Epidemiological and clinical profile of infective endocarditis at a Brazilian tertiary care center: an eight-year prospective study

Abstract INTRODUCTION: Infective endocarditis (IE) is a systemic infectious disease requiring a multidisciplinary team for treatment. This study presents the epidemiological and clinical data of 73 cases of IE in Rio de Janeiro, Brazil. METHODS This observational prospective cohort study of endocarditis patients during an eight-year study period described 73 episodes of IE in 70 patients (three had IE twice). Community-associated (CAIE) and healthcare-acquired infective endocarditis (HAIE) were diagnosed according to the modified Duke criteria. The collected data included demographic, epidemiologic, and clinical characteristics, including results of blood cultures, echocardiographic findings, surgical interventions, and outcome. RESULTS: Analysis of data from the eight-year study period and 73 cases (70 patients) of IE showed a mean age of 46 years (SD=2.5 years; 1-84 years) and that 65.7% were male patients. The prevalence of CAIE and HAIE was 32.9% and 67.1%, respectively. Staphylococcus aureus (30.1%), Enterococcus spp. (19.1%), and Streptococcus spp. (15.0%) were the prevalent microorganisms. The relevant signals and symptoms were fever (97.2%; mean 38.6 + 0.05°C) and heart murmur (87.6%). Vegetations were observed in the mitral (41.1%) and aortic (27.4%) valves. The mortality rate of the cases was 47.9%. CONCLUSIONS: In multivariate analysis, chronic renal failure (relative risk [RR]= 1.60; 95% confidence interval [CI] 1.01-2.55), septic shock (RR= 2.19; 95% CI 1.499-3.22), and age over 60 years (RR= 2.28; 95% CI 1.44-3.59) were indirectly associated with in-hospital mortality. The best prognosis was related to the performance of cardiovascular surgery (hazard ratio [HR]= 0.51; 95% CI 0.26-0.99).

Improved sealing ability promoted by calcium silicate-based root canal sealers

Aim: This study investigated the sealing ability produced by AH Plus (Dentsply DeTrey, Konstaz, Germany), EndoSequence BC sealer (Brasseler USA, Savannah, GA, USA), GuttaFlow (Coltène/Whaledent, Altstätten, France) and MTA Fillapex (Angelus, Londrina, Brazil). Methdos: A total of forty-six single-root human canines were prepared and randomly divided into four experimental groups (n=10): MTA Fillapex, EndoSequence BC Sealer, AH Plus or GuttaFlow. Teeth with intact crowns served as negative controls (n=3) and teeth filled with only gutta-percha served as positive controls (n=3). Teeth were mounted in a two-chamber apparatus and exposed to Enterococcus faecalis. The number of days over a 60-days period was recorded for the appearance of turbidity in the lower chamber. Kaplan-Meier method was used to estimate the survival curves. The nonparametric log-rank test was used to compare the survival curves using a significance level of 0.05. Results: The results at the end of the observation time were further analyzed by chi-square testing. All positive controls leaked within 24 h, whereas none of the negative controls leaked after 60 days. EndoSequence BC Sealer and MTA Fillapex had significant less bacterial leakage (P<0.05) than the other tested sealers. No significant difference between AH Plus and GuttaFlow was observed (P>0.05). Conclusion: In conclusion, calcium-silicate based root canal sealers promoted improved sealing ability when compared to other endodontic sealers

Genome sequence of a multidrug-resistant Corynebacterium striatum isolated from bloodstream infection from a nosocomial outbreak in Rio de Janeiro, Brazil

Multidrug-resistant (MDR) Corynebacterium striatum has been cited with increased frequency as pathogen of nosocomial infections. In this study, we report the draft genome of a C. striatum isolated from a patient with bloodstream infection in a hospital of Rio de Janeiro, Brazil. The isolate presented susceptibility only to tetracycline, vancomycin and linezolid. The detection of various antibiotic resistance genes is fully consistent with previously observed multidrug-resistant pattern in Corynebacterium spp. A large part of the pTP10 plasmid of MDR C. striatum M82B is present in the genome of our isolate. A SpaDEF cluster and seven arrays of CRISPR-Cas were found.

Cytotoxicity of three light-cured resin cements on 3T3 fibroblasts / Citotoxicidade de três cimentos resinosos fotopolimerizáveis sobre fibroblastos 3T3

Introduction: Light-cured resin cements are the first choice for the cementation of laminate veneers. Ideally, they should be biocompatible and offer minimum risks to patients. Objective: The aim of this study was to evaluate, in vitro, the cytotoxicity of three resin cements: Variolink II, Ivoclar Vivadent (C1), Allcem Veneer, FGM (C2), and Rely X Veneer, 3M ESPE (C3). Material and method: Twenty four samples of each of the cements were fabricated in a standardized metal mold, light activated, and transferred to a 96-well cell plate with culture of fibroblasts. After 24, 48, and 72h of incubation, cytotoxicity was assessed and cell viability was calculated by the methyl-thiazol-tetrazolium (MTT) colorimetric assay. Absorbance was measured at 570 nm using a microplate spectrophotometer. Result: The following results were found: Variolink II presented viability of 72.24% (SD 6.80) after 24h, 83.92% (SD 5.26) after 48h, and 92.77% (SD 5.59) after 72h; Allcem Veneer exhibited viability of 70.46% (SD 12.91) after 24h, 85.03% (SD 21.4) after 48h, and 70.46% (SD 12.91) after 72h; Rely X Veneer showed viability of 5.06% (SD 0.88) after 24h, 5.84% (SD 1.18) after 48h, and 6.99% (SD 1.34) after 72h. Conclusion: Under these testing conditions, Rely X Veneer presented significantly higher cytotoxicity compared with those of the other light-cured resin cements assessed.

Introdução: Cimentos resinosos fotopolimerizáveis são materiais de eleição para a cimentação de facetas laminadas. Devem ser biocompatíveis oferencendo riscos mínimos ao uso clínico em pacientes. Objetivo: O objetivo desse trabalho foi avaliar in vitro a citotoxicidade de três cimentos resinosos: Variolink II (Ivoclar Vivadent), Allcem Veneer, (FGM) e Rely X Veneer (3M ESPE). Material e método: Vinte e quatro corpos de prova de cada cimento foram confeccionados em matrizes metálicas padronizadas e inseridos em placa de cultura de células de noventa e seis poços contendo fibroblastos da linhagem 3T3. As células foram cultivadas em meio de cultivo celular RPMI 1640 com 5% de soro fetal bovino, com 0,1% de penicilina/estreptomicina em estufa a 37◦C, em atmosfera úmida com 5% de CO2. O grau de citotoxicidade de cada cimento foi avaliado após os tempos de contato de 24h, 48h e 72h através do método MTT (3-(4,5-dimetiltiazol-2yl)-2,5- difenil brometo de tetrazolina), que avalia a viabilidade celular pela função mitocondrial. Após os tempos estabelecidos, as amostras foram removidas, tratadas e levadas ao espectofotômetro de microplaca para leitura da absorbância em 570nm. Resultado: O cimento Variolink apresentou em 24h viabilidade de 72,24% (±6,80), em 48h de 83,92% (± 5,26) e de 92,77% (±5,59) em 72h. Allcem Veneer apresentou viabilidade de 70,46% (± 12,91) em 24h; 85,03% (± 21,4) em 48h e 70,46% (± 12,91) em 72h. O RelyX Veneer demonstrou viabilidade de 5,06% (± 0,88) em 24h; 5,84% (± 1,18) em 48h e 6,99% (± 1,34) em 72h. Conclusão: Estes resultados demonstraram que o cimento Rely-X se apresentou significativamente mais citotóxico nas condições testadas.

Sphygmomanometers and thermometers as potential fomites of Staphylococcus haemolyticus: biofilm formation in the presence of antibiotics

BACKGROUND The association between Staphylococcus haemolyticus and severe nosocomial infections is increasing. However, the extent to which fomites contribute to the dissemination of this pathogen through patients and hospital wards remains unknown. OBJECTIVES In the present study, sphygmomanometers and thermometers were evaluated as potential fomites of oxacillin-resistant S. haemolyticus (ORSH). The influence of oxacillin and vancomycin on biofilm formation by ORSH strains isolated from fomites was also investigated. METHODS The presence of ORSH on swabs taken from fomite surfaces in a Brazilian hospital was assessed using standard microbiological procedures. Antibiotic susceptibility profiles were determined by the disk diffusion method, and clonal distribution was assessed in pulsed-field gel electrophoresis (PFGE) assays. Minimum inhibitory concentrations (MICs) of oxacillin and vancomycin were evaluated via the broth microdilution method. Polymerase chain reaction (PCR) assays were performed to detect the mecA and icaAD genes. ORSH strains grown in media containing 1/4 MIC of vancomycin or oxacillin were investigated for slime production and biofilm formation on glass, polystyrene and polyurethane catheter surfaces. FINDINGS ORSH strains comprising five distinct PFGE types were isolated from sphygmomanometers (n = 5) and a thermometer (n = 1) used in intensive care units and surgical wards. ORSH strains isolated from fomites showed susceptibility to only linezolid and vancomycin and were characterised as multi-drug resistant (MDR). Slime production, biofilm formation and the survival of sessile bacteria differed and were independent of the presence of the icaAD and mecA genes, PFGE type and subtype. Vancomycin and oxacillin did not inhibit biofilm formation by vancomycin-susceptible ORSH strains on abiotic surfaces, including on the catheter surface. Enhanced biofilm formation was observed in some situations. Moreover, a sub-lethal dose of vancomycin induced biofilm formation by an ORSH strain on polystyrene. MAIN CONCLUSIONS Sphygmomanometers and thermometers are fomites for the transmission of ORSH. A sub-lethal dose of vancomycin may favor biofilm formation by ORSH on fomites and catheter surfaces.

Efetividade do ácido peracético na desinfecção rápida de cones de guta-percha e de Resilon expostos ao Enterococcus faecalis / Effectiveness of peracetic acid in rapid disinfection of gutta-percha and Resilon cones exposed to Enterococcus faecalis

Objetivo: o objetivo desse trabalho foi verificar a efetividade do ácido peracético a 2% na descontaminação rápida de cones de guta-percha e de Resilon, comparado ao hipoclorito de sódio e à clorexidina. Métodos: os cones de guta-percha e Resilon foram imersos por cinco minutos para contaminação em suspensão de Enterococcus faecalis, e divididos em grupos (n = 10): ácido peracético a 2% (um e três minutos); NaOCl a 5,25% (um e três minutos); clorexidina a 2% (um e três minutos). Após a realização dos protocolos em teste, os cones foram transferidos para tubos de ensaio contendo meio Enterococcosel® e, então, foram mantidos em estufa a 37º por 48 horas. Após o período de observação, os tubos foram avaliados e os que apresentam turvação do meio foram consideradas positivos. Resultados: os resultados demonstraram que o ácido peracético a 2% parece ser efetivo para a descontaminação de ambos os tipos de cone, de forma alternativa ao NaOCl a 5,25%, enquanto a clorexidina a 2% apresentou menor efetividade (p < 0,05). Conclusão: Ambas as soluções, ácido peracético a 2% e NaOCl a 5,25%, foram efetivas na descontaminação de cones de guta-percha ou de Resilon nos tempos testados.

Corynebacterium diphtheriae putative tellurite-resistance protein (CDCE8392_0813) contributes to the intracellular survival in human epithelial cells and lethality of Caenorhabditis elegans

Corynebacterium diphtheriae, the aetiologic agent of diphtheria, also represents a global medical challenge because of the existence of invasive strains as causative agents of systemic infections. Although tellurite (TeO3^2-) is toxic to most microorganisms, TeO3^2--resistant bacteria, including C. diphtheriae, exist in nature. The presence of TeO3^2--resistance (Te^R) determinants in pathogenic bacteria might provide selective advantages in the natural environment. In the present study, we investigated the role of the putative Te^R determinant (CDCE8392_813gene) in the virulence attributes of diphtheria bacilli. The disruption of CDCE8392_0813 gene expression in the LDCIC-L1 mutant increased susceptibility to TeO3^2- and reactive oxygen species (hydrogen peroxide), but not to other antimicrobial agents. The LDCIC-L1 mutant also showed a decrease in both the lethality of Caenorhabditis elegansand the survival inside of human epithelial cells compared to wild-type strain. Conversely, the haemagglutinating activity and adherence to and formation of biofilms on different abiotic surfaces were not regulated through the CDCE8392_0813 gene. In conclusion, the CDCE8392_813 gene contributes to the Te^R and pathogenic potential of C. diphtheriae.

Biofilm production by multiresistant Corynebacterium striatum associated with nosocomial outbreak

Corynebacterium striatum is a potentially pathogenic microorganism that causes nosocomial outbreaks. However, little is known about its virulence factors that may contribute to healthcare-associated infections (HAIs). We investigated the biofilm production on abiotic surfaces of multidrug-resistant (MDR) and multidrug-susceptible (MDS) strains of C. striatum of pulsed-field gel electrophoresis types I-MDR, II-MDR, III-MDS and IV-MDS isolated during a nosocomial outbreak in Rio de Janeiro, Brazil. The results showed that C. striatum was able to adhere to hydrophilic and hydrophobic abiotic surfaces. The C. striatum 1987/I-MDR strain, predominantly isolated from patients undergoing endotracheal intubation procedures, showed the greatest ability to adhere to all surfaces. C. striatum bound fibrinogen to its surface, which contributed to biofilm formation. Scanning electron microscopy showed the production of mature biofilms on polyurethane catheters by all pulsotypes. In conclusion, biofilm production may contribute to the establishment of HAIs caused by C. striatum.

Clonal multidrug-resistant Corynebacterium striatum within a nosocomial environment, Rio de Janeiro, Brazil

Corynebacterium striatum is a potentially pathogenic microorganism with the ability to produce outbreaks of nosocomial infections. Here, we document a nosocomial outbreak caused by multidrug-resistant (MDR) C. striatum in Rio de Janeiro, Brazil. C. striatum identification was confirmed by 16S rRNA and rpoB gene sequencing. Fifteen C. striatum strains were isolated from adults (half of whom were 50 years of age and older). C. striatum was mostly isolated in pure culture from tracheal aspirates of patients undergoing endotracheal intubation procedures. The analysis by pulsed-field gel electrophoresis (PFGE) indicated the presence of four PFGE profiles, including two related clones of MDR strains (PFGE I and II). The data demonstrated the predominance of PFGE type I, comprising 11 MDR isolates that were mostly isolated from intensive care units and surgical wards. A potential causal link between death and MDR C. striatum (PFGE types I and II) infection was observed in five cases.

Diphtheria Antibodies and T lymphocyte Counts in Patients Infected with HIV-1

We assessed the IgG levels anti-diphtheria (D-Ab) and T cell counts (CD4+ and CD8+) in HIV-1 infected subjects undergoing or not highly active antiretroviral therapy (HAART). Approximately 70% of all HIV-1 patients were unprotected against diphtheria. There were no differences in D-Ab according to CD4 counts. Untreated patients had higher D-Ab (geometric mean of 0.62 IU/ml) than HAART-patients (geometric mean of 0.39 IU/ml). The data indicated the necessity of keeping all HIV-1 patients up-to-date with their vaccination.

Atividade antimicrobiana de materiais reparadores de uso endodôntico pelos testes de difusão em ágar e por contato direto / Antimicrobial activity of repair materials of endodontic use by the agar diffusion test and the direct contact

O presente trabalho teve como objetivo avaliar a atividade antimicrobiana dos materiais reparadores de uso endodôntico: MTA/Bio Angelus®, cimento Portland branco, MTA Angelus® branco e MTA ProRoot® branco sobre os micro-organismos Enterococcus faecalis, Escherichia coli e Staphylococcus aureus, pelos os testes de difusão em ágar e por contato direto. Os dados experimentais foram obtidos através da mensuração do halo de inibição formado e pela contagem de unidades formadoras de colônia, respectivamente. De posse dos resultados, foi possível verificar que o MTA/Bio Angelus® apresentou atividade antimicrobiana superior para todas as espécies bacterianas testadas tanto no teste de difusão em ágar quanto no teste por contato direto e que este último teste demonstrou ser uma metodologia segura, reproduzível e confiável.

The aim of this study was to evaluate the antimicrobial activity of repair materials of endodontic use MTA/Bio Angelus®, white Portland cement, white MTA Angelus® and white MTA ProRoot® on Enterococcus faecalis, Escherichia coli and Staphylococcus aureus by the agar diffusion test and the direct contact test. Data was retrieved by measuring the zones of inhibition and by counting colony-forming units, respectively. The results showed that MTA/Bio Angelus® presented superior antimicrobial activity against all bacteria in the agar diffusion test and in the direct contact test. In addition, our results suggest that the use of the direct contact test is a safe, reproducible and trustable method.

Aggregative adherent strains of Corynebacterium pseudodiphtheriticum enter and survive within HEp-2 epithelial cells

Corynebacterium pseudodiphtheriticum is a well-known human pathogen that mainly causes respiratory disease and is associated with high mortality in compromised hosts. Little is known about the virulence factors and pathogenesis of C. pseudodiphtheriticum. In this study, cultured human epithelial (HEp-2) cells were used to analyse the adherence pattern, internalisation and intracellular survival of the ATCC 10700 type strain and two additional clinical isolates. These microorganisms exhibited an aggregative adherence-like pattern to HEp-2 cells characterised by clumps of bacteria with a "stacked-brick" appearance. The differences in the ability of these microorganisms to invade and survive within HEp-2 cells and replicate in the extracellular environment up to 24 h post infection were evaluated. The fluorescent actin staining test demonstrated that actin polymerisation is involved in the internalisation of the C. pseudodiphtheriticum strains. The depolymerisation of microfilaments by cytochalasin E significantly reduced the internalisation of C. pseudodiphtheriticum by HEp-2 cells. Bacterial internalisation and cytoskeletal rearrangement seemed to be partially triggered by the activation of tyrosine kinase activity. Although C. pseudodiphtheriticum strains did not demonstrate an ability to replicate intracellularly, HEp-2 cells were unable to fully clear the pathogen within 24 h. These characteristics may explain how some C. pseudodiphtheriticum strains cause severe infection in human patients.

Difteria pelo Corynebacterium ulcerans: uma zoonose emergente no Brasil e no mundo / Corynebacterium ulcerans diphtheria: an emerging zoonosis in Brazil and worldwide / Difteria por Corynebacterium ulcerans: una zoonosis emergente en Brasil y en el mundo

O artigo revisa a literatura sobre a emergência de infecções humanas causadas por Corynebacterium ulcerans em diversos países, incluindo o Brasil. Foi realizada análise de artigos publicados entre 1926 e 2011 nas bases Medline/PubMed e SciELO, bem como artigos e informes do Ministério da Saúde. Apresenta-se um esquema de triagem, rápido, econômico e de fácil execução, capaz de permitir a realização do diagnóstico presuntivo de C. ulcerans e C. diphtheriae na maioria dos laboratórios brasileiros públicos e privados. A circulação de C. ulcerans em vários países, aliada aos recentes casos de isolamento do patógeno no Rio de Janeiro, é um alerta a clínicos, veterinários e microbiologistas sobre a ocorrência de difteria zoonótica e a circulação do C. ulcerans em regiões urbanas e rurais do território nacional e/ou da América Latina.

The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America.

El articulo revisa la literatura sobre la emergencia de infecciones humanas causadas por Corynebacterium ulcerans en diversos países, incluyendo Brasil. Se realizó análisis de artículos publicados entre 1926 y 2011 en las bases Medline/Pubmed y SciELO, así como artículos e informes del Ministerio Brasileño de la Salud. Se presenta un esquema de selección, rápido, económico y de fácil ejecución, capaz de permitir la realización del diagnóstico presuntivo de C. ulcerans y C. diphtheriae en la mayoría de los laboratorios brasileños públicos y privados. La circulación de C. ulcerans en varios países, aliada a los recientes casos de aislamiento del patógeno en Rio de Janeiro (Sureste de Brasil), es un alerta a clínicos, veterinarios y microbiólogos sobre la ocurrencia de difteria zoológica y la circulación de C. ulcerans en regiones urbanas y rurales del territorio nacional y/o de América Latina.

Isolamento e identificação de enterococcus sp em infecções endodônticas primárias / Isolation and identification of enterococcus sp in primary endodontic infections

Este estudo objetivou o isolamento e caracterização bioquímica de Enterococcus sp de canais radiculares portadores de necrose pulpar de 70 pacientes. Foram isoladas 35 amostras de Enterococcus sp em 50% dos pacientes (n = 35) com a seguinte prevalência: 33 amostras Enterococcus faecalis (94,28%), 1 amostra Enterococcus faecium (2,85%) e 1 amostra Enterococcus durans (2,85%). Dos dentes com cultura positiva, 20 apresentaram sintomatologia dolorosa. Odor fétido e secreção purulenta foram observados em três dentes (8,57%). Dos 35 pacientes com cultura positiva, quatro apresentaram edema no momento da coleta e apenas um paciente apresentou fístula. A espécie E. faecalis foi majoritária, estando relacionada aos sinais e sintomas.

A influência do peróxido de carbamida sobre a formação de colônias de estreptococos do grupo mutans / The influence of carbamide peroxide on the formation of colonies of mutans streptococci

Com o desenvolvimento da terapia clareadora, tornou-se necessária uma avaliação acerca da segurança de sua utilização com relação à microbiota oral. Este trabalho teve como objetivo verificar a influência de agentes clareadores contendo diferentes concentrações de peróxido de carbamida sobre a colonização de quatro amostras de estreptococos do grupomutans. Foram testados os géis clareadores contendo 10%, 15%, 20% e 35% de peróxido de carbamida, alémdos controles positivo (clorexidina 0,2%) e negativo (solução salina estéril). A maior parte das amostras apresentou sensibilidade superior se comparados ao controle positivo, sendo a resposta concentração dependente. Concluímos que o peróxido de carbamida modula negativamente a colonização por estreptococosdo grupo mutans em todas as concentrações testadas.

Efeitos do consumo diário de probiótico sobre a microbiota cariogênica / Effects of daily consumption of probiotic on the cariogenic microbiota

Probióticos são definidos como micro-organismos vivos que, quando administrados em quantidade adequada, podem trazer benefícios ao hospedeiro melhorando seu equilíbrio intestinal. Estudos recentes têm sugerido que alguns probióticos funcionam como um método auxiliar no controle da doença cárie, pois podem diminuir os níveis salivares de Streptococcusmutans. Muitas bactérias, incluindo os gêneros Lactobacillus e Bifidobacterium possuem propriedades probióticas. Diversos veículos podem ser utilizados para administrá-los, entretanto o mais comum é o iogurte. Porém mais estudos sobre probióticos são necessários para comprovar seus reais benefícios na cavidade oral, permitindo aos dentistas indicarem seu consumo com segurança.

Aspectos fenotípicos e do potencial patogênico de amostra de Corynebacterium diphtheriae isolada de processo de osteomielite em paciente com neoplasia / Phenotypic aspects and pathogenic potential of Corynebacterium diphtheriae strains isolated from cancer patient with osteomyelitis

Além da difteria permanecer endêmica em diversos países, os clínicos e microbiologistas também devem permanecer atentos ao fato de amostras atoxinogênicas de Corynebacterium diphtheriae causarem infecções invasivas, inclusive em pacientesimunocomprometidos e/ou hospitalizados. Um grupo de microrganismos, incluindo C. diphtheriae, tem sido relacionado com quadros de osteomielite. Em casos de câncer, pode ser favorecido o aparecimento de quadros de osteomielite em decorrência de contaminação por via hematogênica, foco infeccioso ou lesão contígua ao osso. Entretanto, ainda são poucas as investigações relativas ao potencial patogênico de cepas atoxinogênicas de C. diphtheriae. No presente estudo, foi descrito o primeiro caso de isolamento de C.diphtheriae subsp. mitis atoxinogênico e do biotipo não fermentador de sacarose (BR5015) de osteomielite em paciente com câncer.O microrganismo foi capaz de expressar os seguintes fatores de virulência: expressão de perfil de aderência misto dos tipos agregativo e difuso (AA-AD) e elevada (11,13%) capacidade de sobrevivência intracitoplasmática em células epiteliais humanas (HEp-2) além da produção de porfirina e de enzimas catalase, nitrato redutase e DNAse. C. diphtheriae atoxinogênico não deve serconsiderado como mero contaminante, uma vez que pode estar direta ou indiretamente relacionado com o estabelecimento e/ou manutenção de processos infecciosos de origens diversas, incluindo osteomielite.

As well diphtheria remaining endemic in several countries, clinicians and microbiologists must also remain alert to the fact that nontoxigenic samples of Corynebacterium diphtheriae are capable of causing invasive infections, especially in hospitalized and/or immunocompromised patients. Patients with cancer are more susceptible to the appearance of cases of osteomyelitisobtained by hematogenic contamination, an infectious focus or by lesions adjacent to bone. Many microorganisms may be related to cases of osteomyelitis, including C. diphtheriae. However, there are still only a low number of investigations into the pathogenic potential of nontoxigenic strains of C. diphtheriae. The present study is the first documented case of isolation of a nontoxigenic C.diphtheriae subsp. mitis of the non sucrose-fermenting biotype (BR5015 strain) from osteomyelitis in the frontal bone of a patient with adenoid cystic carcinoma. The virulence factors tests were as follows: expression of a mixed adherence patterns of aggregativediffuse(AA-DA) types; high (11.13%) ability to survive within HEp-2 cells; DNase, catalase, nitrate-reductase activities. Therefore, nontoxigenic C. diphtheriae should not be merely regarded as a contaminant, since it can be directly or indirectly related to the establishment and/or maintenance of infectious processes, including osteomyelitis.

Fibrinogen binds to nontoxigenic and toxigenic Corynebacterium diphtheriae strains

The production of fibrinous exudates may play an important role in determining the outcome of bacterial infection. Although pseudomembrane formation is a characteristic feature of diphtheria, little is known about the fibrinogen (Fbn)-binding properties of Corynebacterium diphtheriae strains and the influence of the gene that codes for diphtheria toxin (tox gene) in this process. In this study we demonstrated the ability of C. diphtheriae strains to bind to Fbn and to convert Fbn to fibrin. Bacterial interaction with rabbit plasma was evaluated by both slide and tube tests. Interaction of microorganisms with human Fbn was evaluated by both enzyme linked immunosorbent assay (ELISA) and fluorescein isothiocyanate-conjugated (FITC) Fbn binding assays. Nontoxigenic and toxigenic strains formed bacterial aggregates in the presence of plasma in the slide tests. The ability to convert Fbn to a loose web of fibrin in the plasma solution in the tube tests appeared to be a common characteristic of the species, including strains that do not carry the tox gene. Fbn binding to C. diphtheriae strains occurred at varying intensities, as demonstrated by the FITC-Fbn and ELISA binding assays. Our data suggest that the capacity to bind to Fbn and to convert Fbn to fibrin may play a role in pseudomembrane formation and act as virulence determinants of both nontoxigenic and toxigenic strains.

Cytotoxicity evaluation of two root canal sealers and a commercial calcium hydroxide paste on THP1 cell line by Trypan Blue assay

OBJECTIVE: The aim of this investigation was to evaluate the cytotoxicity of two brands of root canal sealers, epoxy-resin based and zinc oxide-eugenol based, and one commercial calcium hydroxide paste on a monocyte cell line THP-1. MATERIAL AND METHODS: Undiluted (crude extract) and diluted extracts to 10 percent, 1 percent, 0.1 percent, 0.01 percent, 0.001 percent and 0.0001 percent of the sealers were tested for cytotoxicity to THP-1 cells using the trypan blue assay. Extracts were obtained according to ISO standard. Data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests at 5 percent significance level. RESULTS: Crude extract of AH Plus and Fill Canal killed approximately 90 percent of THP-1 cells versus 36 percent of THP-1 cells killed by L&C crude extract (p<0.05). Ten-fold dilutions of L&C, Fill Canal and AH Plus killed 24, 35 and 61 percent of THP-1 cells (p<0.05), respectively. Dilutions lesser than 1 percent caused minimal cell death as compared to the control groups (p>0.05), except for L&C 1 percent extract. CONCLUSIONS: The results revealed that the L&C paste crude extract was less cytotoxic to THP-1 cells than AH Plus or Fill Canal crude extracts.