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All boys underwent allogeneic HSCT, 5 boys from among matched unrelated donors, and two from among matched sibling donors. The mean age at the time of HSCT for patients with X-ALD was 8.14 years (ranging from 5 to 10 years). Sibling donors were evaluated for VLCFA levels and accepted only if the acid levels remained within the normal range. Two patients (1 and 3) were subjected to transplantation twice because of graft rejection. Conditioning regimens were as follows: busulfan (Bu)/cyclophosphamide (Cy)-based in 2 cases, treosulfan (Treo)/fludarabine (Flu)/Cy-based in 3 cases, and Treo/Cy-based in other cases. Acute Graft versus Host Disease (aGvHD) prophylaxis included either antithymocyte globulin (ATG) plus [http://www.selleckchem.com/products/AZD0530.html check details] cyclosporine A (CSA)/methotrexate (Mtx) �C in HSCT from unrelated donors or ATG/CSA �C in HSCT from matched related donors. The clinical characteristics of the study group are shown in Table 1. Table 1 The clinical characteristics of the study group. The study was conducted in accordance with the guidelines of the local University Ethics Committee for conducting research involving humans. Parents or legal guardians of all patients provided their signed informed consent to participate in the examination. Blood samples Blood samples of 1�C2 mL were taken from patients after a 12-hour overnight [http://en.wikipedia.org/wiki/Diglyceride diglyceride] fast. The next step was centrifugation for 4 minutes at 3000 g. The serum was separated and stored frozen at ?20��C before the analysis [10]. VLCFA analysis Analysis of VLCFA levels, as the methyl esters, was carried out in accordance with the method described by Stradomska TJ [10]. Serum VLCFA levels (C26:0, C24:0, C22:0) were determined by a gas chromatography technique. Fat acids were esterified with a mixture of methanol and acetyl chloride. The fatty acid methyl esters were extracted in the next step with hexane and separated by capillary column gas chromatography using a Hewlett Packard 5890 series II GC equipped with a capillary column (OV-1 30 m, 0.25ID). The ionization detector was flamed. The programmed [http://www.selleckchem.com/products/GDC-0941.html GDC-0941 concentration] separation included an initial temperature of 140��C for 1 minute, followed by an incremental increase of 8��C/min to 280��C for 10 minutes. Concentrations of VLCFA were calculated with HP-Chem-Station Software (Hewlett Packard) with tricosanoic acid as an internal standard. Identification of the fatty acid methyl esters was performed by using commercially available fatty acid standards (Sigma Chemical Co, St. Louis, Mo., USA) [10]. MR imaging protocol MR imaging was performed on a 1.5 T Signa Hdx scanner (GE Healthcare, Medical System) using a 16-channel coil dedicated for head and spine imaging. In 2 boys MRI examinations were performed under general anesthesia due to their young age (5 and 7 years). All children underwent plain MR imaging before and after HSCT.