3.5.4.1 additional information - capability of human adipose tissue-derived mesenchymal stem cells (AT-MSC) as vehicles for gene-directed enzyme prodrug therapy, fusion product of cytosine deaminase derived from yeast in combination with systemic 5-fluorocytosine (5-Fc), expression and antitumoral effects of fusion constructs tested in cell lines and xenograft models, in vivo analysis in immunodeficient mouse model, regression in 89% of tumors achieved, therapeutic cell homing into subcutaneous melanoma and inhibition of tumor growth 699300 3.5.4.1 additional information - cytosine deaminase of Escherichia coli used as a negative selection marker in Streptomyces, can be applied for any actinobacterial species naturally resistant to 5-fluorocytosine while sensitive to 5-fluorouracil, valuable addition to the actinobacterial genetic toolbox 695762 3.5.4.1 additional information - development of cell lines and animal xenograft models, expression and function of fusion constructs between cytosine deaminase and monomeric DsRed (mDsRed), analyzed by Western blot, flow cytometry, fluorescence microscopy, magnetic resonance spectroscopy, analysis of antitumoral effect of 5-fluorouracil (5-FU) and 5-fluorocytosine (5-FC) in transgenic background, increase of cytosine deaminase enhances the potential of gene-directed enzyme prodrug therapy, conversion of 5-fluorocytosine to 5-fluorouracil analyzed in mice bearing tumor xenografts with the fusion product between cytosine deaminase and monomeric DsRed (R3327-CD/mDsRed) 684230 3.5.4.1 additional information - incubation of transfected cells increasing concentrations of free or liposomal 5-fluorocytosine (5-FC) for 72 h, effectivness gene-directed enzyme prodrug therapy (GDEPT) tested in mice transplantated with MC-38 tumor cells, survival times of mice bearing tumors, local concentration and toxicity of liposomal prodrug in different organs, enhancement of antitumor effects by liposomal form of 5-fluorocytosine (5-FC) 685930 3.5.4.1 additional information - pH-indicator assay for the measurement of the steady-state kinetic parameters of deamination reaction 667664 3.5.4.1 additional information - selection and characterization of a human monoclonal antibody in single chain fragment (scFv) format for detection of expression of cytosine deaminase in GDEPT/ADEPT studies, selection and characterization of specific single chain fragment variable (scFv) antibody specific for cytosine deaminase, cytotoxic assay, amino acid sequence of the CDR3 regions of the selected antibody and genetic structure of phage antibody from ETH-2 library determined, human antibody generated by this method to detect cytosine deaminase in routinary laboratory techniques without interfering with its enzymatic function 696961 3.5.4.1 0.1 - - 209488 3.5.4.1 0.12 - (NH4)2SO4 as nitrogen source 209500 3.5.4.1 0.14 - - 209488 3.5.4.1 0.19 - - 209488 3.5.4.1 0.22 - - 209488 3.5.4.1 0.23 - - 209488 3.5.4.1 0.26 - strain K-1 AKU 0011 209488 3.5.4.1 0.31 - - 209488 3.5.4.1 0.32 - strain AKU 0009 209488 3.5.4.1 0.41 - - 209488 3.5.4.1 0.44 - - 209488 3.5.4.1 0.52 - - 209488 3.5.4.1 0.58 - - 209488 3.5.4.1 0.6 - - 209488 3.5.4.1 0.62 - - 209488 3.5.4.1 0.69 - - 209496 3.5.4.1 1 - asparagine as nitrogen source 209500 3.5.4.1 1.2 - - 209487 3.5.4.1 2 - + 0.025 mM Cu(C2H3O2) 209503 3.5.4.1 2.3 - - 209501 3.5.4.1 2.6 - + 0.025 mM HgCl2 209503 3.5.4.1 2.6 - + 0.025 mM Pb(C2H3O2) 209503 3.5.4.1 2.7 - + 0.025 mM CrCl2 209503 3.5.4.1 2.7 - cytosine as nitrogen source 209500 3.5.4.1 2.9 - apoCDase, + 0.025 SnCl2 209503 3.5.4.1 3.1 - - 209484 3.5.4.1 3.1 - + 0.025 mM Cu(C2H3O2) 209503 3.5.4.1 3.2 - + 0.025 mM CaCl2 209503 3.5.4.1 4 - + 0.025 mM NiCl2, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 5.6 - + 0.025 mM NiCl2 209503 3.5.4.1 8.3 - + 0.025 mM Pb(C2H3O2), additional incubation of 0.025 mM Fe2+ 209503 3.5.4.1 9.8 - - 209497 3.5.4.1 11 - + 0.025 mM HgCl2, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 12 - + 0.025 mM Zn(C2H3O2), additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 13 - + 0.025 mM Zn(C2H3O2) 209503 3.5.4.1 17 - - 209493 3.5.4.1 21 - + 0.025 mM CoSO4, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 21.4 - - 209494 3.5.4.1 26 - + 0.025 mM CoSO4 209503 3.5.4.1 27 - - 209503 3.5.4.1 45 - + 0.025 mM MnSO4 209503 3.5.4.1 58 - wild-type enzyme, substrate 5-fluorocytosine 688280 3.5.4.1 62 - + 0.025 mM MnSO4, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 65 - + 0.025 mM FeSO4, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 87 - + 0.025 mM CrCl2, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 87 - + 0.025 mM FeSO4 209503 3.5.4.1 87 - apoCDase, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 88 - + 0.025 mM CaCl2, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 94 - + 0.025 SnCl2, additional incubation with 0.025 mM Fe2+ 209503 3.5.4.1 144 - wild-type enzyme, substrate cytosine 688280