The Impact of Pseudomonas aeruginosa Infection in Adult Cystic Fibrosis Patients-A Single Polish Centre Study.

BACKGROUND: Pseudomonas aeruginosa (PA) is one of the most predominant pathogens of lung infections, often causing exacerbations in adult patients with cystic fibrosis (CF). MATERIALS AND METHODS: Microbiological characterization of 74 PA isolates and to evaluate the correlations between the bacterial features and 44 adult Polish CF cohort clinical parameters. RESULTS: The most common variant in the CF transmembrane conductance regulator (CFTR) gene was F508del (76.3%), followed by 3849+10kbC>T (26.3%). A total of 39.4% of the PA isolates showed multiple resistances. In patients with parameters pointing to a decline in lung function, there was a statistically significant moderate correlation with ß-lactam resistance and a weak correlation between hospital frequency and colistin resistance. The mucoidity did not correlate with the biofilm formation ability, which showed 41.9% of the isolates. Proteolytic activity, observed in 60.8% of the clinical isolates, was weakly associated with motility detected in 78.4% of the strains. The genetic profiles of the PA were highly heterogeneous, and a weak positive correlation was established between cluster group and biofilm formation. CONCLUSION: The findings suggest that there is a high variety in P. aeruginosa populations in adult CF patients. There is a need to monitor PA strains in groups of patients with cystic fibrosis, in particular, in terms of the occurrence of antibiotic resistance related to a decline in lung function.

Phenotypic and genotypic characteristics of Pseudomonas aeruginosa isolated from cystic fibrosis patients with chronic infections.

Patients with cystic fibrosis are predisposed to chronic respiratory tract infections caused by Pseudomonas aeruginosa. As the disease progresses, the microorganism diversifies into genotypically and phenotypically different strains which may coexist in the patient's airways for years. Adaptation of the microorganism to the airways of patients with cystic fibrosis probably occurs in response to the host's airway environment, the elements of the immune system and antibiotic therapy. Due to the chronic persistence of the microorganism in the airways, a comprehensive molecular analysis was conducted. The analysis included 120 strains isolated from 10 adult cystic fibrosis patients with chronic P. aeruginosa infection. The aim of the study was to analyze the molecular patterns of P. aeruginosa strains and to trace their transmission in the population of cystic fibrosis patients, as well as to study a relationship of the disease with specific phenotypic features. In the research, a genotypic analysis of P. aeruginosa was performed using pulsed-field gel electrophoresis. The results of a number of phenotypic features of the strains were added to the outcomes of the molecular studies. As a result, 28 different genotypes were distinguished. The study also showed cross-transmission of strains between patients. 3 transmissible clusters were identified, including IG1 and IG2 clusters with 9 strains of P. aeruginosa each, obtained from 2 patients and IG3 cluster with 6 strains of P. aeruginosa isolated from 3 patients. Moreover, it was found that in some patients, several unrelated strains of P. aeruginosa may transiently or permanently infect the respiratory tract. A comprehensive understanding of the P. aeruginosa adaptation may help to develop more effective antimicrobial therapies and to identify new targets for future drugs in order to prevent progression of the infection to chronic stages.

Main-chain flexibility and hydrophobicity of ionenes strongly impact their antimicrobial activity: an extended study on drug resistance strains and Mycobacterium.

The spread of antibiotic-resistant pathogens and the resurgence of tuberculosis disease are major motivations to search for novel antimicrobial agents. Some promising candidates in this respect are cationic polymers, also known as synthetic mimics of antimicrobial peptides (SMAMPs), which act through the membrane-lytic mechanism. Development of resistance toward SMAMPs is less likely than toward currently employed antibiotics; however, further studies are needed to better understand their structure-activity relationship. The main objective of this work is to understand the cross-influence of hydrophobicity, main-chain flexibility, and the topology of ionenes (polycations containing a cationic moiety within the main-chain) on activity. To fulfill this goal, a library of ionenes was developed and compared with previously investigated molecules. The obtained compounds display promising activity against the model microorganisms and drug-resistance clinical isolates, including Mycobacterium tuberculosis. The killing efficiency was also investigated, and results confirm a strong effect of hydrophobicity, revealing higher activity for molecules possessing the flexible linker within the polymer main-chain.

Influence of PEG Subunit on the Biological Activity of Ionenes: Synthesis of Novel Polycations, Antimicrobial and Toxicity Studies.

An alarming increase of antibiotic resistance among pathogens creates an urgent need to develop new antimicrobial agents. Many reported polycations show high antimicrobial activity along with low hemolytic activity. Unfortunately, most of those molecules remain highly cytotoxic against various mammalian cells. In this work, a systematic study on the impact of triethylene glycol monomethyl ether side groups (short polyethylene glycol (PEG) analog) on antimicrobial, hemolytic, and cytotoxic properties of novel amphiphilic ionenes is presented. A detailed description of synthesis, leading to well-defined alternating polymers, which differ in structural elements responsible for hydrophilicity (PEG) and hydrophobicity (alkyl chain), is presented. Obtained results show that the PEG moiety and fine-tuned hydrophilic-lipophilic balance of ionenes synergistically lead to low cytotoxic, low hemolytic molecules with high activity against S. aureus, including methicillin-resistant strains (MRSA). Additionally, the results of mechanistic studies on bacterial cells and fluorescently labeled liposomes are also discussed.

Molecular analysis of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients.

Pseudomonas aeruginosa is a severe bacterial pathogen. Due to the genetic flexibility among strains, chronic airways infection can lead to mortality among cystic fibrosis (CF) patients. It is essential to develop patient-specific therapy which will rely on phenotypic and genomic diversity. The primary objective of this study was to assess the genomic variability of P. aeruginosa strains, using two different molecular techniques for tracking the epidemiological transmissions. This study applied a multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) for an efficient genotyping of clinical P. aeruginosa strains isolated from CF patients and compared results with a TRS-PCR typing. The percentage similarity analysis was performed using the categorical multi-state coefficient and UPGMA method. Based on the MLVA and TRS-PCR group assessment, 43 P. aeruginosa strains/variants were detected among the 63 clinical isolates from eight CF patients. The study of P. aeruginosa isolates has revealed that during chronic bacterial infections, CF patients harbor different P. aeruginosa strains or variants within the same host over the years. P. aeruginosa genotypes diversity may result from infection with several strains and result from a microevolution process of an initially acquired strain. The TRS-PCR method proposed in this work can complement the MLVA scheme. It can also be used as a preliminary method for genetic typing of P. aeruginosa isolates in CF patients.

Biofilm Formation on Dental Implant Biomaterials by Staphylococcus aureus Strains Isolated from Patients with Cystic Fibrosis.

Implants made of ceramic and metallic elements, which are used in dentistry, may either promote or hinder the colonization and adhesion of bacteria to the surface of the biomaterial to varying degrees. The increased interest in the use of dental implants, especially in patients with chronic systemic diseases such as cystic fibrosis (CF), is caused by an increase in disease complications. In this study, we evaluated the differences in the in vitro biofilm formation on the surface of biomaterials commonly used in dentistry (Ti-6Al-4V, cobalt-chromium alloy (CoCr), and zirconia) by Staphylococcus aureus isolated from patients with CF. We demonstrated that S. aureus adherence and growth depends on the type of material used and its surface topography. Weaker bacterial biofilm formation was observed on zirconia surfaces compared to titanium and cobalt-chromium alloy surfaces. Moreover, scanning electron microscopy showed clear differences in bacterial aggregation, depending on the type of biomaterial used. Over the past several decades, S. aureus strains have developed several mechanisms of resistance, especially in patients on chronic antibiotic treatment such as CF. Therefore, the selection of an appropriate implant biomaterial with limited microorganism adhesion characteristics can affect the occurrence and progression of oral cavity infections, particularly in patients with chronic systemic diseases.

[Analysis of the frequency of isolation and drug resistance of microorganisms isolated from the airways of adult CF patients treated in the Institute of Tuberculosis and Lung Disease during 2008-2011]. / Analiza czestosci izolacji i profil lekoopornosci drobnoustrojów izolowanych z dróg oddechowych doroslych chorych na mukowiscydoze leczonych w Instytucie Gruzlicy i Chorób Pluc w latach 2008­2011.

INTRODUCTION: Cystic fibrosis (CF) is the most common genetic autosomal recessive genetic disease. The most serious symptoms are observed in the lungs. Recurrent respiratory infections are the main causes of the hospitalizations and deaths of cystic fibrosis patients. Pathogens that commonly infect the airways of adult CF patients include Staphylococcus aureus and Pseudomonas aeruginosa. The aim of this study was to analyse the microorganisms cultured from the airways of adult CF patients and to test the antimicrobial resistance of the most frequently isolated bacteria. MATERIAL AND METHODS: In this study, 1422 isolates of 89 CF patients were collected during a 4-year period. The microorganisms were cultured and identified according to standard microbiological procedures. Identification and drug susceptibility were performed in an automatic system Phoenix (BD), Vitek2Compact (bioMérieux), and disk-diffusion method by Kirby-Bauer. RESULTS: Among the 1422 strains the most frequent pathogens were Pseudomonas aeruginosa (55.6%) and Staphylococcus aureus (37.8%). A total of 482 (61.0%) strains of 790 isolates of P. aeruginosa, were identified as P. aeruginosa mucoid phenotype. The isolates with mucoid phenotypes were more susceptible to antibiotics than non-mucoid. Eighty-six strains of S. aureus showed resistance to methicillin (MRSA), which accounted for 16.0% of all strains of S. aureus. CONCLUSIONS: The analysis of microbiological materials from adult CF patients treated in IGiChP allowed the determination of the prevalence of potentially pathogenic microorganisms. The data obtained are consistent with the literature.