Doutorado em Medicina Tropical e Saúde Pública (IPTSP)
URI Permanente para esta coleção
Navegar
Navegando Doutorado em Medicina Tropical e Saúde Pública (IPTSP) por Por Área do CNPQ "CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA"
Agora exibindo 1 - 6 de 6
Resultados por página
Opções de Ordenação
Item Surto de infecção após videoscopias causado por Mycobacterium massiliense em Goiânia-GO : análise molecular e determinação da suscetibilidade aos antimicrobianos(Universidade Federal de Goiás, 2009-12-03) CARDOSO, Alessandra Marques; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; KIPNIS, André; http://lattes.cnpq.br/4434965360286741In recent years the number of infections caused by microbacteria non-tuberculous mycobacteria (NTM) has increased mainly due to opportunistic infections in individuals imunocompormetidos and improvement of farming techniques and identification of MTN. Mycobacterium massilienese is an emerging body associated with wound infections, abscesses and pneumonia. An outbreak of infection after videoscopy occurred between 2005 and 2007 in seven hospitals in Goiânia-GO, in central Brazil. The objective of this study was to identify NTM isolated from patients with infection after arthroscopy and lararoscopia by PCR followed by analysis of fragment length polymorphism restrção (PRA-hsp65), compared by gel electrophoresis pulsed-field gel (PFGE), sequencing of the partial rpoB gene and determination of antimicrobial susceptibility in vitro. NTM were recovered from samples (exudate abscess subcutâneio) of 18 patients involved in the outbreak. In the period leading up to this study there was no reported case of infection after videoscopy caused by MTN in Goiania. The 18 isolates were identified as M, massiliene and genotyped as a single clone, indicating that they had a common origin, suggesting a common source of infection for the patients involved in the outbreak. The epidemic isolates were susceptible to amikacin (MIC90 4 micrograms / ml) and clarithromycin (MIC90 <1 ug / ml), but resistance to ciprofloxacin (MIC90 <128g/ml), tobramycin (MIC90 32 micrograms / ml) and intermediate susceptibility to cefoxitin (MIC90 64 ug / ml). In conclusion this study demonstrated the clonality of strains of M. massiliense involved in infections after procedures videoscopes and that they are susceptible to drugs indicated for the treatmentItem Caracterização bioquímica da endoxilanase recombinante (HXYN2r) do fungo termofílico Humicola grisea var. thermoidea e sua aplicação na sacarificação de resíduos agrícolas(Universidade Federal de Goiás, 2008-06-30) CARVALHO, Wagner Rodrigues de; FARIA, Fabrícia Paula de; http://lattes.cnpq.br/3739169267521003Xylanases have been used in the biobleaching of paper pulps, in bioconversion of plant biomass, for food and feed industries, among others. For successful selection of xylanases suitable for specific industrial applications it is important to characterize enzymes isolated from different sources. The thermophilic fungus Humicola grisea var. thermoidea is described as a good producer of extracellular endoxylanases and the Hxyn2 gene from this fungus was isolated and expressed in yeast Pichia pastoris. The aim of this project was focused on the production, purification and biochemical characterization of the recombinant HXYN2 (HXYN2r) enzyme and application on enzymatic hydrolysis of lignocellulosics substrates. The culture conditions of P. pastoris in flasks, using the 12.3 transformant and BMMY-U medium, were optimized and the best xylanolytic activity was 478.2 U/mL after 96 h, with a protein concentration of 100 mg/L, using 2.34% (w/v) of nitrogen sources (yeast extract plus urea 1.34:1.0% - w/v), 1% (v/v) of methanol, and OD600 of 10. The HXYN2r enzyme was purified by gel filtration chromatography with a yield of 6.4% and showed optimum pH and temperature values of 6.5 and 60ºC, respectively, maintaining 100% of initial activity after 4 h of incubation at pH 5.5, 6.5 and 7.5. The half-life time was 18 min at 60ºC and the enzyme was 100% stable after 3 h of incubation at 50ºC. Km and Vmax values were 7.9 mg/mL e 235.4 μmol/(mL.min), respectively. The HXYN2r enzyme were used on the enzymatic hydrolysis of sugarcane bagasse (SCB), corn cob (CC) and foliar sample of sugarcane (FSC) alone or with the enzymes of xylanolytic and cellulolytic system produced by H. grisea fungus. For enzymatic hydrolysis experiments, the substrates were milled and pretreated with 0.25% (w/v) of H2SO4 by 30 min. On the enzymatic hydrolysis the best conversion yield of hemicellulosic fractions were obtained using PpHXYN2r supplemented with EHg: 42.8% to CC, 9.6% to SCB and, a mean of 20% to foliar samples.Item Identificação e caracterização de moléculas envolvidas na interação de Paracoccidioides brasiliensis com o hospedeiro(Universidade Federal de Goiás, 2009-03-31) DANTAS, Sabrina Fonseca Ingênito Moreira; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM), a systemic mycosis presenting clinical manifestations ranging from mild to severe forms. A P. brasiliensis cDNA expression library was produced and screened with pooled sera from PCM patients adsorbed against antigens derived from in vitro-grown P. brasiliensis yeast cells. Sequencing DNA inserts from clones reactive with PCM patients sera indicated 35 open reading frames presenting homology to genes involved in metabolic pathways, transport, among other predicted functions. The complete cDNAs encoding aromatic L-amino acid decarboxylase (Pbddc), lumazine synthase (Pbls) and a homologue of the high affinity copper transporter (Pbctr3) were obtained. Recombinant proteins PbDDC and PbLS were obtained; a peptide was synthesized for PbCTR3. The proteins and the synthetic peptide were recognized by sera of patients with confirmed PCM and not by sera of healthy patients. Using the vivo-induced antigen technology (IVIAT) we identified immunogenic proteins expressed at high levels during infection. Quantitative real - time RT-PCR demonstrated high transcript levels of Pbddc, Pbls and Pbctr3 in yeast cells infecting macrophages. Transcripts in yeast cells derived from spleen and liver of infected mice were also measured by qRT-PCR. Our results suggest a putative role for the immunogenic proteins in the infectious process of P. brasiliensis.Item Suscetibilidade in vitro e alterações morfologicas em células melanizadas e não melanizadas na presença de voriconazol, anfotericina B e extrato bruto da folha de P. pseudocaryophyllus (Gomes) L. R. Landrum(Universidade Federal de Goiás, 2010-10-25) FERNANDES, Orionalda Fatima Lisboa; LINO JÚNIOR, Ruy de Souza; http://lattes.cnpq.br/0372118837748010; SILVA, Maria do Rosário Rodrigues; http://lattes.cnpq.br/7119226630434725Item Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas(Universidade Federal de Goiás, 2010-03-11) NOGUEIRA, Sarah Veloso; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), a human systemic mycosis, prevalent in Latin America. Extracellular matrix (ECM) is a complex net where collagens, laminin and fibronectin can be found and, when exposed, is the first site for the fungus adhesion. Our aim was to study genes involved in the adhesion process using Representational Difference Analysis (RDA). RDA is a PCR-coupled subtractive method that allows the isolation of genes differentially expressed in two different cDNA populations. Hence, cDNAs were synthesized from RNAs extracted from P. brasiliensis yeast cells adhered to collagen and fibronectin to identify overexpressed genes. Genes involved in a wide range of cellular process were found and PbCtr3 (cooper transporter) and enolase (PbEno) were chosen to further studies. A synthetic peptide (PbCTR3) and the recombinant enolase (rPbEno) were utilized together with the anti-rPbEno polyclonal antibody in functional analysis with ECM components and plasminogen. The studies suggest that P. brasiliensis enolase, in the surface, is able to generate plasmin from plasminogen by plasminogen activator. Therefore, it was also demonstrated that this protein is secreted and able to promote fungus adhesion and invasion to cells. These findings clearly establish the role of enolase in the patogenicity of P. brasiliensis.Item Tipagem molecular de Streptococcus pneumoniae isolados da nasofaringe de crianças no contexto da vacinação pneumocócica(Universidade Federal de Goiás, 2010-02-18) ROCHA, Cristyane Gonçalves Benicio Bastos; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899; PIMENTA, Fabiana Cristina; http://lattes.cnpq.br/2230554075502158Objectives (i) Present review article focusing on pneumococcal vaccines and carriage; (ii) to validate sequential multiplex PCR for identifying pneumococcal capsular serotypes from children attending day-care centers; (iii) determine the multilocus sequence typing; (iv) to identify the capsular types of multiple colonies of S. pneumoniae isolates from a single sample of nasopharyngeal secretions of children attending day-care centers in Goiânia. Materials and Methods S. pneumoniae was obtained from health children less than 5 years old attending 62 day care centers of Goiânia. The laboratory procedures were performed according to WHO recommendations. Were selected 217 isolates (penicillin resistant and sensitive) for capsular typing by multiplex PCR technique. MLST was performed for 55 isolates representing the serotypes detected and the different susceptibility patterns for penicillin. Quellung reaction was used for typing isolates serotypes 6A, 6B, 18C and the isolates not typed by multiplex PCR. For 28 presumptive pneumococcal positive NP swabs, 3 colonies were picked to acess possible serotype diversity. Eighty four pneumococci were identified by conventionally procedure and multiplex PCR was performed. Results Serotypes were deduced for 177/217 (81.6%) of the pneumococcal. The most frequent serogroups/serotypes were 14, 6, 23F, 19F and 18. Multiple serotypes were detected in 13 specimens. Were found 19 MLST types and two new ST. Forty (18.4%) were not serotyped by the multiplex PCR and quellung reaction. The analysis of three colonies from the same NP permitted the detection of differente serotypes in 7/28 (25%) NP samples. Conclusion (i) The multiplex PCR is simple and cost-effective method for detecting multiple serotypes in nasopharyngeal isolates; (ii) and thus might be useful for the monitoring of pneumococcal colonization over time; (iii) the use of multiplex PCR can further broaden our understanding of the dynamics of pneumococcal carriage, including multiple serotypes, the effect of vaccination on carriage, and transmission, as well as surveillance of IPD and co-colonization.