Analysis of the human sera samples using GC-TOF-MS yielded 144 known metabolic structures and a total of 354 molecular features (Dataset).
Thus, in line with previous steroid-abuse rat studies [36 39], the present study was designed with a daily dose of 5.0 mg/kg for 6 consecutive days, followed by analysing hair, urine and sera samples using newly developed LC-MS/MS methods.
Together, these validations showed that the method is valid for the quantitation of human sera samples using calibration curves in serum, as well as using calibration curves in plasma (Tandem Labs 1999, 2001a, 2001b).
Then, we isolated the total RNA from each pooled sera sample using TRIzol reagent (Invitrogen) according to the manufacturer's instructions with minor modifications.
We stained all cells with appropriate dye-labeled Ig, mouse sera, or lamprey plasma and analyzed samples using a LSR II flow cytometer and fitted data to a one-site binding hyperbola model using PRISM software.
Viral DNA was extracted from 200 μL sera per sample using an AccuPrep Genomic DNA Extraction Kit (Bioneer, Korea) or QIAamp MinElute Virus Spin (Qiagen, Germany) as instructed in the manufacturer's manuals.
IgG, IgG1 and IgG2a antibodies in sera samples were determined using an indirect ELISA assay according to the manufacture's instructions (R&D Systems, USA).
When the generalised and combinational ROC curves were applied to the samples using sera from the healthy women as the controls these curves were not as informative as some of the individual features (see Figure 4B) but in these latter cases, only a small number of sera were positive (see Figure 3A).
Poly(A -enriched mRNA -enrichedied fromRNAe total RNA samples using Sera-mega Oligo(dT) beads (Illumina) and fragmented wash divalent cations at elevated tempurified.
Using the reagents of the mRNA Sample Prep Kit and in accordance with the RNA sequencing protocol provided by Illumina, poly(A)+ mRNA was selected out from the total RNA samples using Sera-mag magnetic oligo dT) beads.
