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Enfermidades infecciosas em comunidade indígena Terena de Mato Grosso do Sul
(Universidade Federal de Goiás, 2008-08-02) AGUIAR, José Ivan Albuquerque; NETTO, Joaquim Caetano de Almeida; http://lattes.cnpq.br/3444498706763045
The health conditions of Brazilian indigenous population are few recognized with limited information available. A survey was carried out among indians Terena, people that inhabit on the municipalities of Sidrolândia and Dois Irmãos do Buriti, Estado de Mato Grosso do Sul, Brazil. Was studied the prevalence of infection by intestinal parasites, infection markers for viral Hepatitis A, B and C, and for antibody against the viral diseases; Poliomyelitis 1, 2 and 3, Measles, Yellow Fever and Hepatitis B. The results were stratified by age and revealed that the parasitic infection affects 73.5% of the population studied, with high prevalence of the Blastocystis hominis. In the population above 10 years, more than 90% showed reactivity to the anti-HAV, absence of infection markers for Hepatitis B and C, respectively HBsAg and anti-HCV, and a rate of 16.7% (95% CI 12.6-21.3) for the anti-HBs. The prevalence of neutralizing antibodies against the measles virus and yellow fever virus was 96.7% (95% CI 93.9-98.3) and 91.4% (95% CI 88.0-94.7) respectively. The polio results showed that 62.2% (95% CI 56.5-67.6) 71.7% (95% CI 66.2-76.6) and 63.5% (95% CI 56.5-69.6) had antibodies against the types 1, 2 and 3, respectively, showing vulnerability to B Hepatitis, Yellow Fever and Poliomyelitis.
Epidemiologia molecular do HIV-1, resistência aos antirretrovirais em gestantes e transmissão vertical no estado de Goiás
(Universidade Federal de Goiás, 2010-10-31) ALCÂNTARA, Keila Correia de; STEFANI, Mariane Martins de Araújo; http://lattes.cnpq.br/5581414958714905
Introduction: The spread of the aids epidemic among young women and HIV-1 mother-tochild transmission (MTCT) represent important public health issues. In this context, prenatal care represents a unique opportunity for the early diagnosis of young women and for the implementation of full preventive strategies to HIV-1MTCT. Objectives: To study immunological, virological, clinical and epidemiological characteristics and to identify factors associated with HIV-1MTCT among HIV-1 pregnant women/infants recruited in Goias State. Material and methods: Cohort 1: 41 pregnant HIV/AIDS: infant pairs (April/2000-August/2001) were recruited and prospectively followed up at two regional reference centers-Mother-Infant Hospital (HMI/SUS) and Hospital Dr. Anuar Auad (HAA/HDT/SUS). Cohort 2: 172 HIV/AIDS pregnant women and 149 exposed children were recruited at the Institute of diagnosis and prevention (IDP/APAE) and prospectively followed up at HAA/HDT/SUS. The following tests were performed: maternal viral load, CD4+ T cell counts, HIV-1env/gag subtypes by heteroduplex mobility assay/HMA (cohort 1) and pol (protease and reverse transcriptase-PR/RT) sequencing for resistance profile, subtypes identification and phylogeography analysis for subtype C (cohort 2). Infants born to HIV-1/aids mothers were evaluated by plasma viral RNA and CD4+ T cell counts. Seroreversion of exposed- uninfected children was followed by sequential ELISA tests for IgG anti HIV-1. Results: Patients from cohorts 1 and 2 presented similar social-demographic and clinical profiles. The median age was 26 years; 15-41 years), lower educational level predominated and most were diagnosed during pregnancy (90%). Over 80% received ARV prophylaxis. One case of MTCT was observed in cohort 1 which was associated with short prophylaxis and long labor period. Exposed-uninfected infants born to symptomatic mothers seroreverted earlier. Cohort 2 included 80% of all HIV-1 infected pregnant women from Goias state in that period. The early prophylaxis and undetectable viral load predominated among previously diagnosed patients (p<0.05). One ARV naive patient presented transmitted drug resistance; 10 ARV experienced patients presented secondary drug resistance: 6 under MTCT prophylaxis, 4 under HAART. MTCT was observed in 3/149 (2.01%) cases and late diagnosis, vaginal delivery, brastfeeding and lack of oral ZDV were observed. Among MTCT cases resistance mutations were not detected. HMA env/gag (cohort 1) and pol sequencing (cohort 2) results showed mostly subtype B followed by subtypes F1, C and recombinants, mainly BF1. HIV-1 subtype C was identified only among pregnant women from cohort 2 which together with recombinants BC represented around 20% of the isolates. Subtype C and BC recombinants were isolated in interior municipalities of Goias state located close to the main highways that connect south/southern to north (BR153), northeast (BR020) and South/west (BR369/BR070). Phylogenetic/ phylogeographic analysis showed a subtype C clado, clusters (aLTR ≥ 0.85) with sequences from Southern states and from Sao Paulo and evidences of multiple introductions. Conclusion: Our results indicate the importance of prenatal care for the early diagnosis/prevention of HIV-1 vertical transmission. However late diagnosis and missed opportunities to fully prevent transmission were associated with vertical transmission. Multiple introductions and the dissemination of HIV-1 subtype C by heterosexual contact in interior cities highlight the importance of monitoring the genetic diversity and the impact of subtype C dissemination in the interior of Brazil. Note: superscript + is where it appears and the program does not copy.
Caracterização de microrganismos isolados em manipuladores e dietas enterais de dois hospitais públicos de Goiânia
(Universidade Federal de Goiás, 2010-03-19) BORGES, Liana Jayme; ANDRÉ, Maria Cláudia Dantas Porfirio Borges; http://lattes.cnpq.br/1475834090578722; SERAFINI, álvaro Bisol; http://lattes.cnpq.br/9849440763539845
Enteral feeding means the nutrition for special purposes, with controlled intake of nutrients. The advantages of its use often become secondary to complications arising from its contamination, which may be associated with infectious complications. The microbial contamination of enteral feeding may occur during all steps being the handling, particularly critical. Considering the importance of enteral feeding as a therapeutic tool in hospitals and the need to guarantee the microbiological quality of the products offered to critical patients, the present work aimed to evaluate the hygienic and sanitary quality of diets and their ingredients and to identify and characterize phenotypic and genotypically, using the antibiogram and pulsed-field gel electrophoresis, strains of Escherichia coli and Staphylococcus aureus obtained from handlers hands and noses, water, module and enteral nutrition from two public hospital in Goiânia, Brazil in order to investigate the probable source of microbological contamination. A total of 80 samples were collected from enteral nutrition and 140 from hands and noses of handlers involved in the diets manufacturing in hospital 1 (H1), between october/2007 and november/2008 and 80 samples from enteral nutrition and 80 from hands and noses of handlers in hospital 2 (H2), between october/2008 and november/2008. From both hospitals were collected 40 samples from water and module. The samples were submitted to microbiological analysis to verify the presence and numbers of pathogenic and indicator microorganisms. E. coli and S. aureus strains were submitted to antibiogram and PFGE. According to antibiogram, all S.aureus isolates (15) from H1 were susceptible to oxacillin, vancomycin, ciprofloxacin and gentamicin. Resistence profile was observed in 10 (66.7%) isolates for penicillin, four (26.7%) isolates for tetracycline and nine (60.0%) isolates for erythromycin, allowing to classify the strains in six different phenotypes (A-F), but it was not efficient for the determination of the bacterial source for the diets. In the H1, all (08) E. coli strains were susceptible to trimethoprim, ciprofloxacin, cephalothin, gentamicin, ceftazidime and tetracycline. Resistence was observed in six (75.0%) isolates for ampicilin. In H2, all strains isolated (12) were susceptible to trimethoprim, ciprofloxacin, gentamicin and ceftazidime and resistence was observed in 11 isolates (91.7%) for cephalothin and 12 (100.0%) for tetracycline and ampicillin, grouping them into five different phenotypes (A-D). Microorganisms showed the same phenotypic profile from handlers and diet samples (phenotypes A and C), suggesting that in these cases, the source of microorganisms for the final product was the food handler. The genotypic typing of S. aureus strains by PFGE generated seven different DNA banding profiles and the E. coli genotyping generated five profiles. Based on the results, two E. coli strains isolated from diets were identical to one strain isolated from food handler from H2 and two of S. aureus isolated from diets were identical to one strain isolated from food handler from H1. This study shows that the enteral feedings showed unsatisfactory sanitary-hygienic conditions in both hospitals and the hand contact is probably one of the sources of greatest significance for enteral diets contamination in the hospital environment.
Validação da gastrostomia endoscópica percutânea realizada pela técnica de punção com o uso de uma nova variante técnica de gastropexia
(Universidade Federal de Goiás, 2010-12-16) CAMPOLI, Paulo Moacir de Oliveira; TURCHI, Marília Dalva; http://lattes.cnpq.br/3769826743537934
Percutaneous endoscopic gastrostomy (PEG) currently represents the main alternative to ensure nutritional supply in patients with prolonged or permanent inability to swallow, and yet has a functional gastrointestinal tract. PEG performed with the Pull Technique is widely used because it is easy to perform and very safe, although it is associated with high infection rates. The Introducer Technique appears to be associated with a lower infection risk, although it requires fixation of the gastric wall to the abdominal wall (gastropexy), which makes the procedure difficult to perform. This study sought to describe and validate PEG performed with the Introducer Technique with the use of a new technical gastropexy variant, besides demonstrating its benefits in relation to risk of peristomal infection. A descriptive study of the safety and feasibility of a new technical gastropexy variant with a long curved needle was performed. We then compared the two gastropexy techniques in a before-and-after design. A randomized clinical trial comparing gastrostomy tubes constructed of different materials (latex vs. silicone) was also conducted. Finally, we performed a meta-analysis evaluating peristomal infection risk between the Introducer Technique and Pull Technique. The results of these four studies are presented in four separate papers. The first study showed that the new technical gastropexy variant that uses a long curved needle is safe and feasible. The second study showed that it is associated with a lower risk of infection compared with the gastropexy technique used previously. The third study found that the silicone tubes have greater durability than latex tubes. The final meta-analysis showed that PEG performed with the Pull Technique is associated with a greater risk of infection than the Introducer Technique.
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/4434965360286741
In 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 treatment
Epidemiologia molecular e riscos associados ao portador nasal de Staphylococcus aureus isolados de crianças de creches de Goiânia
(Universidade Federal de Goiás, 2009-04-15) CARDOSO, Juliana Lamaro; KIPNIS, André; http://lattes.cnpq.br/4434965360286741; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899
Objectives: (i) to assess the prevalence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) nasal carriage in children attending day-care centers (DCC) in the municipality of Goiânia; (ii) to determine the potential risk factors related to S. aureus carriage and MRSA; (iii) to characterize MRSA isolates circulating in DCCs using molecular typing methods. Methods: Between August and December 2005, nasal swabs were collected from children who attended 62 DCCs. Clinical and socio-demographic information associated with the acquisition of S. aureus and MRSA were obtained through questionnaires applied to parents or guardians. The swabs were processed following the standard methods for identification and isolation of S. aureus. Amplification femB gene by polymerase chain reaction (PCR) was used to confirm the specie. The presence of mecA gene was detected by PCR and the positive isolates were identified as MRSA. Susceptibility to MRSA was determined by disk diffusion method. MRSA molecular typing was performed by PFGE, MLST, spa typing and SCCmec multiplex PCR. Results: 371 (31.1%) out of the 1.192 collected swabs were positive for S. aureus and 14 (1.2%) were identified as MRSA. The factors independently associated with risks for nasal colonization by S. aureus were children higher than two years of age (OR = 1.83, 95% CI 1.27-2.65) and previous DCC attendance (OR = 1.48; 95% CI 1.01-2.16). Mother s high degree of education was a protective factor for S. aureus carriage (OR = 0.43, 95% CI 0.23-0.80). A multidrug resistant dominant MRSA lineage was identified comprising 8 out of the 14 MRSA isolates. This cluster was characterized as SCCmec type IIIA, ST239 and spa type t037 sharing 82.7% genetic similarity with the Brazilian clone. One MRSA strain was classified as SCCmec type V and ST1120. This strain showed features of CA-MRSA although it has been recovered from a healthy child who presented risk factors for HA-MRSA acquisition. The remaining MRSA strains showed a diverse genetic background. Conclusions: Children attending DCCs are often colonized with S. aureus and although the prevalence of MRSA was low, they can represent potential vectors of spread of resistant pathogens to the community. The detection of a MRSA lineage circulating within DCCs suggests a two-way flow spread of MRSA between hospitals and community.
Estratégias na identificação e caracterização de potenciais antifúngicos e seus alvos em Paracoccidioides brasiliensis
(Universidade Federal de Goiás, 2010-09-10) CARVALHO, Patrícia Fernanda Zambuzzi; PEREIRA, Maristela; http://lattes.cnpq.br/1345781867765758
The termodimorphic fungus Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis (PCM), a systemic human mycosis geographically distributed in Latin America, being the eighth most common cause of death among chronic infections. PCM is acquired by inhalation of fungal propagules, which reach the lungs and is disseminate through the bloodstream and/or lymph to all parts of the body. The treatment of PCM is long, starting with a dosage of aggressive antifungal agents, extending for months or years. Resistance to antimicrobial drugs may limit the ability of effective treatment of patients, interfering with therapeutic efficacy. Thus, it is necessary to discover and develop new antifungal agents. Plants compounds have a great structural diversity, many of which are models for the synthesis of a vast number of drugs. The action of the oenothein B compound, purified from leaves of Eugenia uniflora, a plant from the Brazilian Savannah, was evaluated on growth, viability and expression of P. brasiliensis genes. The compound interfered with cell morphology and inhibited the transcripts of β-1- 3-glucan synthase. The synergistic effect between oenothein B and drugs used to treat PCM (amphotericin B, itraconazole, Sulfamethoxazole and Trimethoprim-Sulfamethoxazole Combination) was evaluated in this study by the method of sensitivity on plates. The highest inhibition of the fungal growth was observed in association of oenothein B with Trimethoprim-Sulfamethoxazole Combination, followed by amphotericin B, itraconazole and sulfamethoxazole, respectively. Representational Difference Analysis (RDA) was also performed to elucidate the mechanism of action of oenothein B in P. brasiliensis. We identified 463 ESTs up regulated and 121 ESTs down regulated after 90 min of incubation of P. brasiliensis yeast cells with the compound. After 180 min incubation 301 ESTs up regulated and 143 down regulated were identified. The ESTs were classified according to their functional categories using the program Blast2GO. The analysis indicated the presence of transcripts with functions related to cell wall and membrane, transcription factors and hypothetical proteins. In this study, we evaluated also the characteristics of the malate synthase (Pbmls) cDNA, regulation of Pbmls gene expression, and enzymatic activity of the MLS protein of P. brasiliensis (PbMLS), isolate Pb01. The cDNA contains 1617 bp, which encodes a protein of 539 amino acids. The protein has the signature of the MLSs, residues essential for catalytic activity and addressing signal for peroxisomes, PTS1. The high level of Pbmls transcript observed in the presence of 2C sources suggests that in P. brasiliensis, the primary regulation of carbon flux into glyoxylate cycle was at the level of the Pbmls transcript. Transcript analysis, protein levels and enzymatic activity in the presence of different carbon and nitrogen sources suggest that PbMLS acts in both pathways: in glyoxylate cycle, when 2C sources are used, and in alantoin degradation pathway, when proline is used as nitrogen source, or when oxalurate is used to induces genes from pathway.
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/3739169267521003
Xylanases 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.
Estudo do metabolismo energético de Leishmania spp utilizando cromatografia líquida de alta eficiência e ressonância magnética nuclear
(Universidade Federal de Goiás, 2011-09-16) COSTA, Tatiane Luiza da; VINAUD, Marina Clare; http://lattes.cnpq.br/1921551651088660; BEZERRA, José Clecildo Barreto; http://lattes.cnpq.br/9491755585617846
Leishmaniasis is a disease of the endothelial reticulum system and is caused by protozoans form the Leishmania genus. The promastigote forms of Leishmania sp degradate carbohydrates through the glycolityc pathway and its first reaction occurs inside the glicosome. As they use as energy source glucose, aminoacids and lipids from the culture media the excreted/secreted (E/S) products may be useful as indicative of the energetic and metabolis profile of the parasite and of its biological disturbances. The biochemical analysis of Leishmania metabolism is important to upgrade the parasitological studies with techniques which are adequate to the evaluation of the biochemical profile of the host-parasite relationship towards a better understanding of how these organisms are able to survive in such diverse environments such as the digestory tract of the insect or inside vertebrate hosts tissues. The aim of this study was to perform a study of the energetic metabolism of species and isolates of Leishmania spp. Therefore the in vitro analysis of E/S metabolic products from the parasite within the culture media during logarithmic and stationary growth phases was performed through chromatographic (HPLC) methods. The spectrophometry was used to glucose measurement and the nuclear magnetic resonance (NMR) was implanted aiming the study of the parasite‟s metabolism. Through HPLC it was possible to verify a significant difference in the E/S of organic acids from the carbohydrates metabolism (pyruvate and lactate), intermediary metabolism (citrate, α-ketoglutarate, succinate, fumarate, malate and oxaloacetate) and from the fatty acids oxidation (β-hydroxibutyrate) between differents species and isolates of Leishmania spp. Through the enzymatic spectrophotometric analysis we verified the different concentrations of glucose. The NMR through 13C and 1H detected the following metabolites in pellet and supernatant: alanine, lactate, acetate, oxalate, pyruvate, succinate, glicerol, CO2, ureia and aminoacids (glutamine, valine and arginine). Therefore, through the several methods it was possible to observe that there is difference of E/S metabolites between the isolates and strains of Leishmania spp that utilize several biochemical pathways to energy production and the identification of these pathways may be used to identify each isolate and also establish metabolic differences in the host-parasite relationship.
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/8539946335852637
Paracoccidioides 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.
Epidemiologia molecular dos vírus dengue em Goiânia-GO, 1994 - 2006: vigilância laboratorial e caracterização dos sorotipos circulares
(Universidade Federal de Goiás, 2008-08-06) FÉRES, Valéria Christina de Rezende; MARTELLI, Celina Maria Turchi; http://lattes.cnpq.br/5867052489026059
Nowadays, dengue constitute the major public health problem, because is relevant cause of illness and death between thousands people that resident in the tropical and subtropical regions in world. The dengue virus is classified as four serotypes (DENV-1, 2, 3 and 4) according to antigenic differences and characterized intra-typical groups called genotypes. The laboratorial surveillance enables the diagnostic confirmation of dengue infection and monitoring serotypes circulating through the routine diagnostic techniques. Recently, the use molecular techniques has contributed to characterize and monitoring of the genotypes potentially virulent during epidemic and knowledge of biology of dengue virus. This thesis was organized in an introduction section, that include a literature review on dengue, and two manuscripts that describing the research conducted with focus on laboratory diagnostic and molecular epidemiology. The first manuscript entitled Laboratorial Surveillance of Dengue Virus in Central-Brazil, 1994-2003, was published at Journal of Clinical Virology, 2006 37 (3): 179-83. In this study we present the results of the virological surveillance for dengue cases conducted in the city of Goiânia (~1,200,000 population) from 1994 to 2003. Suspected cases were from the main public infectious disease reference hospital and outpatient clinics covering the metropolitan area. Serological and virus isolation tests were conduced at the regional reference laboratory. Our objective was to report dengue circulating serotypes from 1994 to 2003 and the role of distinct serotypes on dengue clinical outcomes in Central Brazil and to characterize serotypes and genotypes by reverse transcriptase PCR (RT-PCR) and by restricted site-specific PCR (RSS-PCR) patterns in selected samples. Laboratory surveillance identified mainly DEN-1 serotype from 1994 to 2002 shifting to a high circulation of DEN-3 in 2003. The adults (87,4%) were the most affected group and dengue fever accounted for the majority of the cases. Diagnosis of dengue was confirmed in ~50% of the suspected and enhanced by RT-PCR. RSS-PCR patterns for DEN-1 and DEN-3 corresponded to the circulating subtypes in the country. The infection DENV-3 did not suggest a major role of infecting DEN-3 in increasing disease severity during its first-year spread in Central Brazil. The second manuscript, to be submitted for publication is entitled: Epidemiologia Molecular do Vírus Dengue tipo 3 em Goiânia GO, 2005-2006. The objective of this manuscript was to characterize the DENV-3 genotype circulating isolated from well-characterized clinical and laboratory samples in Goiânia-GO/Brasil. Seven samples had sequences of the prM/M/E region obtained and comparative analysis was performed with the reference strains. The results showed the homology of the genomics sequences with genotype III strains. The nucleotide identity of the all the samples varied from 97.0% to 99.6% and the amino acid sequences from 97.5% to 99.5%. The analysis of the nucleotide sequence revealed silent mutation and 14 amino acid changes in the protein deduced from gene prM/M/E. In conclusion, the study confirms that the strains of DENV-3 in Central Brazil relate to genotype III. The genomic changes along Domain III of the protein E were observed, which could affect the pathogenicity, but were not consistent between samples of DCC and DHF. Samples of patients with dengue fever had mutations related to viral attenuation. More investigation is necessary to evidence of genomic changes found in relationship with clinical forms.
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/7119226630434725
Detecção de vírus gastroentéricos em mulheres em Goiânia-GO
(Universidade Federal de Goiás, 2009-04-30) FERREIRA, Rui Gilberto; CARDOSO, Divina das Dôres de Paula; http://lattes.cnpq.br/9770835116155857
The gastroenteric viruses are important etiological agents of gastroenteritis in individuals of all ages. It is believed that individuals with deficits in the immune system (humoral and/or cellular), among them pregnant women and human immune deficiency virus (HIV)-seropositive women, are more susceptible to these viral infections. The rotaviruses, adenoviruses, astroviruses, and caliciviruses constitute are among the main causes of acute gastroenteritis in the world, and are accounted for high morbi-mortality rates, especially among children under five years of age. It is believed that, by the age of three, approximately 90% of all children in developing countries have antibodies to one or more of these agents. Neonatal infection does not exclude the possibility of re-infection, with different viral serotypes, however it protects the individual against severe disease. This study aimed at the detection of rotavirus, adenovirus, astrovirus e calicivirus in women seeing at the Hospital das Clínicas da Universidade Federal de Goiás (HC-UFG) and at the investigation of as association between the positivity to this viruses and the low immune status, characteristic in pregnant women and/or HIV-seropositive women. This was a prospective follow-up study of women seeing at the Gynecology and obstetrics (OB-GYN) sector of the HC-UFG aiming at the detection of gastroenteric viruses (rotavirus, adenovirus, astrovirus e calicivirus). For this, fecal samples were collected from 84 women, in the period from July-2006 to June-2007. For rotavirus detection, fecal samples were screened by polyacrilamide gel electrophoresis (PAGE) and by an immunoenzimatic assay (IEA). The calicivirus and astrovirus were detected by polymerase chain reaction post-reverse transcription (RT-PCR), and adenovirus detection was performed by an EIE. The astrovirus genotyping was conducted by Nested-PCR. Three-hundred and fourteen fecal samples were collected from a total of 84 women. From those 84 women, 29 were HIV-seropositive, 55 HIVseronegative, 45 were pregnant at the time, and 39 were not pregnant. The patients were aged between 16 and 67 years-old, and 47% of them had up to 30 years of age. From the total 84 patients, 19 (22.6%) were positive for calicivirus and/or astrovirus in at least one of the collected samples, as follows: calicivirus (14/19) and astrovírus (6/19), with the highest positivity rates being detected in the months of July and August (astrovirus) and September and October (calicivirus). None of the collected samples were positive for rotavirus or adenovirus. The association index between gastroenteric virus positivity and pregnancy, in the presence or not of HIV-seropositivity, was of 68.4% (13/19); however, there was no significant difference between the group of women that were not pregnant and the ones that were HIV-seronegative. The gastroenteric viruses were detected in a significant parcel (22.6%) of this population of adult women, and were represented by the caliciviruses (16.7%) and astroviruses (7.1%). Under the conditions of the present study, no association was found between pregnancy and/or positivity for HIV, as reducing factors for the immunological capacity of the women, and the detection of gastroenteric viruses; in conclusion, the pregnancy and/or HIV-seropositivity did not increase the chances of these women to be infected by these gastroenteric viruses.
FATORES DE RISCO E EPIDEMIOLOGIA MOLECULAR DE Streptococcus pneumoniae NÃO SUSCETÍVEIS À PENICILINA ISOLADOS DE NASOFARINGE DE CRIANÇAS QUE FREQUENTAM CRECHES EM GOIÂNIA-GO, BRASIL
(Universidade Federal de Goiás, 2009-02-17) FRANCO, Cáritas Marquez; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899
Objectives: (i) to identify risk factors for S. pneumoniae penicillin nonsusceptible isolates (PNSp) in children attending day-care centers (DCCs) in Goiânia, Brazil and to assess the genetic patterns of pneumococcal isolates; (ii) to estimate the coverage for carriage serotypes for the 7-valente (PCV7) pneumococcal conjugate vaccine, and for the investigational 10 (PCV10) and 13-valent (PCV13) vaccines; (iii) to assess the genetic relatedeness between isolates expressing capsular type 14 and those non(sero)- typeable isolates (NTPn); (iv) to investigate if carriage isolates match genetically to any international pneumococcal clone (PMEN network). Methods: A cross-sectional survey of carriage PNSp was conducted among 1.192 children, 2 months to 5 years of age, attending 62 DCCs in Central Brazil. Capsular typing was performed in PNSp isolates (CLSI, 2007) and in a sample of isolates susceptible to penicillin (PSSp) matched to PNSp and DCCs whenever possible. Serotyping was performed by Quellung reactions and confirmed by multibead assay. NTPn isolates and serotype 14 were tested by PCR for capsule genes. Odds ratio for PNSp carriage and respective 95% confidence interval (95%CI) were assessed by logistic regression. Pulsed field gel electrophoresis (PFGE) was applied to assess the genetic similarity between PNSp serotype 14 and NTPn isolates. PCR was performed for the presence of pneumococcal capsule gene locus. For comparison purpose we also evaluated the genetic profile of PNSp serotype 14 invasive strains derived from the current pneumococcal invasive disease surveillance for the same pediatric population. Isolates were epidemiologically related if they shared ≥80% similarity on the dendrogram (Dice coefficient). A cluster was defined as three or more related isolates. Results: A total of 686 pneumococci were isolated for a colonization rate of 57.6% and 178 (25.8%) were PNSp. Among the PNSp isolates the usual common types were found: 14 (53%), 23F (10.2%), 6B (6%), 19F (4.8%) and 19A (4.2%). PSSp isolates displayed 30 different serotypes although serotype 14 was the most common. Overall a high prevalence of NTPn (11.1%) was observed with 62.9% PNSp. Serotypes coverage xvi for the PCV7, PCV10 and PCV13 vaccines were 55.2%, 55.9% and 65.1%, respectively. Being less than 24 months of age (OR=1.79; p=0.006), hospitalization in the previous three months (OR=2.19; p=0.025), and recurrent acute otitis media (OR=2.89; p=0.013) were independently associated with PNSp in a multivariate model. Among the 123 PNSp submitted to PFGE (106/carriage and 17/ invasive isolates) a major group of 34 serotype 14 strains (8 invasive and 26 carriage) was identified and found to be genetically related to the global pneumococcal clone Spain 9V-3 (82.7% similarity). All NTPn presented capsule gene locus and 10 (45.4%) of them presented capsule gene locus to type 14. Conclusions: (i) DCC attendees with history of recurrent AOM could significantly contribute to the spread of nasopharyngeal PNSp strains into the community; (ii) epidemiologic and molecular evidences support the findings that pneumococcal nonypeable carriage isolates are genetically similar to carriage and invasive isolates expressing capsular type 14; (iii) carriage and invasive isolates circulating in Goiânia belong to a serotype 14 variant of the Spain 9V -3 clone and play a critical role in the spread of PNSp strains to the entire pediatric community of Goiânia
Investigação epidemiológica e molecular da infecção pelo vírus da hepatite C em usuários de drogas ilícitas no Brasil Central
(Universidade Federal de Goiás, 2009-04-02) LOPES, Carmen Luci Rodrigues; MARTINS, Regina Maria Bringel; http://lattes.cnpq.br/2582896795892370
Hepatitis C virus (HCV) infection is an important problem of public health. Drug users (DU) constitute a group of frequent exposure to HCV and little is known about this infection in DU in Brazil. This study aimed to investigate the seroepidemiological and molecular profile of HCV infection among drug users in Central Brazil. A total of 691 DU, being 102 injection drug users (IDU) and 589 noninjecting drug users (NIDU), were interviewed and blood samples collected in 26 treatment drug centers in Campo Grande-MS and Goiânia-GO. Blood samples (sera) were tested for antibodies to HCV (anti-HCV). Anti-HCV-positive samples were submitted to HCV RNA detection by polymerase chain reaction (PCR) with primers complementary to the 5 NC and NS5B regions of viral genome and genotyped by LiPA and direct nucleotide sequencing followed by phylogenetic analysis, respectively. The anti-HCV prevalence was 6.9% (95% CI: 5.2-9.2). The study population reported a low known regarding HCV transmission ways, such as parenteral (20.8-30.5%), sexual (31,7%) and vertical (20%) of HCV. Multivariate analysis of risk factors revealed that age > 40 years, route (injecting) and duration of drug use and blood transfusion were associated with HCV infection. HCV RNA was detected in 85.4% of the anti-HCV-positive samples. Thirty-three samples were of genotype 1 by the LiPA, subtypes 1a (63.4%) and 1b (17.1%), and eight samples (19.5%) were of genotype 3, subtype 3a. The phylogenetic analysis of the NS5B region showed that 17 (68%), 5 (20%) and 3 (12%) samples were of subtypes 1a, 3a and 1b, respectively. This study shows a high HCV infection prevalence and the predominance of subtype 1a among drug users in Central Brazil. Also, the injecting drug use was the risk factor most strongly associated with this infection. In addition, the study population reported a low known regarding HCV transmission ways. Thus, preventive measures are needed to control this infection in illicit drug users
Estudo epidemiológico e molecular da infecção pelo vírus da hepatite B em Afro-descendentes de comunidade isolada no Estado de Goiás (Kalungas)
(Universidade Federal de Goiás, 2007-12-18) MATOS, Márcia Alves Dias de; MARTINS, Regina Maria Bringel; http://lattes.cnpq.br/2582896795892370
Hepatitis B virus (HBV) infection occurs throughout the world. In Africa, this infection is highly endemic, with the majority of individuals becoming infected during childhood. Although Brazil has been globally considered a country of HBV intermediate endemicity, variable rates have been found in all five Brazilian regions and even inside the same region. This study aimed to investigate the epidemiological and molecular profile of the HBV infection among the Kalunga population in Goiás, Central Brazil, which is considered the largest Afro-Brazilian isolated community. A total of 878 individuals were interviewed about sociodemographic characteristics, risk factors and HBV vaccination. Blood samples were collected from all participants and serum samples were screened by enzyme-linked immunosorbent assay for the presence of HBsAg, anti-HBc and anti-HBs serological markers. HBsAg-positive samples were submitted to HBeAg and anti-HBe detection. HBsAg and anti-HBc positive samples were tested for HBV DNA detection by polymerase chain reaction and genotyping by subsequent restriction fragment length polymorphism (RFLP) analysis and nucleotide sequencing of preS/S region. The overall prevalence of HBV infection was 35.4% (95% CI: 32.3-38.7). HBsAg carrier rate was 1.8% (95% CI: 1.1- 3.0). Multivariate analysis of risk factors showed that increased age, male gender, illiteracy and history of multiple sexual partners were associated with this infection. Isolated anti-HBs was found in 301 (34.3%) individuals who were immune for hepatitis B. HBV DNA was detected in 75% (12/16) of the HBsAg positive samples, in 100% (2/2) of the HBeAg and in 83.3% (10/12) of the anti-HBe positive samples. An occult HBV infection rate of 1.7% (5/295) was found among anti-HBc positive individuals. All genotyped isolates belonged to genotype A by RFLP analysis. Nucleotide sequencing of preS/S region confirmed the circulation of genotype A (subgenotype Aa) in this community. The epidemiological findings indicate that preventive measures, such as additional health education and HBV vaccination programs, are needed to control HBV infection in this population. In addition, the molecular results suggest the introduction of genotype A, subgenotype Aa in Brazil from Africa during the slave trade.
Pneumonia e pobreza, uma abordagem espacial: Dois anos de vigilância populacional do estudo LEAP-Brazil
(Universidade Federal de Goiás, 2012-12-06) MELO, Lícia Kamila Assis; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899
Mortalidade por causas violentas em jovens de Goiânia
(Universidade Federal de Goiás, 2010-05-19) MINAMISAVA, Ruth; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899
Background: Deaths due to homicides and traffic accidents among youth are a public health issue worldwide. Studies of the complex network of cause and effect on this topic point to both poverty and health inequalities. Different investigational approaches to intentional and unintentional deaths combined with socioeconomic variables can help create a better understanding of the association between violence and socioeconomic conditions. This study analyzed the spatial distribution and potential clusters of risk for intentional and unintentional deaths among youths aged 15-24 years in Goiânia, a newly urbanized city in central Brazil. Methods: Death data and residential addresses were extracted from the national Mortality Information System and validated by household visits. To detect all potential cases, we prospectively investigated every death classified as a transport accident, assault, legal intervention,intentional self-harm, unknown underlying cause, and undetermined intent according to the ICD-10. The Geographical Information System was used to plot residential addresses, and cases were interactively geocoded to the residential address level using a digital map of the municipality. Spatial scan statistic was applied (Poisson model) to identify clusters of census tracts with high mortality due to intentional injuries and traffic accidents. The socioeconomic variables obtained using census data were compared between the most likely cluster and other areas of the municipality. Results: The most violent deaths among young people were due to intentional injuries. Between August 2005 and August 2006, 145 addresses for cases of intentional injuries and traffic accidents were located and geocoded. No significant clusters for deaths due to traffic accidents were found within the municipality. One significant cluster (RR = 4.65; p = 0.029) composed of 14 cases of intentional deaths, mostly homicides, was detected in an emergent, populated, and very poor area on the outskirts of the town. This cluster had a significantly higher proportion of people with the lowest educational status, lowest income, and poor housing conditions in comparison to the remainder of the municipality. Conclusion: Our findings highlight the link between social inequalities and intentional deaths, clearly showing the need for urgent social interventions to reduce violence and premature mortality.
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/8539946335852637
Paracoccidioides 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.
Contribuição ao estudo da doença de Chagas
(Universidade Federal de Goiás, 2011-07-18) OSTERMAYER, Alejandro Luquetti; NETTO, Joaquim Caetano de Almeida; http://lattes.cnpq.br/3444498706763045
The main papers published on the last five years on the area of human Chagas disease, mainly on parasitological, serological and therapeutic aspects, reflecting areas of recent involvment of the author, were selected. As a baseline, briefly comments on publications on the last 40 years were included. Parasitemia profile measured by hemoculture during the chronic infection was the subject of the first two. The first (published in 2006) included six hemocultures from each of 27 patients, the last three after specific treatment. Results were compared with those of 13 non treated, infected patients. The supressive effect of benznidazole was demonstrated in 89% of the patients and treatment failure was registered in three cases (11%) during the two year follow-up. In the second paper (2011) a single hemoculture was performed in 152 infected women, 101 pregnant. Parasitemic pregnant women doubled the number of non-pregnant, mainly during the first months of pregnancy. A new ELISA test was developed (2010) by employing two recombinant proteins and two synthetic peptides. Sensitivity was 99.3% on 165 positive sera, and specificity of 100% (216 negatives). A multicentric study was done (2009) with participation of laboratories of North America (Mexico), Central America (Honduras) and South America (Brazil) with 98 serum samples from patients of Mexico employing reagents made from Tc1 and Tc2 strains. Results showed good agreement among laboratories demonstrating the feasibility of using reagents prepared from both types of T. cruzi. In another multicentric study (Bolivia, Argentina and Brazil) xenodiagnosis performed in 17 patients after a 60 days course of allopurinol treatment, remained positive, showing the lack of effect of this drug in the chronic phase. A national serological survey in children born after insecticide spraying (below five years old) with filter paper, involved 104,954 samples, tested with ELISA and indirect immuofluorescence. Only 11 samples (0.01%) were identified as by vector transmission (negative mothers) mainly from the northeast of the country, areas without Triatoma infestans. These findings (2011) confirm the effectivity of house spraying for the control of the disease. As a sub-product of this investigation, an unusual number of cases of congenital transmission (n = 12) was found only in the State of Rio Grande do Sul, where TcV-TcVI circulates. When compared with other states (n=8) the proportion was 10 times higher. This is the first report on geographical differences related to congenital transmission in Brazil. We considered that all these findings contributed significantly to a better knowledge on different aspects of Chagas disease.

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