Multivariate statistical analysis of the MR spectra.

<p>Loadings plots (on PC1) obtained by performing the PCA comparisons on the MR spectra of control and one treatment per analysis (as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0026155#pone-0026155-g003" target="_blank">Fig. 3</a>) acquired on polar extracts of PC3 (black line) and LNCaP (red line) prostate cancer cells following 48 hours of treatment with (A) LY294002 or (B) 17AAG. Enlarged sections of the loadings plots represent the region of 1.9–4.1 ppm. Ala: alanine; Asn: asparagine; Cho: choline; Cit: citrate; Cre: creatine; Fum: fumarate; Glc: glucose; Gln: glutamine; Gly: glycine; GPcho: glycerophosphocholine; GSH: glutathione; His: histidine; Ile: isoleucine; Lac: lactate; Leu: leucine; m-Ino: myo-inositol; Pcho: phosphocholine; Pcre: phosphocreatine; Phe: phenylalanine; Tau: taurine; Tyr: tyrosine; Val: valine, UDPS: UDP sugars.</p

Inhibition of target signaling pathways in cancer cells following drug treatment.

<p>Schematic of signaling pathways targeted by LY294002 and 17AAG and Western blots showing modulation of p-4E-BP1 and c-Raf (β-actin as loading control) levels following administration of DMSO (solvent control; C), LY294002 (L) or 17AAG (A).</p

Accumulation of citrate following treatment with 17AAG in PC3 prostate cancer cells.

<p>Enlarged section (2.49 – 2.72 ppm) of the MR spectra acquired on polar extracts of PC3 cells following 48 hours of treatment with DMSO (solvent control, black), LY294002 (green) and 17AAG (red). Spectra were normalized according to the probabilistic quotient normalization method.</p

Common metabolic changes in prostate and breast cancer cells following drug treatment.

<p>Quantification of selected metabolites (shown as percent of control, mean ± standard deviation) from MR spectra acquired on prostate (PC3 and LNCaP, N = 8) and breast (MCF-7, N = 3) cancer cell lines following 48 hours of treatment with (A) LY294002 or (B) 17AAG. *: p<0.05; **: p<0.005; ***: p<0.0005. Pcholine: phosphocholine.</p

Multivariate statistical analysis of the MR spectra.

<p>Scores plots (PC1 vs PC2) obtained by performing PCA on the MR spectra acquired on polar extracts (8 replicates per treatment condition) of PC3 and LNCaP prostate cancer cells. Control samples were compared to samples treated for 48 hours with either LY294002 ((A) for PC3 and (C) for LNCaP cells) or 17AAG ((B) for PC3 and (D) for LNCaP cells).</p

Additional file 7: of Serum metabolomic profiling predicts synovial gene expression in rheumatoid arthritis

Figure S6. Correlation between serum metabolites and synovial IL-1β and IL-8. (TIFF 14826 kb

Additional file 4: of Serum metabolomic profiling predicts synovial gene expression in rheumatoid arthritis

Figure S3. Correlation between serum metabolites and synovial CD19, CD79A, and IgGHC. (TIFF 14826 kb

Additional file 2: of Serum metabolomic profiling predicts synovial gene expression in rheumatoid arthritis

Figure S1. Correlation between synovial markers and serum metabolites. (TIFF 14826 kb

Hypoxia Triggers Major Metabolic Changes in AML Cells without Altering Indomethacin-Induced TCA Cycle Deregulation

Our previous studies have shown that the nonsteroidal anti-inflammatory drug indomethacin exhibits antileukemic activity <i>in vitro</i> and can inhibit the aldo-keto reductase AKR1C3, which we identified as a novel target in acute myeloid leukemia. However, the antileukemic actions of indomethacin are likely to be complex and extend beyond inhibition of either AKR1C3 or cycloxygenases. To further understand the antileukemic activity of indomethacin we have used untargeted nuclear magnetic resonance-based metabolic analysis to characterize the responses of KG1a and K562 cell lines in both normal culture conditions and in hypoxia, which better represents the tumor environment <i>in vivo</i>. Hypoxia induced dramatic metabolic changes in untreated KG1a and K562, including adaptation of both phospholipid and glycolytic metabolism. Despite these changes, both cell lines sustained relatively unaltered mitochondrial respiration. The administration of indomethacin induced similar metabolic responses regardless of the oxygen level in the environment. Notable exceptions included metabolites associated with <i>de novo</i> fatty acid synthesis and choline phospholipid metabolism. Collectively, these results suggest that leukemia cells have the inherent ability to tolerate changes in oxygen tension while maintaining an unaltered mitochondrial respiration. However, the administration of indomethacin significantly increased oxidative stress in both KG1a and K562, inducing mitochondrial dysfunction, regardless of the oxygenation conditions. These findings emphasize the particular pertinence of the tricarboxylic acid cycle to the survival of cancer cells and may explain why some antileukemic drugs have been discovered and developed successfully despite the use of culture conditions that do not reflect the hypoxic environment of cancer cells <i>in vivo</i>

Additional file 5: of Serum metabolomic profiling predicts synovial gene expression in rheumatoid arthritis

Figure S4. Correlation between serum metabolites and synovial APRIL, CD138, SDF1, IgKappa, and IgMHC. (TIFF 14826 kb