This is to mimic cases (e.g. soil microbes) where many species are not individually culturable.
By first removing Y chromosome values and then randomly omitting a varying number of values for autosomes, while retaining all X chromosome values, we could mimic cases with different percentage of gain.
Titres were even detectable at t = 0 (1.2 1.5 BU mL−1 in the haemophilia-mimic case and 0.6 1.6 BU in the factors-mixture case).
Figure 2 shows detailed results of residual FVIII C at t = 0 of the haemophilia-mimic case with three inhibitor dilutions.
Inhibitor curves were similar for the haemophilia-mimic case and the factors-mixture case in which FVIII was allowed to bind to VWF prior to contact with the inhibitor.
There were no discrepancies between the chromogenic and the one-stage clotting assay results at the 2 h time point of the inhibitory assay in the haemophilia-mimic case.
Kinetic results from the haemophilia-mimic case (i.e. addition of the FVIII concentrate to previously mixed VWF + inhibitor) showed that the BU titres of the inhibitor pool against pdFVIII/VWF were null at t = 0 and increased over time in a pattern almost identical to that of normal plasma.
The inhibitor titre for the pdFVIII/VWF complex concentrates was comparable to the titre in normal plasma at all time points studied in both the haemophilia-mimic case (in which FVIII is added to previously mixed VWF+inhibitor) and the factors-mixture case (in which the inhibitor is added to previously mixed VWF+FVIII).
In the haemophilia-mimic case, serial dilutions of inhibitor IgG were mixed with an equal volume of 2 IU mL−1 VWF and later incubated with an equal volume of 1 IU mL−1 of FVIII of different origin (pdFVIII/VWF, pdFVIII, rFVIII, BDD-rFVIII) and normal human plasma.
Conversely, the inhibitor titres obtained with all isolated FVIII products (pdFVIII, rFVIII and BDD-rFVIII) in the haemophilia-mimic case were significantly higher than in normal plasma and the pdFVIII/VWF complex concentrates at all the time points (P < 0.05 0.001), with ratios reaching 1.4 1.9 fold, respectively, after 2 h of incubation (titres ranging from 16.5 ± 2.1 to 23.3 ± 3.9 BU).
In an effort to verify the specificity of the SLC44A1- PRKCA fusion, FISH analysis was also performed on fifteen PGNTs mimics cases (cases 5 to 19) and demonstrated that none of these cases displayed the SLC44A1-PRKCA fusion (Fig. 3e and f).
