Revealing a novel Otubain-Like Enzyme from Leishmania infantum with deubiquitinating activity toward K48-linked substrate

Deubiquitinating enzymes (DUBs) play an important role in regulating a variety of eukaryotic processes. In this context, exploring the role of deubiquitination in Leishmania infantum could be a promising alternative to search new therapeutic targets for leishmaniasis. Here we present the first characterization of a DUB from L. infantum, otubain (OtuLi), and its localization within parasite. The recombinant OtuLi (rOtuLi) showed improved activity on lysine 48 (K48)-linked over K63-linked tetra-ubiquitin (Ub) and site-directed mutations on amino acids close to the catalytic site (F82) or involved in Ub interaction (L265 and F182) caused structural changes as shown by molecular dynamics, resulting in a reduction or loss of enzyme activity, respectively. Furthermore, rOtuLi stimulates lipid droplet biogenesis (an inflammatory marker) and induces IL-6 and TNF-a secretion in peritoneal macrophages, both proinflammatory cytokines. Our findings suggest that OtuLi is a cytoplasmic enzyme with K48-linked substrate specificity that could play a part in proinflammatory response in stimulated murine macrophages

Recomendações para o uso de testes de DNA-HPV no rastreamento do câncer do colo útero no Brasil

Evidence-based clinical guidelines ensure best practice protocols are available in health care. There is a widespread use of human papillomavirus deoxyribonucleic acid (HPVDNA) tests in Brazil, regardless of the lack of official guidelines. On behalf of the Brazilian Association for the Lower Genital Tract Pathology and Colposcopy (ABPTGIC, in the Portuguese acronym), a team of reviewers searched for published evidence and developed a set of recommendations for the use of HPV-DNA tests in cervical cancer screening in Brazil. The product of this process was debated and consensus was sought by the participants. One concern of the authors was the inclusion of these tests in the assessment of women with cytologic atypia and women treated for cervical intraepithelial neoplasia (CIN). Testing for HPV is recommended in an organized screening scenario to identify women with precursor lesions or asymptomatic cervical cancer older than 30 years of age, and it can be performed every 5 years. It also has value after the cytology showing atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesions (LSILs) as a triage test for colposcopy, in the investigation of other cytological alterations when no abnormal findings are observed at colposcopy, seeking to exclude disease, or, further, after treatment of high-grade cervical intraepithelial neoplasia, to rule out residual disease.O uso de diretrizes clínicas baseadas em evidências visa assegurar as melhores práticas na área de cuidado à saúde. O uso de testes de ácido desoxirribonucleico de papilomavírus humano (DNA-HPV) vem crescendo e se disseminando sem que existam recomendações de uso no cenário brasileiro.Emnomeda Associação Brasileira de Patologia doTrato Genital Inferior e Colposcopia (ABPTGIC), grupos de revisores pesquisaram evidências e formularamrecomendações para o uso dos testes de DNA-HPV no rastreamento do câncer do colo do útero, no seguimento de mulheres com atipias citológicas, e após tratamento de neoplasia intraepitelial cervical (NIC). O produto desse processo foi debatido e foi buscado consenso entre participantes. Os testes de DNA-HPV são recomendados num cenário de rastreamento organizado para identificação de mulheres portadoras de lesões precursoras ou câncer assintomático com mais de 30 anos e podem ser realizados a cada 5 anos. Também têm valor após a citologia mostrando células escamosas atípicas de significado indeterminado (ASC-US) ou lesão intraepitelial escamosa de baixo grau (LSIL) como teste de triagempara colposcopia, na investigação de outras alterações citológicas quando não são observados achados anormais à colposcopia, buscando excluir doença, ou, ainda, no seguimento após tratamento das neoplasias intraepiteliais de alto grau, para exclusão de doença residual

Resistência Antimicrobiana de Shigella spp. isoladas no Estado do Pará

Introduction: Shigella spp. are gram-negative, nonsporulating, rod-shaped bacteria that belong to the family Enterobacteriaceae and are responsible for shigellosis or bacillary dysentery, an important cause of worldwide morbidity and mortality. Methods: We studied the antibiotic resistance profiles of 122 Shigella spp. strains (81 S. flexneri, 41 S. sonnei, 1 S. boydii) isolated from patients (female and male from 0 to 80 years of age) presenting diarrhea in different districts of the State of Pará, in the North of Brazil. The antibiotic resistance of the strains, isolated from human fecal samples, was determined by the diffusion disk method and by using the VITEK-2 system. Results: The highest resistance rate found was the resistance rate to tetracycline (93.8%), followed by the resistance rate to chloramphenicol (63.9%) and to trimethoprim/sulfamethoxazole (63.1%). Resistance to at least three drugs was more common among S. flexneri than S. sonnei (39.5% vs. 10%). Six (4.9%) strains were susceptible to all the antibiotics tested. All strains were susceptible to cefotaxime, ceftazidime, ciprofloxacin, nalidixic acid and nitrofurantoin. Conclusions: High rates of multidrug resistance in Shigella spp. are a serious public health concern in Brazil. It is extremely important to continuously monitor the antimicrobial resistances of Shigella spp. for effective therapy and control measures against shigellosisIntrodução: Shigella spp. são bactérias gram-negativas, não esporuladas, em forma de bastonete, pertencentes a família Enterobacteriaceae responsáveis pela shigelose ou disenteria bacilar, uma importante causa de mortalidade e morbidade mundial. Métodos: Foi estudado o perfil de resistência a antimicrobianos de 122 amostras de Shigella spp. (81 S. flexneri, 41 S. sonnei, 1 S. boydii) isoladas de pacientes (sexo feminino e masculino com faixa etária de 0 a 80 anos) com distúrbios gastrointestinais em diferentes municípios no Estado do Pará, Brasil. A resistência antimicrobiana das amostras isoladas de coprocultura, foi determinada pelo método de difusão em disco e pelo sistema Vitek II. Resultados: A maior resistência foi observada em relação à tetraciclina (93,8%), seguida de cloranfenicol (63,9%), e trimetoprimsulfametoxazol (63,1%). Multirresistência a pelo menos três antimicrobianos foi mais comum em S. flexneri comparada a S. sonnei (39,5% vs. 10%). Seis (4,9%) amostras foram sensíveis a todos antimicrobianos testados. Todas as amostras apresentaram sensibilidade acefotaxima, ceftazidima, ciprofloxacina, ácido nalidixico e nitrofurantoína. Conclusões: As altas taxas de multirresistência de Shigella spp. são um sério problema de saúde pública no Brasil. Sendo assim, torna-se extremamente importante um monitoramento contínuo da resistência antimicrobiana de Shigella spp. para uma terapia efetiva e medidas de controle contra shigelose

Antimicrobial Resistance of Shigella spp. isolated in the State of Pará, Brazil Resistência Antimicrobiana de Shigella spp. isoladas no Estado do Pará

INTRODUCTION: Shigella spp. are Gram-negative, nonsporulating, rod-shaped bacteria that belong to the family Enterobacteriaceae and are responsible for shigellosis or bacillary dysentery, an important cause of worldwide morbidity and mortality. METHODS: We studied the antibiotic resistance profiles of 122 Shigella spp. strains (81 S. flexneri, 41 S. sonnei, 1 S. boydii) isolated from patients (female and male from 0 to 80 years of age) presenting diarrhea in different districts of the State of Pará, in the North of Brazil. The antibiotic resistance of the strains, isolated from human fecal samples, was determined by the diffusion disk method and by using the VITEK-2 system. RESULTS: The highest resistance rate found was the resistance rate to tetracycline (93.8%), followed by the resistance rate to chloramphenicol (63.9%) and to trimethoprim/sulfamethoxazole (63.1%). Resistance to at least three drugs was more common among S. flexneri than S. sonnei (39.5% vs. 10%). Six (4.9%) strains were susceptible to all the antibiotics tested. All strains were susceptible to cefotaxime, ceftazidime, ciprofloxacin, nalidixic acid and nitrofurantoin. CONCLUSIONS: High rates of multidrug resistance in Shigella spp. are a serious public health concern in Brazil. It is extremely important to continuously monitor the antimicrobial resistances of Shigella spp. for effective therapy and control measures against shigellosis.INTRODUÇÃO: Shigella spp. são bactérias Gram-negativas, não esporuladas, em forma de bastonete, pertencentes a família Enterobacteriaceae responsáveis pela shigelose ou disenteria bacilar, uma importante causa de mortalidade e morbidade mundial. MÉTODOS: Foi estudado o perfil de resistência a antimicrobianos de 122 amostras de Shigella spp. (81 S. flexneri, 41 sonnei, 1 S. boydii) isoladas de pacientes (sexo feminino e masculino com faixa etária de 0 a 80 anos) com distúrbios gastrointestinais em diferentes municípios no Estado do Pará, Brasil. A resistência antimicrobiana das amostras isoladas de coprocultura, foi determinada pelo método de difusão em disco e pelo sistema Vitek II. RESULTADOS: A maior resistência foi observada em relação à tetraciclina (93,8%), seguida de cloranfenicol (63,9%), e trimetoprimsulfametoxazol (63,1%). Multirresistência a pelo menos três antimicrobianos foi mais comum em S. flexneri comparada a S. sonnei (39,5% vs. 10%). Seis (4,9%) amostras foram sensíveis a todos antimicrobianos testados. Todas as amostras apresentaram sensibilidade a cefotaxima, ceftazidima, ciprofloxacina, ácido nalidixico e nitrofurantoína. CONCLUSÕES: As altas taxas de multirresistência de Shigella spp. são um sério problema de saúde pública no Brasil. Sendo assim, torna-se extremamente importante um monitoramento contínuo da resistência antimicrobiana de Shigella spp. para uma terapia efetiva e medidas de controle contra shigelose

Characterization of antimicrobial resistance of samples of Shigella spp. isolated in the City of Belém, Pará State, Brazil (1990-2000)

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Pará. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Avaliou-se a resistência antimicrobiana de amostras de Shigella spp. isoladas de membros da população com faixa etárias de 6 meses a 81 anos, habitantes da Cidade de Belém, Estado do Pará, no período de 1990 a 2000. Foram analisadas 50 amostras de Shigella spp. identificadas no Laboratório de Enteroinfecções Bacterianas da Seção de Bacteriologia e Micologia do Instituto Evandro Chagas e mantidas na bacterioteca da referida seção. Na caracterização fenotípica, 32 (64%) dos isolados foram identificados como S. flexneri e 18 (36%) como S. sonnei. Resistência a cefalotina, cefazolina, cefuroxima, cefuroxima axetil e tobramicina foi observada em 100% das amostras, embora elas apresentassem 100% de sensibilidade a cefpodoxima, ceftriaxona, levofloxacina e norfloxacina. Das amostras, 2% apresentaram resistência a ticarcilina/ácido clavulânico, 8% a cefoxitina e 44% a ticarcilina. Os resultados obtidos evidenciaram a acentuada resistência a diversos antimicrobianos desta enterobactéria causadora de graves infecções em humanos.We evaluated the antimicrobial resistance of Shigella spp. samples isolated from inhabitants of the City of Belém, Pará State, Brazil, aged between 6 months and 81 years, from 1990 to 2000. We analyzed 50 samples of Shigella spp. that had been identified in the Laboratory of Enterobacterial Infections at the Bacteriology and Mycology Section of the Instituto Evandro Chagas and stored in its bacteria bank. After the phenotypic characterization, 32 (64%) isolates were identified as S. flexneri and 18 (36%) as S. sonnei. Resistance to cephalothin, cefazolin, cefuroxime, cefuroxime axetil and tobramycin was observed in 100% of the samples. However, they presented 100% susceptibility to cefpodoxime, ceftriaxone, levofloxacin and norfloxacin. Of the samples, 2% showed resistance to ticarcillin/clavulanic acid, 8% to cefoxitin and 44% to ticarcillin. The results showed a marked resistance to various antibiotics in these enterobacteria that cause serious infections in humans

Frequency of Shigella flexneri serotypes isolated in the state of Pará, Brazil (1979-2009)

Submitted by Sandra Infurna ([email protected]) on 2017-05-18T11:53:42Z No. of bitstreams: 1 ernesto2_hofer_etal_IOC_2012.pdf: 545644 bytes, checksum: b2142c5b7cdf3195c28da58c7715b21d (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2017-05-18T12:03:11Z (GMT) No. of bitstreams: 1 ernesto2_hofer_etal_IOC_2012.pdf: 545644 bytes, checksum: b2142c5b7cdf3195c28da58c7715b21d (MD5)Made available in DSpace on 2017-05-18T12:03:11Z (GMT). No. of bitstreams: 1 ernesto2_hofer_etal_IOC_2012.pdf: 545644 bytes, checksum: b2142c5b7cdf3195c28da58c7715b21d (MD5) Previous issue date: 2012Instituto Evandro Chagas. Departamento de Bacteriologia e Micologia. Ananindeua, PA, Brasil / Universidade Federal do Pará. Programa de Pós-Graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, Brasil.Instituto Evandro Chagas. Departamento de Bacteriologia e Micologia. Ananindeua, PA, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ. Brasil.The genus Shigella has been one of the most common causes of diarrhea in underdeveloped countries, being responsible for the mortality rate and morbidity in high risk populations such as children under five and the elderly. The aim of this study, developed at the Evandro Chagas Institute, Pará, Brazil, from 1979 to 2009, was to evaluate the frequency of species and serotypes of 122 isolates of Shigella spp. isolated from patients with acute diarrhea. The isolates were identified by bacterial culture media with selective indicators and biochemical tests, and serogroups and serotypes were determined by the slide agglutination test. The serogroups most frequently found were S. flexneri (66.4%) and S. sonnei (32.8%). The serotype 2a of S. flexneri was the most frequent (54.3%) followed by 1b (17.2%). Most patients were aged between 0-5 years (44.6%), of which 38.2% cases had S. flexneri and 47.5% S. sonnei. Patients older than 18 years accounted for 39.2% of infections, with 37.0% S. flexneri cases, 32.5% S. sonnei, and 100% S. boydii. These results emphasize the need for continuous surveillance in the State of Pará