n-n--n---triacetylfusarinine-c and Disease-Models--Animal

Siderophores represent important microbial virulence factors and infection biomarkers. Their monitoring in fermentation broths, bodily fluids, and tissues should be reproducible. Similar isolation, characterization, and quantitation studies can often have conflicting results, and without proper documentation of sample collection, data processing, and analysis methods, it is difficult to reexamine the data and reconcile these differences. In this Springer Nature Protocol, we present the procedure optimized for ferricrocin/triacetylfusarinine C extraction from biological material as well as for tissue fixation and cryosectioning for optical microscopy and for both elemental and molecular mass spectrometry imaging. Special attention is paid to siderophore data mining from conventional and product ion mass spectra, liquid chromatography, and mass spectrometry imaging datasets, performed here by our free software called CycloBranch.

Topics: Animals; Aspergillus fumigatus; Biomarkers; Chromatography, Liquid; Cryoultramicrotomy; Data Mining; Datasets as Topic; Disease Models, Animal; Ferric Compounds; Ferrichrome; Humans; Hydroxamic Acids; Invasive Pulmonary Aspergillosis; Mass Spectrometry; Rats; Siderophores; Software; Tissue Fixation

(68)Ga-triacetylfusarinine C ((68)Ga-TAFC) and (68)Ga-ferrioxamine E ((68)Ga-FOXE) showed excellent targeting properties in Aspergillus fumigatus rat infection model. Here, we report on the comparison of specificity towards different microorganisms and human lung cancer cells (H1299).. The in vitro uptake of (68)Ga-TAFC and (68)Ga-FOXE was studied in various fungal, bacterial and yeast cultures as well as in H1299 cells. The in vivo imaging was studied in fungal and bacterial rat infection and inflammation models.. (68)Ga-TAFC and (68)Ga-FOXE showed rapid uptake in A. fumigatus cultures, significantly lower in other fungal species and almost no uptake in other microorganisms and H1299 cells, except for (68)Ga-FOXE in Staphylococcus aureus. (68)Ga-TAFC and (68)Ga-FOXE revealed rapid uptake in the lungs of A. fumigatus-infected rats, low accumulation in sterile inflammation and no uptake in bacterial abscess.. We have shown that (68)Ga-FOXE and (68)Ga-TAFC have high uptake in A. fumigatus both in vitro and in vivo. (68)Ga-TAFC showed higher specificity, while (68)Ga-FOXE showed higher sensitivity.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; Cell Line, Tumor; Disease Models, Animal; Ferric Compounds; Fluorodeoxyglucose F18; Gallium Radioisotopes; Humans; Hydroxamic Acids; Peptides, Cyclic; Positron-Emission Tomography; Rats; Rats, Inbred Lew; Siderophores