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1

Differences in metabolic profiles of planktonic and biofilm cells in Staphylococcus aureus-1H Nuclear Magnetic Resonance search for candidate biomarkers

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Junka A. F., Deja S., Smutnicka D., Szymczyk P., Ziolkowski G., Bartoszewicz M., Mlynarz P.
Acta Biochimica Polonica
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2013
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tom 60
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nr 4
Staphylococcus aureus is responsible for many types of infections related to biofilm presence. As the early diagnostics remains the best option for prevention of biofilm infections, the aim of the work presented was to search for differences in metabolite patterns of S. aureus ATCC6538 biofilm vs. free-swimming S. aureus planktonic forms. For this purpose, Nuclear Magnetic Resonance (NMR) spectroscopy was applied. Data obtained were supported by means of Scanning Electron Microscopy, quantitative cultures and X-ray computed microtomography. Metabolic trends accompanying S. aureus biofilm formation were found using Principal Component Analysis (PCA). Levels of isoleucine, alanine and 2,3-butanediol were significantly higher in biofilm than in planktonic forms, whereas level of osmoprotectant glycine-betaine was significantly higher in planktonic forms of S. aureus. Results obtained may find future application in clinical diagnostics of S. aureus biofilm-related infections.
2

The application of impedance microsensors for real-time analysis of Pseudomonas aeruginosa biofilm formation

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Chabowski K., Junka A. F., Szymczyk P., Piasecki T., Sierakowskii A., Maczynska B., Nitsh K.
Polish Journal of Microbiology
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2015
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tom 64
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nr 2
Biofilms formed by nosocomial pathogens represent a major threat to patients undergoing invasive procedures. As prophylaxis remains the most efficient anti-biofilm option, it is of paramount importance to develop diagnostic tools able to detect biofilm at the early stage of formation. The present study investigates the ability of impedance microsensors to detect Pseudomonas aeruginosa biofilm presence using the impedance spectroscopy method. The measured data were analyzed using Electrical Equivalent Circuit modelling (EEC). It allowed to recognize conduction and polarization phenomena on the sensors surface and in its environment. The impedance assay results, confirmed by means of electron microscopy and quantitative cultures, indicate that specific EEC parameters may be used for monitoring the development of pseudomonal biofilm.
3

Use of the real time xCelligence system for purposes of medical microbiology

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Junka A. F., Janczura A., Smutnicka D., Maczynska B., Secewicz A., Nowicka J., Bartoszewicz M., Gosciniak G.
Polish Journal of Microbiology
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2012
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tom 61
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nr 3
Roche’s xCelligence impedance-measuring instrument is one of a few commercially available systems of such type. According to the best knowledge of authors, instrument was tested so far only for eukaryotic cell research. The aim of this work was to estimate xCELLigence suitability for the microbiological tests, including (i) measurement of morphological changes in eukaryotic cells as a result of bacterial toxin activity, (ii) measurement of bacterial biofilm formation and (iii) impact of antiseptics on the biofilm structure. To test the influence of bacterial LT enterotoxin on eukaryotic cell lines, Chinese Hamster Ovary (CHO) cell line and reference strain Escherichia coli ATTC 35401 were used. To investigate Roche’s instrument ability to measure biofilm formation and impact of antiseptics on its development, Staphylococcus aureus ATTC6538 reference strain was used. The data generated during the experiments indicate excellent ability of xCelligence instrument to detect cytopathic effect caused by bacterial LT endotoxin and to detect staphylococcal biofilm formation. However, interpretation of the results obtained during real-time measurement of antiseptic’s bactericidal activity against staphylococcal biofilm, caused many difficulties. xCelligence instrument can be used for real-time monitoring of morphological changes in CHO cells treated with bacterial LT enterotoxin and for real-time measurement of staphylococcal biofilm formation in vitro. Further investigation is necessary to confirm suitability of system to analyze antiseptic’s antimicrobial activity against biofilm in vitro.
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