Purpose With the increase of especially hospital-acquired infections, timely and accurate diagnosis of bacterial infections is crucial for effective patient care. Molecular imaging has the potential for specific and sensitive detection of infections. Siderophores are iron-specific chelators recognized by specific bacterial transporters, representing one of few fundamental differences between bacterial and mammalian cells. Replacing iron by gallium-68 without loss of bioactivity is possible allowing molecular imaging by positron emission tomography (PET). Here, we report on the preclinical evaluation of the clinically used siderophore, desferrioxamine-B (Desferal®, DFO-B), radiolabelled with 68 Ga for imaging of bacterial infections. Methods In vitro characterization of [ 68 Ga]Ga-DFO-B included partition coefficient, protein binding and stability determination. Specific uptake of [ 68 Ga]Ga-DFO-B was tested in vitro in different microbial cultures. In vivo biodistribution was studied in healthy mice and dosimetric estimation for human setting performed. PET/CT imaging was carried out in animal infection models, representing the most common pathogens. Results DFO-B was labelled with 68 Ga with high radiochemical purity and displayed hydrophilic properties, low protein binding and high stability in human serum and PBS. The high in vitro uptake of [ 68 Ga]Ga-DFO-B in selected strains of Pseudomonas aeruginosa , Staphylococcus aureus and Streptococcus agalactiae could be blocked with an excess of iron-DFO-B. [ 68 Ga]Ga-DFO-B showed rapid renal excretion and minimal retention in blood and other organs in healthy mice. Estimated human absorbed dose was 0.02 mSv/MBq. PET/CT images of animal infection models displayed high and specific accumulation of [ 68 Ga]Ga-DFO-B in both P. aeruginosa and S. aureus infections with excellent image contrast. No uptake was found in sterile inflammation, heat-inactivated P. aeruginosa or S. aureus and Escherichia coli lacking DFO-B transporters. Conclusion DFO-B can be easily radiolabelled with 68 Ga and displayed suitable in vitro characteristics and excellent pharmacokinetics in mice. The high and specific uptake of [ 68 Ga]Ga-DFO-B by P. aeruginosa and S. aureus was confirmed both in vitro and in vivo, proving the potential of [ 68 Ga]Ga-DFO-B for specific imaging of bacterial infections. As DFO-B is used in clinic for many years and the estimated rad...

Pseudomonas aeruginosa is an increasingly prevalent opportunistic pathogen that causes a variety of life-threatening nosocomial infections. Novel strategies for the development of new antibacterial treatments as well as diagnostic tools are needed. One of the novel diagnostic strategies for the detection of infection could be the utilization of siderophores. Siderophores are low-molecular-weight chelators produced by microbes to scavenge essential iron. Replacing iron in siderophores by suitable radiometals, such as Ga-68 for positron emission tomography (PET) imaging, opens approaches for targeted imaging of infection. Here we report on pyoverdine PAO1 (PVD-PAO1), a siderophore produced by P. aeruginosa, labelled with Ga-68 for specific imaging of Pseudomonas infections. PVD-PAO1 was labelled with Ga-68 with high radiochemical purity. The resulting complex showed hydrophilic properties, low protein binding and high stability in human serum. In vitro uptake of 68Ga-PVD-PAO1 was highly dependent on the type of microbial culture. In normal mice 68Ga-PVD-PAO1 showed rapid pharmacokinetics with urinary excretion. PET imaging in infected animals displayed specific accumulation of 68Ga-PVD-PAO1 in infected tissues and better distribution than clinically used 18F-fluorodeoxyglucose (18F-FDG) and 68Ga-citrate. Ga-68 labelled pyoverdine PAO1 seems to be a promising agent for imaging of P. aeruginosa infections by means of PET.

Pichia fabianii, a yeast rarely causing human infections, was isolated from the blood of a patient with aortic valve endocarditis. The isolates were initially identified biochemically as Candida pelliculosa, but based on direct sequencing of the ITS2 region of rRNA, they were subsequently reidentified as P. fabianii. Antifungal therapy with fluconazole and later with voriconazole led to the development of resistant variants which had high MIC values to both antifungals. Strong biofilm formation by this yeast could also have played a role in the development of its resistance and allowed for its persistence on the infected valve during antifungal therapy. To our knowledge, this is the first published case of endocarditis and the fourth human infection caused by this yeast species.

A rather fast and complicated progression of an infection caused by some strains of Staphylococcus aureus could be associated with the expression and co-action of virulence factor complexes in these strains. This study screened the antibiotic susceptibility and prevalence of virulence markers in isolates of meticillin-resistant S. aureus (MRSA) obtained from patients hospitalized at the University Hospital in Olomouc, Czech Republic. A total of 100 isolates was screened for 13 genes encoding extracellular virulence determinants (tst, pvl, eta, etb, sea, seb, sec, sed, see, seg, seh, sei and sej) and for their distribution in sample types. Eighty-nine isolates were positive for at least one of the genes. Genes for etb, pvl, see and seh were not detected in any of the MRSA isolates. No statistically significant differences in the occurrence of the determinants studied among sample types were found. INTRODUCTIONStaphylococcus aureus is one of the major nosocomial pathogens. Particular attention should be paid to meticillin-resistant S. aureus (MRSA). Resistance to meticillin is determined by the presence of the mecA gene encoding penicillin-binding protein with very low affinity to blactam antibiotics (Chambers, 1997). S. aureus produces a broad spectrum of extracellular and cell wall-associated virulence determinants (Foster, 2002). Among them, a wide variety of surface adhesins known as microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) has been described (Patti et al., 1994).The most common staphylococcal proteins anchored in the cell wall are proteins with affinity to fibrinogen (i.e. clumping factors A and B, encoded by the clfA and clfB genes, respectively), fibronectin (fnbA), collagen (cna), sialoprotein (bbp), elastin (ebpS) and adhesins with unknown function (sdrC and sdrE) (Jonsson et al., 1991;Josefsson et al., 1998; McDevitt et al., 1997;Ní Eidhin et al., 1998; Park et al., 1996;Speziale et al., 1986;Tung et al., 2000). The second group of virulence factors is represented by a family of bacterial proteins with superantigen activity: enterotoxins A-E, G-R and U (encoded by the genes seasee, seg-ser and seu), toxic shock syndrome toxin-1 (TSST-1, encoded by tst), exfoliative toxins A and B (eta and etb) and other toxins such as a-, b-, c-and d-toxin and the Panton-Valentine leukocidin (pvl) (Arbuthnott et al., 1982;Bhakdi & Tranum-Jensen, 1991;Bohach et al., 1990;Prevost et al., 1995).Apart from syndromes caused by toxin production, S. aureus pathogenesis results from synergistic interactions of a variety of the above-mentioned factors. Exfoliative toxin and pyrogenic toxin superantigen production enables S. aureus to cause staphylococcal scalded skin syndrome (SSSS), staphylococcal toxic shock syndrome and staphylococcal food poisoning. Experimental models indicate that the expression of receptors for fibrinogen and fibronectin is associated with endocarditis, whereas the presence of adhesins for sialoprotein, collagen and fibronectin is associated with arthritis and osteomyel...