Gene regulatory network inference and analysis of multidrug-resistant Pseudomonas aeruginosa

BACKGROUND Healthcare-associated infections caused by bacteria such as Pseudomonas aeruginosa are a major public health problem worldwide. Gene regulatory networks (GRN) computationally represent interactions among regulatory genes and their targets. They are an important approach to help understand bacterial behaviour and to provide novel ways of overcoming scientific challenges, including the identification of potential therapeutic targets and the development of new drugs. OBJECTIVES The goal of this study was to reconstruct the multidrug-resistant (MDR) P. aeruginosa GRN and to analyse its topological properties. METHODS The methodology used in this study was based on gene orthology inference using the reciprocal best hit method. We used the genome of P. aeruginosa CCBH4851 as the basis of the reconstruction process. This MDR strain is representative of the sequence type 277, which was involved in an endemic outbreak in Brazil. FINDINGS We obtained a network with a larger number of regulatory genes, target genes and interactions as compared to the previously reported network. Topological analysis results are in accordance with the complex network representation of biological processes. MAIN CONCLUSIONS The properties of the network were consistent with the biological features of P. aeruginosa. To the best of our knowledge, the P. aeruginosa GRN presented here is the most complete version available to date.

Development of an experimental model of neutrophilic pulmonary response induction in mice

BACKGROUND: Several lung diseases are characterized by a predominantly neutrophilic inflammation. A better understanding of the mechanisms of action of some drugs on the airway inflammation of such diseases may bring advances to the treatment. OBJECTIVE: To develop a method to induce pulmonary neutrophilic response in mice, without active infection. METHODS: Eight adult Swiss mice were used. The study group (n = 4) received an intranasal challenge with 1 x 10(12) CFU/ml of Pseudomonas aeruginosa (Psa), frozen to death. The control group (n = 4) received an intranasal challenge with saline solution. Two days after the intranasal challenge, a bron¡choalveolar lavage (BAL) was performed with total cell and differential cellularity counts. RESULTS: The total cell count was significantly higher in the group with Psa, as compared to the control group (median of 1.17 x 10(6) and 0.08 x 10(6), respectively, p = 0.029). In addition to this, an absolute predominance of neutrophils was found in the differential cellularity of the mice that had received the Psa challenge. CONCLUSIONS: The model of inducing a neutrophilic pulmonary disease using frost-dead bacteria was successfully developed. This neutrophilic inflammatory response induction model in Swiss mice lungs may be an important tool for testing the anti-inflammatory effect of some antimicrobial drugs on the inflammation of the lower airways.

Imunidade humoral em pacientes com fibrose cística / Humoral immunity of patients with cystic fibrosis

Pacientes com FC frequentemente såo acometidos por infecçöes pulmonares, principalmente por P. aeruginosa tanto da variedade nåo mucóide como mucóide. Esta última, quando se instala, é acompanhada geralmente de deterioraçåo progressiva da funçåo pulmonar. Detectam-se nestes pacientes, anticorpos para o microorganismo e diversos produtos por ele produzidos. A imnunoeletroferese cruzada tem sido tradicionalmente utilizada para pesquisa de precipitinas para P. aeruginosa. Esta surgem pouco após o início do episódio infeccioso e guardam correlaçåo com a cronicidade do processo, quando se mostram mais numerosoas. Em pacientes infectados por P. auruginosa mucóide, o número de precipitinas costuma ser maior que nos infectados pela variedade nåo-mucóide. As precipitinas habitualmente nåo surgem nos pacientes apenas colonizados, servindo assim para distinguir casos de colonizaçåo dos de infecçåo, e indicando a oportunidade do emprego de antibioticoterapia nestes pacientes. O número de precipitinas se reduz após controle do processo infeccioso. Outras técnicas vêm sendo desenvolvidas com sucesso, para a pesquisa de anticorpos para P. aureginosa, visando a obtençåo de maior sensibilidade e especificidade, abrindo caminho para novas possibilidades de investigaçåo