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The ACI technique consisted of a two-step procedure, as originally described by Brittberg [1].
Sterile conditions were maintained during the whole blood culture procedure, as originally described [20].
If the biopsy is negative for metastatic disease, the patient undergoes the sentinel node procedure as originally planned.
The IAP was measured in a non-invasive manner through the aspiration port of a standard indwelling bladder catheter, in a modification of the procedure as originally described by Kron et al. The IAP was measured twice daily until discharge from the ICU or until there was no further need of a bladder catheter (i.e. a bladder catheter was not left in place for the sole purpose of measuring IAP).
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But limitations to the procedure as it was originally proposed have proven difficult to overcome for forensic samples.
The transplantation procedure was carried out as originally described [18, 19, 20]: all mice received general anaesthesia; laparotosternotomy was performed in donor mice to explant the graft; thereon lung grafts were stored in a preservation solution at 4 °C until transplant.
For an outgroup, we mapped unassembled reads of the closely related Asiatic honeybee A. cerana, available from the NCBI Short Read Archive (SRX339508) to the A. mellifera reference genome following the same procedure as the experiment that originally generated the data [ 28].
All the experimental procedures were carried out as originally proposed by Kodama et al [ 4], except that those authors injected a lower dose of alloxan (200 mg/kg of body weight).
We did not use generalised estimating equations (GEE) as originally intended [ 34] since this procedure was less suitable to the present outcomes than logistic regression analysis.
No 'batch effect' modification is performed during this procedure as the 70 gene x 559 patient dataset was that used to originally develop the GEP70 algorithm.
Although we lost the CTCs of two samples during the DEPArray™ procedure (as expected from the transfer efficiency), we found more CTC events than originally detected with CellSearch® in three cases.
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