás, 2015-04-23) Castro, Ítalo de Araújo; Fiaccadori, Fabíola Souza; http://lattes.cnpq.br/0993842409303174; Fiaccadori, Fabíola Souza; Souza, Menina Borges de Lima Dias e; Costa, Paulo Sérgio Sucasas daAcute respiratory infection (ARI) is a major cause of morbity and mortality worldwide, particularly among children, and most of these infections are caused by viruses. Respiratory viral infections can cause symptoms ranging cough, coryza, sneezing, fever and airflow obstruction. Furthermore, the infection poses as an important trigger of asthma exacerbation, frequent clinical condition in children, and its prevalence has been rising in the last years. There are few epidemiologic studies analyzing the relationship between ARIs and asthma in Brazil. Based on this background, the aim of the study was investigate the occurrence of viral respiratory infections in pediatric patients with and without asthma in Goiânia – Goiás. Between august, 2012 and august, 2013 225 nasal aspirates and/or nasal swab samples were obtained from children with four to 14 years old. The samples were screened by Multiplex Nested-PCR for detection of 16 common respiratory viruses. From 225 samples, 42 had at least one virus detected. Samples from four different patients had more than one virus detected. The viral detection rate in ARI patients (25%), exacerbated asthma (16.3%) and stable asthma (14.8%) showed no significant difference. The most frequent viruses detected were Rhinovirus (28.6%), FLUA (11.9%), Adenovirus (11.9%), HBoV (11.9%) and RSVA (9.5%). The monthly detection rate was higher during the rainy season, period marked by great rainfall and high relative air humidity. Among the positive samples, RSV were detected during the great rainfall months and high air humidity, while the FLU and HBoV were detected during the winter months, period with low air humidity in the Mid-West region. The seasonal profile from the other viruses was unclear. The obtained results reinforces the importance of the viral pathogens in the pediatric population. Although the viral detection rate was not statistically significant, the presence of these pathogens in children is an important matter for consideration, especially to delineate control and prevention measures concerning ARIs and its impact on preexistent asthma. Hence, this study is the first of the kind in the region, and the data provided tried to fill the knowledge gaps about seasonality and circulation of these pathogens.Item Staphylococcus aureus em tonsilas de pacientes com faringotonsilite aguda recorrente: prevalência, perfil de suscetibilidade e caracterização genotípica(Universidade Federal de Goiás, 2013-10-17) Cavalcanti, Veraluce Paolini; Braga, Carla Afonso da Silva Bitencourt; http://lattes.cnpq.br/7673897995590123; André, Maria Cláudia Dantas Porfirio Borges; http://lattes.cnpq.br/1475834090578722; André, Maria Cláudia Dantas Porfirio Borges; Cardoso, Juliana Lamaro; Cardoso, Alessandra MarquesThe bacterial pharyngotonsillitis are infections of the upper airways that occurs predominantly in children and adolescents. Due to the composition of the oral microbiota is difficult to clarify the role of each organism in the etiology of the disease. The presence of bacteria that produce beta-lactamase interferes with the effectiveness of beta-lactam antibiotics, the most commonly drug used in treatment of these infections, promoting the recurrence of the disease. S. aureus is one of the most common pathogen in the etiology of tonsillitis and its relevance is due to the ability of antimicrobial resistance and persistence in the tissues of the tonsils. Tonsillectomy is indicated in cases of recurrent tonsillitis after several failures in the antibiotic terapy. The aim of this study was to determine the prevalence of S. aureus in the tonsils of patients undergoing tonsillectomy, in a teaching hospital in Goiânia, the antimicrobial susceptibility profile and the genetic characterization of isolates. Tonsils obtained from 123 patients were processed, the microorganisms identified and submitted to antibiogram by conventional techniques. The isolates that presented cefoxitin resistance were submitted to tests to determine the minimum inhibitory concentration - MIC for oxacillin and to detect the presence of the mecA gene. All isolates were subjected to PCR for detection of Panton-Valentine leukocidin gene and to PFGE to determine the genetic similarity among them. It was identified 60 S. aureus isolates from 49 patients (39.8%). There were no significant difference in prevalence by sex and, the average age of male patients was lower (8.2 years) (p<0.001) than the female patients (15.3 years). Nine out 49 patients(18.4%) presented two or more different S. aureus isolates. The isolates presented resistance of 85.0%, 10.0%, 15.0%, 3.3%, 10.0%, 3.3%, 18.3% and 8.3% to penicillin, amoxicillin + ácido clavulânico, cefoxitin, ceftriaxone, erythromycin, trimethoprim/sulfamethoxazole, ciprofloxacin and tetracycline, respectively. All isolates were sensitive to linezolid and rifampin. Six erythromycin-resistant isolates (10.0%) showed inducible resistance (MLSbi) to clindamycin and quinupristin/dalfopristin. Eight isolates (13.3%) were resistant to three or more classes of antimicrobials. Despite the resistance to cefoxitin be considered a marker of the presence of the mecA gene only in two resistant isolates it has been found, xv suggesting that the cefoxitin resistance should be mediated by other mechanisms, such as the overproduction of beta-lactamases. None of these isolates showed resistance to more than two classes of antimicrobials. Among the sixty S. aureus isolates, only carried the gene encoding the Panton-Valentine leukocidin. This isolate presented resistance to five classes of antimicrobials and phenotype D. PFGE analysis grouped 36 (60,0%) of 60 isolates in 10 clusters (>80% similarity), since no one specific clone was associated with colonization of the tonsils. It was observed different patients carrying S. aureus isolates genetically identical or with a high level of similarity (>80%), suggesting in these cases, a common origin. The high prevalence of S. aureus in tonsils suggests an ability to colonize the surface and/or the persistence in the tissues of the tonsils. The isolation of MDR bacteria can promote cross-resistance to other bacteria commonly associated with recurrent tonsillitis. The results point to change the paradigm of diagnosis and treatment of recurrent tonsillitis in order to enable the correct use of antimicrobials to reduce the recurrence which is the main cause of tonsillectomy.Item Desenvolvimento e caracterização de sistema nanoestruturado contendo miconazol e farnesol para o tratamento de candidíase vulvovaginal(Universidade Federal de Goiás, 2016-04-15) Costa, Adelaide Fernandes; Amaral, André Correa; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4753997T1; Amaral, André Correa; Souza, Lúcia Kioko Hasimoto e; Ribeiro, Evandro LeãoVulvovaginal candidiasis (VVC) is caused mainly by opportunistic fungus Candida albicans and its yeast to hyphae transition is considered the major virulence factor of this pathogen. The increased incidence of VVC has highlighted the importance of developing new therapeutic strategies. The objective of this study was to develop a mucoadhesive nanostructured system comprising miconazole, and farnesol within chitosan for the treatment of VVC. The drug showed the antifungal miconazole expected efficacy with minimal inhibitory concentration (MIC) of 1 μg/mL. The farnesol quorum-sensing molecule was capable of inhibiting hypha-transition yeast at levels greater or equal to 300 μM. When tested together, farnesol has no effect compared to the MIC obtained for miconazole. Nanoparticles of chitosan-containing miconazole and farnesol were prepared by ionic gelation and showed favorable characteristics for use on mucous membranes, such as diameter less than 300 nanometers (nm), polydispersion index (PDI) less than 0.3, positive zeta potential and acid pH. The encapsulation efficiency (EE) of the nanoparticles was on average 81.1% for miconazole, 31.9% farnesol and 32.7% and 70.0% for miconazole and farnesol when co-encapsulated, respectively. The nanoparticles showed instability as the diameter and PDI, but were stable compared to the EE. Regarding toxicity in cultured fibroblasts (Balb/ c 3T3) were considered non-toxic. The nanoparticles showed antifungal activity against C. albicans strain used, with MIC of 2.5 μg/mL and 2 μg/mL for nanoparticles with miconazole and miconazole/ farnesol, respectively. Nanoparticles containing farnesol inhibit yeast-hyphae transition at concentrations greater than or equal to 240 uM. The antifungal activity in vivo was assessed in the murine model for VVC. Although there is no statistically significant difference between treatments in relation to the counting of colony forming units (CFU), the results suggest that chitosan nanoparticles containing miconazole and farnesol were effective for inhibiting fungal proliferation and chitosan nanoparticles containing farnesol were capable of decreasing the pathogenicity of infection demonstrated by the absence of inflammation.Item Avaliação da produção de interleucina 6, fator de necrose tumoral e interleucina 10 em hemoculturas de pacientes com Leishmaniose Cutânea localizada(Universidade Federal de Goiás, 2017-03-10) Costa, Ana Carolina Vieira da; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Cardoso, Ludimila Paula Vaz; Pereira, Ledice Inacia de AraujoAmerican Tegumentary Leishmaniasis (ATL) is caused by protozoa of the genus Leishmania. The parasite is hosted by macrophages in cutaneous or mucosal lesions. Monocytes are precursors of macrophages, but there is little information about monocyte activation during ATL. The present study aimed to evaluate cytokine profile of monocytes from patients with localized cutaneous leishmaniasis (LCL) in whole blood cultures stimulated with toll-like receptor (TLRs) and NOD2 receptor agonists. Peripheral blood from patients with active lesions and no treatment, and from healthy blood donors paired by sex and age, was diluted into culture medium and incubated with lipopolysaccharide (LPS), TLR4 agonist; lipopeptide Pam3Cys (Pam), TLR2 agonist; muramyldipeptide (MDP), NOD2 agonist or Leishmania (V.) braziliensis antigen (Ag). After incubation, supernatants were collected to measure IL-6 and TNF (6 h) and IL-10 (24 h) by ELISA. There was production of IL-6, TNF and IL-10 in cultures from patients and controls after stimulation with all agonists and Ag (p < 0.05, medium vs stimulus, n = 29). No differences were detected between concentrations of TNF and IL-10, but IL-6 was higher produced in non-stimulated cultures from patients than controls (p < 0.05, n = 29) or when the stimulus was MDP (p < 0.05, n = 19). The combination Pam/MDP increased the production of IL-6, TNF and IL-10 in cultures from patients and controls when compared with each stimulus isolated (p < 0.05, n = 13 - 14); similarly, Ag and MDP together induced higher concentrations of IL-6, TNF and IL-10 (IL-6 and IL-10, especially in cultures from patients) in comparison with each stimulus (p < 0.05, n = 11 - 14). There was no correlation between the number, size or time of lesions and levels of IL-6, TNF or IL-10. Patients with more than one cycle of treatment to obtain clinical cure showed higher levels of MDP-induced IL-6 than those cured with one cycle of treatment (p < 0.05, n = 17 cured with one cycle vs 8 cured with more than one cycle). Data suggest that LCL patient monocytes can be activated via TLR2, TLR4 or NOD2 to produce pro-inflammatory (IL-6, TNF) and anti-inflammatory (IL-10) cytokines and IL-6 can be associated with tissue lesion and delay of lesion healing indicating an immunopathogenic role in LCL.Item Pesquisa de bocavírus humano em pacientes submetidos a transplante alogênico de células progenitoras hematopoiéticas(Universidade Federal de Goiás, 2018-08-17) Costa, Brunno Câmara Lopes; Souza, Menira Borges de Lima Dias e; http://lattes.cnpq.br/0054562567103606; Souza, Menira Borges de Lima Dias e; Fiaccadori, Fabíola Souza; Santos, Norma Suely de OliveiraHuman Bocavirus (HBoVs) are classified in the Parvoviridae family and are associated with respiratory and gastrointestinal symptoms. Viral infections are an important cause of morbimortality in immunocompromised patients such as allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients. The aim of the present study was to evaluate the positivity rate and loads of HBoVs in clinical samples (feces and sera) of patients who were subjected to allo-HSCT at a reference center for bone marrow transplantation in Goiânia, Goiás. A total of 105 fecal samples and 145 sera samples were collected from 21 consecutive patients, during October 2012 to October 2014. Samples were screened by qPCR TaqMan assay, with specific probe and primers targeting all HBoVs genotypes (HBoV-1 to -4), and viral loads were determined using serial dilutions of a recombinant plasmid, targeting the NP1 gene. The results showed that 53.4% (11/21) of the patients were male, aged between four and 61 years-old (mean 35 years). The most observed hematologic malignancy was myeloid leukemia (acute or chronic), accounting for 57.1% (12/21) of the cases. The HBoVs were detected in 42.9% (9/21) of the patients and 77.7% (7/9) were positive in both fecal and serum samples. The viral load in fecal samples were higher than in the sera samples and a prolonged fecal shedding were observed, with two patterns: one intermittent and another continuous. Of all HBoV positive patients, six (66.6%) had the first positive sample before the transplantation, and a rise of the viral loads after the allo-HSCT occurred when comparing to the loads before the allo-HSCT. Furthermore, on most cases the highest viral loads were detected during the first 100 days after the allo-HSCT. Considering the symptoms presented by the patients, 66.6% (6/9) had diarrhea at the same period of the viral genome detection in feces, but no statistical significance was observed. Three fecal samples were characterized as being HBoV-1, with more than 99% of nucleotide identity among them. The present data shows a high occurrence and loads of HBoVs in allo-HSCT recipients, with first positivity in fecal samples and later viral detection in sera. These results suggest that fecal samples could be the sample of choice in HBoV monitoring of these patients both before and after the transplant.Item Análise bioquímica da niclosamida no metabolismo de cisticercos de Taenia crassiceps(Universidade Federal de Goiás, 2015-03-09) Costa, Marco Vítor Silva de Melo; Costa, Tatiane Luiza da; http://lattes.cnpq.br/8219041935358486; Vinaud, Marina Clare; http://lattes.cnpq.br/1921551651088660; Vinaud, Marina Clare; Silva, Luciana Damacena; Bezerra, José Clecildo BarretoExperimental studies with Taenia crassiceps have been used to demonstrate its metabolic alterations in biochemical pathways (energetic and respiratory) in response to the presence of drugs. The use of T. crassiceps as an experimental model offers conditions to reproduce the neurocysticercosis infection one of the most severe form of human cysticercosis. This study aimed the analysis of the in vitro influence of niclosamide, an antihelminthic drug, on the metabolism of carbohydrates and fatty acids of T. crassiceps cysticerci (ORF strain). 20 larval stage cysticerci were cultured into 5mL of RPMI (Gibco) supplemented culture media, in 6 well culture plates, exposed or not to niclosamide (1, 2, and 3 uM). The control groups were performed with cysticerci not exposed to the drug, exposed to ethanol in the concentration used to dissolve the drug. The cysticerci were cultured at 37ºC for 24 hours. After this period the cysticerci were removed from the culture medium and both were frozen with liquid nitrogen. Afterward, the samples were processed for the HPLC analysis. Accordingly to the mode of action of this drug, which is to interfere in the electrons chain transport, the succinate concentrations were altered in the secretion/excretion of this parasite. Therefore, it is possible to conclude that in the concentrations used, this drug caused little alteration in the metabolic pathways of the parasite.Item Caracterização filogenética de isolados do vírus dengue em Goiânia, Goiás(Universidade Federal de Goiás, 2015-04-17) Cunha, Marielton dos Passos; Fiaccadori, Fabíola Souza; http://lattes.cnpq.br/0993842409303174; Fiaccadori, Fabíola Souza; Coelho, Alexandre Siqueira Guedes; Cardoso, Divina das Dôres de PaulaDengue viruses (DENV) serotypes 1, 2, 3, and 4 have been causing yearly outbreaks in Brazil. Nevertheless, the population structure of the viruses transmitted in Goiás state is not well understood. In this study, we investigated the phylogenetic pattern of DENV samples identified in Goiânia, Goiás, Brazil during the 2012/2013 epidemic. Therefore, the entire region of the gene of the envelope (E) protein (1485bp) of 16 DENV-1 samples as well as partial region of this gene (363bp) of seven DENV-4 samples were sequenced. Phylogenetic analysis showed that DENV-1 belongs to the genotype V and presents itself divided into two clades, suggesting co-circulation of two distinct lineages in this region. Still, the molecular analyzes indicated a significant change in the amino acid level E348 position. For DENV-4, the sequences were segregate in a monophyletic group and are classified as genotype II American subclade. The molecular and phylogenetic analysis showed that the region suffered multiple introductions by dengue virus. This is the first report of co-circulation of two lineages of DENV-1, and the circulation of DENV-4 in Goiás state.Item Isolamento e purificação parcial de L-asparaginase produzida por actinobactéria em solo do cerrado(Universidade Federal de Goiás, 2015-07-13) Ferreira Neto, Petain Jose; Vieira, José Daniel Gonçalves; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782223Z6; Vieira, José Daniel Gonçalves; Ribeiro, Evandro Leão; Lima, Ana Beatriz MoriThe Cerrado is the second largest biome in Brazil. Such an environment has a different soil several other regions. There are various living beings in this biome, among them, the actinobacteria. They are microorganisms whose cell wall Gram-positive, >50% G+C (guanine + cytosine) in its DNA, produce substances with high economic value. Actinobacteria are major producers of enzymes useful to humans. L-asparaginase enzyme is used in acute limphoblastic leukemias therapy (ALL) also being used in the pre-treatment of foods rich in reducing sugars, inhibiting the formation of acrylamide. The aim of this study was to produce L-asparaginase enzyme isolated from a soil actinobacteria, characterize the enzyme and identify the species of bacteria isolated. A soil sample from Cerrado was collected and submited to a pretreatment. Chemotaxis of Actinoplanetes spores used for the selection of this organism. Nineteen morphspecies were recovered. We used a medium inducing L-asparaginase, and the pink halo around the colony measured. Isolated BC-A.1 produced the largest halo in less time, with an average enzyme content of 4.1. After the completion of the production steps, partial purification and optimization of enzymatic activity, it was realized that the enzyme produced by isolated morphospecies is very efficient at 40 ° C and pH 8.0. Biochemical, physiological and molecular tests were performed, and the observation with optical microscopy and scanning electron microscopy, but it was not possible to identify the species isolated, probably belongs to the genus Streptomyces.Item Ação da curcumina e morina em leveduras do Complexo Cryptococcus neoformans(Universidade Federal de Goiás, 2017-05-12) Freitas, Vivianny Aparecida Queiroz; Fernandes, Orionalda de Fátima Lisboa; http://lattes.cnpq.br/9826992750728262; Fernandes, Orionalda de Fátima Lisboa; Souza, Lucia Kioko Hasimoto e; Melhem, Márcia de Souza CarvalhoCryptococcosis is a fungal disease caused by yeasts belonging to the complex Cryptococus neoformans. The antifungal arsenal available for the treatment of this disease is still restricted and is related to the high toxicity and side effects of some drugs, causing great harm to the patients. In this context, it is necessary to discover new bioactives to quell the infections and reduce the adverse effects. The plants have rich source of secondary bioactive metabolites such as tannins, terpenoids, saponins, alkaloids, flavonoids and other compounds, registered with expressive antimicrobial properties. Therefore, the research of natural compounds and derivatives of natural products has been relevant in recent years, due to their relevance in the discovery of new drugs, in addition to which the association of drugs or compounds with different mechanisms of action has been used as an alternative in conventional therapy. We evaluated the mechanisms of action and antifungal effect of natural compounds, curcumin and morin, on Cryptococcus neoformans and Cryptococcus gattii isolates. The antifungal activity was determined across the minimum inhibitory concentration (MIC) and minimum fungicide (MFC). Polyphenols showed MICs ranging from 2 to 256 μg/mL. The compounds showed a synergistic interaction in 30% (6/20) and 15% (3/20) of the isolates when dealing with morin and curcumin, respectively, associated with fluconazole. In the interaction with amphotericin B morin presented synergism in 70% (14/20) of the isolates. Both compounds did not exhibit antagonism in any of the combinations. Changes in fungal cell morphology were observed, and in contact with red blood cells, presented low toxicity. The mechanisms of action revealed that both polyphenols act on the membrane of the fungal cell, inhibiting the synthesis of ergosterol and causing damage to it. These results demonstrate the potential of the compounds as natural bioactives, with great impact on the discovery of new drugs and their efficacy to be used in the treatment of fungal infections.