DocumentCode :
2472831
Title :
Development of a high-throughput monitoring technique of bacteria photodynamic inactivation
Author :
Cunha, Bernardo ; Sampaio, Pedro N. ; Calado, Cecilia R. C.
Author_Institution :
Dept. de Eng. Quim., Inst. Super. de Eng. de Lisboa, Lisbon, Portugal
fYear :
2015
fDate :
26-28 Feb. 2015
Firstpage :
1
Lastpage :
2
Abstract :
Summary form only given. Bacterial infections and the fight against them have been one of the major concerns of mankind since the dawn of time. During the `golden years´ of antibiotic discovery, during the 1940-90s, it was thought that the war against infectious diseases had been won. However currently, due to the drug resistance increase, associated with the inefficiency of discovering new antibiotic classes, infectious diseases are again a major public health concern. A potential alternative to antibiotic treatments may be the antimicrobial photodynamic inactivation (PDI) therapy. To date no indication of antimicrobial PDI resistance development has been reported. However the PDI protocol depends on the bacteria species [1], and in some cases on the bacteria strains, for instance Staphylococcus aureus [2]. Therefore the development of PDI monitoring techniques for diverse bacteria strains is critical in pursuing further understanding of such promising alternative therapy. The present works aims to evaluate Fourier-Transformed-Infra-Red (FT-IR) spectroscopy to monitor the PDI of two model bacteria, a gram-negative (Escherichia coli) and a gram-positive (S. aureus) bacteria. For that a high-throughput FTIR spectroscopic method was implemented as generally described in Scholz et al. [3], using short incubation periods and microliter quantities of the incubation mixture containing the bacteria and the PDI-drug model the known bactericidal tetracationic porphyrin 5,10,15,20-tetrakis (4-N, N, Ntrimethylammoniumphenyl)-porphyrin p-tosylate (TTAP4+). In both bacteria models it was possible to detect, by FTIR-spectroscopy, the drugs effect on the cellular composition either directly on the spectra or on score plots of principal component analysis. Furthermore the technique enabled to infer the effect of PDI on the major cellular biomolecules and metabolic status, for example the turn-over metabolism. In summary bacteria PDI was monitored in an economic, rapid (in minutes- , high-throughput (using microplates with 96 wells) and highly sensitive mode resourcing to FTIR spectroscopy, which could serve has a technological basis for the evaluation of antimicrobial PDI therapies efficiency.
Keywords :
Fourier transform infrared spectroscopy; antibacterial activity; biochemistry; biomedical measurement; cellular biophysics; drugs; microorganisms; molecular biophysics; patient monitoring; photodynamic therapy; principal component analysis; 5,10,15,20-tetrakis (4-N, N, Ntrimethylammoniumphenyl)-porphyrin p-tosylate; Escherichia coli; Fourier transformed infrared spectroscopy; PDI effect; PDI protocol; PDI-drug model content; S. aureus; Staphylococcus aureus; TTAP4+; alternative therapy; antibiotic class; antibiotic discovery; antibiotic treatment; antimicrobial PDI resistance development; antimicrobial PDI therapy efficiency evaluation; antimicrobial photodynamic inactivation; bacteria content; bacteria photodynamic inactivation; bacteria strain variation; bacterial infection; bacterial species dependence; bacterial strain dependence; bactericidal tetracationic porphyrin; cellular biomolecule; cellular composition; drug effect detection; drug resistance; economic rapid high-throughput bacteria PDI monitoring; gram negative bacteria; gram positive bacteria; high-throughput FTIR spectroscopy; high-throughput PDI monitoring technique development; incubation mixture composition; incubation mixture microliter quantity; infectious disease; metabolic status; microplate; model bacteria PDI monitoring; principal component analysis score plot; public health concern; short incubation period; turn-over metabolism; Antibiotics; Biomedical engineering; Immune system; Microorganisms; Monitoring; Spectroscopy; Antibiotic resistance; Infra-red spectroscopy; Photodynamic therapy;
fLanguage :
English
Publisher :
ieee
Conference_Titel :
Bioengineering (ENBENG), 2015 IEEE 4th Portuguese Meeting on
Conference_Location :
Porto
Type :
conf
DOI :
10.1109/ENBENG.2015.7088827
Filename :
7088827
Link To Document :
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