Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Differences between intrinsic and acquired nucleoside analogue resistance in acute myeloid leukaemia cells

doi: 10.1186/s13046-021-02093-4

Figure Lengend Snippet: SAMHD1 determines CNDAC sensitivity of primary AML cells. ( A ) Correlation of SAMHD1 protein levels and CNDAC concentrations that reduce cell viability by 50% (IC 50 s) in bone-marrow-derived leukaemic blasts derived from 24 therapy-naïve AML patients. Cells were co-immunostained for CD33, CD34, CD45 (surface markers) and intracellular SAMHD1 and the mean fluorescence intensity (MFI) was analysed by flow cytometry. ATP assays were performed in three technical replicates to determine the CNDAC IC 50 values. Linear regression analyses were performed using GraphPad Prism. ( B ) CNDAC IC 50 values in bone-marrow-derived leukaemic blasts derived from six therapy-naïve AML patients either treated with VPX virus-like particles (VPX-VLPs, cause SAMHD1 depletion), VPR virus-like particles (VPR-VLPs, negative control), or left untreated. Horizontal lines and error bars indicate means ± SD of three technical replicates. p-values were determined by two-tailed Student’s t-tests (* p < 0.05; ** p < 0.01; *** p < 0.001). ( C ) CNDAC dose-response curves in primary AML cells of one exemplary patient (Patient E) treated with VPX-VLPs, VPR-VLPs or left untreated. IC 50 values represent means ± SD of three technical replicates. ( D ) Representative Western blots indicating SAMHD1 levels in primary AML cells derived from Patient E in response to treatment with VPX-VLPs. ( E ) CNDAC-triphosphate (CNDAC-TP) levels as determined by LC-MS/MS in primary AML cells derived from Patient E in response to treatment with VPX-VLPs

Article Snippet: Staining for surface markers (CD33, CD34, CD45) for AML blasts was applied before fixation with the following fluorochrome-conjugated antibodies: CD33-PE and CD34-FITC, both from Miltenyi Biotech (130–111-019, 130–113-178) and CD45-V450 from BD Pharmingen (Heidelberg, Germany, 642,275), all diluted 1:5 per 1 × 10 7 cells, and goat anti-rabbit Alexa-Fluor-660 from Invitrogen, Life technologies (1:200, A-21073) as secondary antibody for SAMHD1 staining.

Techniques: Derivative Assay, Fluorescence, Flow Cytometry, Virus, Negative Control, Two Tailed Test, Western Blot, Liquid Chromatography with Mass Spectroscopy

Journal: Contrast Media & Molecular Imaging

Article Title: Radiosynthesis and Preclinical Evaluation of 11 C-VA426, a Cyclooxygenase-2 Selective Ligand

doi: 10.1155/2019/5823261

Figure Lengend Snippet: Semipreparative HPLC purification of 11 C-VA426 radiolabelling.

Article Snippet: Acetonitrile extracts were centrifuged and supernatant filtered with a MillexGX® syringe system for HPLC (Gilson Italia Srl, 321 pump, UV/VIS-151) injection.

Techniques: Purification

Journal: Contrast Media & Molecular Imaging

Article Title: Radiosynthesis and Preclinical Evaluation of 11 C-VA426, a Cyclooxygenase-2 Selective Ligand

doi: 10.1155/2019/5823261

Figure Lengend Snippet: Analytical HPLC quality control. (a) UV chromatogram of the reference standard VA426; (b) UV chromatogram of 11 C-VA426; (c) radiochromatogram of 11 C-VA426 showing a radiochemical purity ≥95%.

Article Snippet: Acetonitrile extracts were centrifuged and supernatant filtered with a MillexGX® syringe system for HPLC (Gilson Italia Srl, 321 pump, UV/VIS-151) injection.

Techniques: Control

Journal: Contrast Media & Molecular Imaging

Article Title: Radiosynthesis and Preclinical Evaluation of 11 C-VA426, a Cyclooxygenase-2 Selective Ligand

doi: 10.1155/2019/5823261

Figure Lengend Snippet: Stability study of 11 C-VA426 by HPLC analysis, in the plasma of healthy mice ( n = 3), 10 min p.i.

Article Snippet: Acetonitrile extracts were centrifuged and supernatant filtered with a MillexGX® syringe system for HPLC (Gilson Italia Srl, 321 pump, UV/VIS-151) injection.

Techniques: Clinical Proteomics

Journal: Contrast Media & Molecular Imaging

Article Title: Radiosynthesis and Preclinical Evaluation of 11 C-VA426, a Cyclooxygenase-2 Selective Ligand

doi: 10.1155/2019/5823261

Figure Lengend Snippet: Stability study of 11 C-VA426 by HPLC analysis, in the liver of healthy mice ( n = 3), 10 min p.i.

Article Snippet: Acetonitrile extracts were centrifuged and supernatant filtered with a MillexGX® syringe system for HPLC (Gilson Italia Srl, 321 pump, UV/VIS-151) injection.

Techniques: