List of identified proteins from GeLC-MS/MS analysis of size exclusion chromatography fraction 2.

a<p>UniprotKB accession number.</p>b<p>Bands in which the protein was identified.</p>c<p>Percentage content according to label free quantitation approach.</p

SDS-PAGE separation of sheep HCF.

<p>Two lung (lanes 1–2, 5–6) and two liver (lanes 3–4, 7–8) HCF samples were separated under non-reducing (N, lanes 1–4) and reducing conditions (R, lanes 5–8) by SDS-PAGE on 15% acrylamide gels. Each sample was loaded after concentration and desalting by ultrafiltration. Due to the different starting protein concentrations, different volumes of each sample, ranging from 7.5 µL to 17.5 µL, where used to reach loads of 20 µg/lane. Coomassie Blue staining was performed. Lane M: molecular weight markers.</p

SDS-PAGE and relative protein abundance of fraction 2 from size exclusion chromatography of sheep HCF.

<p>A: Laemmli buffer with (R) or without 400 mM DTT (N) was added to aliquots of 500 ng (lanes 1N-1R), 1 µg (lanes 2N-2R) and 2 µg (lanes 3N-3R) of size exclusion chromatography fraction 2, and subjected to SDS-PAGE on 4–10% acrylamide gel. Lane M: molecular weight markers. Gel was silver stained and then analyzed by GeLC-MS/MS, by cutting lanes 3N and 3R in 7 bands (a–g) each. B: pie chart representing the relative protein abundance of fraction 2 from GeLC-MS/MS analysis of lane 3R. According to label free quantitation, Ag5 represents the most abundant component of the fraction, reaching about 63% of the total content.</p

Comparative evaluation of Ag5 and commercial ELISA kits.

<p>Boxplots summarizing the absorbance values obtained with the Ag5 preparation and commercial ELISA kits on CE patients and control subjects. Patient and control sera are plotted according to the clinical status. The dashed and dotted lines indicate the upper and lower cutoff values, respectively. ABS: absorbance at the wavelength required for each assay. Differences in absorbance between the patient and control groups were statistically significant for all the assays (P<0.001).</p

List of identified proteins from selected SDS-PAGE bands of Figure 1.

a<p>UniprotKB accession number.</p

Western immunoblotting of human sera against Ag5 enriched preparation.

<p>Fraction 2 from size exclusion chromatography of HCF was used as antigen in western blotting experiments under non-reducing conditions. Sera from CE patients (1–13) and control subjects (14–24) were tested against a total of 300 ng of protein sample loaded in each single-well gel; this amount corresponds approximately to about 10 ng of proteins per multiscreen slot. All CE patients sera react against the Ag5 protein band, although with a variable intensity probably depending on the antibody titer of each serum. Moreover, control sera do not give any non-specific response. All sera were tested in at least three separate experiments. The molecular weight region of Ag5 (60 kDa) is indicated on the left.</p

Summary of the ELISA results obtained with the Ag5 enriched antigen test and four commercial kits.

<p>Summary of the ELISA results obtained with the Ag5 enriched antigen test and four commercial kits.</p

Immunoreactive bands subjected to mass spectrometry analysis.

<p>HCF was separated by SDS-PAGE and the resulting gel was cut in two parts, one of them subjected to western blot experiment with serum 1, the other part stained and further processed for protein identification Left: Immunoreactive profile observed by western immunoblotting on non-reduced (N) and reduced (R) HCF samples. Right: reactive bands excised from the SDS-PAGE gel and subjected to LC-MS/MS identification. M: molecular weight markers.</p

Echinococcosis HDRs distribution by region.

<p>Echinococcosis HDRs distribution by region.</p

Direct costs of CE patients grouped by NUTS 1 for Italians and foreign patients in the study period.

<p>Direct costs of CE patients grouped by NUTS 1 for Italians and foreign patients in the study period.</p