Detection of mixed microbial biofilms on central venous catheters removed from Intensive care Unit Patients

Cateteres venosos centrais inseridos em pacientes internados em unidade de terapia intensiva foram avaliados por métodos microbiológicos (cultura semi-quantitativa) e microscopia eletrônica de varredura a fim de detectar adesão microbiana e correlacionar com a cultura de sangue. Durante o período de estudo, foram avaliados 59 pacientes com cateter venoso central. A idade dos pacientes, sexo, sítio de inserção e tempo de permanência do cateter foram anotados. O cateter era de poliuretano não tunelizado e de único lúmen. O sangue para cultura foi coletado no momento da remoção do cateter. de 63 pontas de cateteres, 30 (47,6%) foram colonizadas e a infecção encontrada em 5 (23,8%) cateteres. A infecção foi mais prevalente em 26 pacientes (41,3%) com cateteres inseridos em veia subclávia do que nos 3 (3,2%) inseridos em veia jugular. A infecção foi observada com mais freqüência em cateteres com tempo de permanência maior do que sete dias. Os microrganismos isolados incluíram 32 estafilococos coagulase-negativa (29,7%), 61 bactérias Gram-negativas (52,9%), 9 estafilcocos coagulase-positiva (8,3%) e 3 leveduras (2,7%). Como agentes causais de infecções em unidade de terapia intensiva foram isolados E. aerogenes, P. aeruginosa, A. baumannii. Os antimicrobianos com maior atividade in vitro contra as bactérias Gram-negativas foram o imipenem e contra as Gram-positivas vancomicina, cefepime, penicilina, rifampicina e tetraciclina. As análises por microscopia eletrônica de varredura revelaram biofilmes sobre a superfície de todos os cateteres examinados.

Biossíntese e recentes avanços na produção de celulose bacteriana

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudos in silico da proteína AlkB e prospecção de bactérias alcanotróficas em solos da bacia petrolífera potiguar

Some microorganisms from virgin ecosystems are able to use petroleum it as source of carbon and energy. The knowledge of microbial biodiversity can help to reveal new metabolic systems for utilization alkanes with biotechnological importance. The aim of this study is: i) Accomplish an in silico study of the AlkB protein aimed to understand the probable mechanism involved on selectivity of alkanes in Gram positive and Gram negative bactéria. ii) prospect and analyze the response of the microbial alkanotrophics communities in soil and mangrove sediments of BPP RN and soil of Atlantic forest in the Horto Dois Irmãos Reserve area/PE using the molecular biomarker, gene alkB; with the PCR and PCR-DGGE approach

Bacterial sterilization by a dielectric barrier discharge (DBD) in air

Cold atmospheric plasma treatment of microorganisms and living tissues has become a popular topic in modern plasma physics and in medical science. The plasma is capable of bacterial inactivation and noninflammatory tissue modification, which makes it an attractive tool for treatment of skin diseases, open injuries and dental caries. Because of their enhanced plasma chemistry, Dielectric Barrier Discharges (DBDs) have been widely investigated for some emerging applications such as biological and chemical decontamination of media at ambient conditions. Despite the high breakdown voltage in air at atmospheric pressure, the average current of DBD discharges is low. Therefore, a DBD can be applied in direct contact with biological objects without causing any damage. In this work a 60 Hz DBD reactor, which generates cold atmospheric plasma inside Petri dishes with bacterial culture, is investigated. Samples of Staphylococcus aureus, a Gram-positive bacterium and Escherichia coil a Gram-negative bacterium were selected for this study. The bacterial suspensions were evenly spread on agar media planted in Petri dishes. The reactor electrodes were placed outside the Petri dish, thus eliminating the risk of samples microbial contamination. The covered Petri dish with agar medium in it serves as dielectric barrier during the treatment. The plasma processing was conducted at same discharge power (similar to 1.0 W) with different exposure time. Sterilization of E. coil and S. aureus was achieved for less than 20 min. Plasma induced structural damages of bacteria were investigated by Scanning Electron Microscopy. (C) 2010 Elsevier B.V. All rights reserved.

TLR4 and CD14 receptors expressed in rat pineal gland trigger NFKB pathway

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do caráter oxidante da violaceína

Violacein is a violet pigment isolated from many gram-negative bacteria, especially from Chromobacterium violaceum, a betaproteobacterium found in the Amazon River in Brazil. It has potential medical applications as an antibacterial, fungicide, anti-tryptanocidal, anti-ulcerogenic and anti-cancer drug, among others. Furthermore, its pro-oxidant activity has been suggested, but only in two specific tumor lineages. Thus, in the present study, the prooxidant effects of violacein were investigated in both normal and tumor cells, seeking to evaluate the cell responses. The evaluation of violacein cytotoxicity using the Trypan blue dye exclusion method indicated that CHO-K1 cells were more resistant than tumor HeLa cells. The oxidative stress induced by violacein was manifested as an increase in intracellular SOD activity in CHO-K1 and MRC-5 cells at a specific concentration range. Nevertheless, a decrease was detected specifically at 6-12 μM in HeLa and MRC-5 cells. Interestingly, the increase in SOD activity was not followed by a concomitant increase in catalase activity. Regarding to oxidative stress biomarkers, increased protein carbonylation and lipid hydroperoxides levels were detected respectively in CHO-K1 and MRC-5 cells treated with violacein at 1.5-3 μM and 3 μM, which may be an evidence that this compound causes oxidative stress specifically in these conditions. Additionally, it is believed that the decline in cell viability observed in MRC-5 cells and HeLa treated with violacein at 6-12 M is due to mechanisms not related to oxidative stress. Moreover, the results suggested that violacein might cause oxidative stress by increasing endogenous levels of O2 -, since the occurrence of an expressive change in SOD activity. In addition, in order to evaluate the antioxidant activity of violacein in the absence of a biological system, the total antioxidant and iron chelating activity were evaluated, so that antioxidant activities were detected at 30 and 60 μM of violacein. Altogether, the results indicate that although oxidative stress is triggered by incubation with violacein, it did not seem to be high enough to cause serious damage to cell biomolecules in HeLa cells and only at specific concentrations in CHOK-1 and MRC-5 cells. Comparing the results obtained in cell culture and the in vitro antioxidant activity evaluation, the results confirmed that violacein presents opposing oxidant features when in presence or absence of a biological system and the antioxidant character only occurs at high concentrations of the pigment.

Clinical and laboratory evaluation of horses after intraperitoneal injection of lipopolysaccharide (LPS)

Horses are very sensitive to Gram-negative bacterial lipopolysaccharides (LPS) which, when introduced into the blood stream, induce several responses mediated by endogenous pro-inflammatory mediators originating from bacteriolysis or granulocyte disintegration. Intraperitoneal LPS injection was used to mimic the proposed route of endotoxin travel in clinical cases. The clinical signs and laboratory findings demonstrated a dose- dependent effect and were more consistently observed with 500 ng/kg of LPS. We conclude that intraperitoneal injection of LPS produces the same endotoxic status as obtained by systemic injection of LPS, even during the initial phase.

Occurrence of yeasts, pseudomonads and enteric bacteria in the oral cavity of patients undergoing head and neck radiotherapy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Subgingival microbiota from Cebus apella (capuchin monkey) with different periodontal conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antimicrobial resistance of aerobes and facultative anaerobes isolated from the oral cavity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antigenicity of Primary Endodontic Infection against Macrophages by the Levels of PGE(2) Production

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antigenic Activity of Bacterial Endodontic Contents from Primary Root Canal Infection with Periapical Lesions against Macrophage in the Release of Interleukin-1 beta and Tumor Necrosis Factor alpha

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A dominant function of p38 mitogen-activated protein kinase signaling in receptor activator of nuclear factor-kappa B ligand expression and osteoclastogenesis induction by Aggregatibacter actinomycetemcomitans and Escherichia coli lipopolysaccharide

Background and Objective: Lipopolysaccharide from gram-negative bacteria is one of the microbial-associated molecular patterns that initiate the immune/inflammatory response, leading to the tissue destruction observed in periodontitis. The aim of this study was to evaluate the role of the p38 mitogen-activated protein kinase (MAPK) signaling pathway in lipopolysaccharide-induced receptor activator of nuclear factor-kappa B ligand (RANKL) expression by murine periodontal ligament cells.Material and Methods: Expression of RANKL and osteoprotegerin mRNA was studied by reverse transcription-polymerase chain reaction upon stimulation with lipopolysaccharide from Escherichia coli and Aggregatibacter actinomycetemcomitans. The biochemical inhibitor SB203580 was used to evaluate the contribution of the p38 MAPK signaling pathway to lipopolysaccharide-induced RANKL and osteoprotegerin expression. Stable cell lines expressing dominant-negative forms of MAPK kinase (MKK)-3 and MKK6 were generated to confirm the role of the p38 MAPK pathway. An osteoclastogenesis assay using a coculture model of the murine monocytic cell line RAW 264.7 was used to determine if osteoclast differentiation induced by lipopolysaccharide-stimulated periodontal ligament was correlated with RANKL expression.Results: Inhibiting p38 MAPK prior to lipopolysaccharide stimulation resulted in a significant decrease of RANKL mRNA expression. Osteoprotegerin mRNA expression was not affected by lipopolysaccharide or p38 MAPK. Lipopolysaccharide-stimulated periodontal ligament cells increased osteoclast differentiation, an effect that was completely blocked by osteoprotegerin and significantly decreased by inhibition of MKK3 and MKK6, upstream activators of p38 MAPK. Conditioned medium from murine periodontal ligament cultures did not increase osteoclast differentiation, indicating that periodontal ligament cells produced membrane-bound RANKL.Conclusion: Lipopolysaccharide resulted in a significant increase of RANKL in periodontal ligament cells. The p38 MAPK pathway is required for lipopolysaccharide-induced membrane-bound RANKL expression in these cells.

Clinical and microbiological studies of children and adolescents receiving orthodontic treatment

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Seasonal effect on Brazilian propolis antibacterial activity

The behavior of microorganisms towards the antibiotic action of propolis has been widely investigated. Since reports dealing with seasonal effect on propolis activity are not available, this assay was carried out aiming to observe the in vitro antimicrobial activity of propolis, collected during the four seasons, on bacterial strains isolated from human infections. Dilution of ethanolic extract of propolis (EEP) in agar was the method performed, with serial concentrations ranging from 0.4 to 14.0% (% v/v). The behavior of some bacteria was analysed according to the incubation period in medium plus propolis, and the survival curve was plotted. It was verified that the growth of Gram-positive bacteria is inhibited by low propolis concentrations (0.4%) whereas Gram-negative bacteria were less susceptible to this substance, the minimal inhibitory concentration ranging from 4.5 to 8.0%. There was no significant difference with regards to the seasonal effect on the survival curve of Staphylococcus aureus and Escherichia coli; after incubation with propolis, there was an efficient antimicrobial action, mainly towards Gram-positive bacteria. (C) 2000 Elsevier B.V. Ireland Ltd. All rights reserved.