List of protocols |
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xiii | |
Abbreviations |
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xix | |
I General methods |
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1 Preparation of liposomes |
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3 | (28) |
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J. Lasch, V. Weissig, and M. Brandl |
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1 Handling and storage of lipids |
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3 | (1) |
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4 | (19) |
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4 | (3) |
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7 | (2) |
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Freeze-dried rehydration vesicles |
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9 | (1) |
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Reverse-phase evaporation |
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10 | (2) |
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12 | (4) |
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High pressure homogenization |
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16 | (7) |
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3 Purification of liposomes |
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23 | (4) |
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24 | (1) |
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Purification of liposomes by centrifugation |
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25 | (2) |
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27 | (4) |
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2 Characterization of liposomes |
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31 | (48) |
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Nicolaas Jan Zuidam, Remco de Vrueh, and Daan J.A. Crommelin |
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31 | (2) |
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2 Chemical analysis of liposomal components |
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33 | (17) |
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Spectrophotometric quantification of phospholipids |
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33 | (2) |
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Thin-layer chromatography of lipids |
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35 | (4) |
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HPLC analysis of (phospho)lipids |
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39 | (2) |
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HPLC analysis of cholesterol and a-tocopherol |
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41 | (1) |
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GLC analysis of fatty acids |
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42 | (3) |
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Chemical analysis of degradation of liposomal phospholipids |
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45 | (5) |
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3 Physical characterization of liposomes |
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50 | (22) |
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Determination of percentage capture |
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50 | (4) |
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Determination of percentage release |
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54 | (10) |
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Determination of entrapped volume |
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64 | (1) |
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64 | (3) |
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Size determination of liposomes |
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67 | (5) |
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Zeta potential measurements based on PCS |
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72 | (1) |
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72 | (5) |
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Bligh and Dyer-extraction |
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72 | (2) |
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Sep-Pak minicolumn extraction |
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74 | (1) |
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Preparation of carboxyfluorescein and calcein solutions |
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75 | (2) |
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77 | (2) |
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3 Physical methods of study: differential scanning calorimetry |
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79 | (26) |
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Kevin M.G. Taylor and Duncan Q.M. Craig |
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79 | (1) |
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79 | (5) |
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Differential scanning calorimetry |
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79 | (3) |
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High sensitivity differential scanning calorimetry |
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82 | (1) |
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Modulated temperature differential scanning calorimetry |
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82 | (2) |
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3 Practical aspects of making DSC measurements |
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84 | (8) |
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84 | (1) |
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85 | (1) |
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85 | (1) |
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86 | (1) |
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Conditions required for liposomal studies |
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87 | (2) |
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Practicalities of HSDSC measurements |
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89 | (1) |
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Practicalities of MTDSC measurements |
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90 | (2) |
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4 Application of DSC and related techniques to the study of liposomes |
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92 | (9) |
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Phase transition behaviour of liposomes |
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92 | (3) |
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Phase transition of multi-component systems |
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95 | (1) |
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Phospholipid-drug interactions |
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96 | (3) |
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DSC studies of liposomes as model membranes |
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99 | (1) |
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MTDSC of freeze-dried liposomes |
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99 | (2) |
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101 | (1) |
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101 | (4) |
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4 Fluorescence methods in liposome research |
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105 | (44) |
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Nejat Düzgünes, Luis A. Bagatolli, Paul Meers, Yu-Kyuong Oh, and Robert M. Straubinger |
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1 Fluorescence assays for liposome fusion |
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105 | (13) |
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The terbium/dipicolinic acid assay for intermixing of aqueous contents |
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105 | (7) |
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The aminonaphthalene trisulfonic acid/p-xylylene bis(pyridinium) bromide (ANTS/DPX) assay for aqueous contents mixing |
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112 | (1) |
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The NBD/rhodamine resonance energy transfer assay for lipid mixing |
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113 | (4) |
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A fluorescence assay for intermixing of inner monolayers |
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117 | (1) |
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2 Fluorescence assays for liposome permeability |
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118 | (1) |
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Release of carboxyfluorescein or calcein |
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118 | (1) |
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Release of the Th /DPA complex |
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118 | (1) |
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119 | (1) |
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3 Fluorescent detection of protein binding to phospholipid membranes |
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119 | (7) |
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Detection via membrane-associated fluorophores |
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120 | (3) |
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Detection with intrinsic protein-associated fluorophores |
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123 | (3) |
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4 Liposomes as pH sensors in the study of cellular interactions |
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126 | (9) |
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The hyranine (HPTS) assay |
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126 | (6) |
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The dual-fluorophore assay |
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132 | (3) |
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5 Two-photon excitation fluorescence microscopy to detect lipid phases in giant liposomes |
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135 | (8) |
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Direct visualization of lipid domain coexistence in bilayers |
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135 | (1) |
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Giant unilamellar liposomes |
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136 | (2) |
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Two-photon excitation microscopy |
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138 | (5) |
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Visualization of gel/fluid phase coexistence in binary phospholipid mixtures |
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143 | (1) |
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143 | (1) |
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143 | (6) |
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5 Stability, storage, and sterilization of liposomes |
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149 | (18) |
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Nicolaas Jan Zuidam, Ewoud van Winden, Remco de Vrueh, and Daan J.A. Crommelin |
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149 | (1) |
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2 Prevention of chemical degradation |
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149 | (5) |
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149 | (3) |
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Hydrolysis of phospholipids |
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152 | (2) |
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3 Prevention of physical changes |
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154 | (1) |
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4 Freeze-drying of liposomes |
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155 | (9) |
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155 | (1) |
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Important factors for freeze-drying of liposomes |
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156 | (5) |
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Stability in the freeze-dried state |
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161 | (1) |
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Example of a freeze-drying protocol |
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162 | (2) |
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5 Preparation of sterile liposome formulations |
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164 | (1) |
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164 | (3) |
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6 Encapsulation of weakly-basic drugs, antisense oligonucleotides, and plasmid DNA within large unilamellar vesicles for drug delivery applications |
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167 | (26) |
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David B. Fenske, Norbert Maurer, and Pieter R. Cullis |
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167 | (2) |
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2 Measurement of phospholipid concentration |
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169 | (2) |
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3 The formation of large unilamellar vesicles by extrusion through polycarbonate filters with defined pore size |
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171 | (2) |
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4 Accumulation of weakly-basic drugs within LUV's in response to transmembrane pH gradients |
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173 | (8) |
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5 Liposomal systems for the encapsulation of genetic drugs: long-circulating vectors for the systemic delivery of genes and antisense oligonucleotides |
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181 | (7) |
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188 | (5) |
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7 Surface modification of liposomes |
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193 | (38) |
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Vladimir P. Torchilin, Volkmar Weissig, Francis J. Martin, Timothy D. Heath, and Roger R.C. New |
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193 | (1) |
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2 Protein and peptide attachment to the liposomal surface |
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194 | (22) |
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Binding of proteins and peptides to liposomes via amino groups |
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194 | (10) |
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Binding of proteins to liposomes via sulfhydryl groups |
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204 | (12) |
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3 Binding of carbohydrates and other small molecules to the liposomal surface |
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216 | (2) |
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4 Attachment of diagnostically significant reporter metal atoms to the liposome surface |
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218 | (9) |
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Conjugation of low molecular weight chelate DTPA with lipid for incorporation into the liposomal membrane |
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219 | (4) |
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Loading liposome-associated chelates with metals |
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223 | (4) |
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227 | (4) |
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8 Long-circulating sterically protected liposomes |
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231 | (36) |
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Alexander L. Klibanov, Vladimir P. Torchilin, and Samuel Zalipsky |
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231 | (1) |
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2 Attachment of sterically protecting polymer (PEG) to liposomes |
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232 | (6) |
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Incorporation of PEG into liposomes |
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232 | (3) |
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Attachment of PEG to previously prepared liposomes |
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235 | (1) |
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Dependence of biodistribution on the size of long-circulating liposomes |
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236 | (1) |
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Tumour accumulation of long-circulating liposomes |
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237 | (1) |
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3 Coupling of various ligands to the distal end of liposome-grafted polymer |
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238 | (23) |
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Ligand attachment to hydrazide-activated PEG |
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243 | (3) |
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Ligand attachment to PDP-activated PEG |
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246 | (3) |
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Ligand attachment via COON-activated PEG |
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249 | (1) |
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Coupling of primary amino group-containing ligands to pNP-PEGPE-containing liposomes |
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250 | (3) |
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Ligand attachment by preparation of ligand-PEG-ligand conjugates and their incorporation into liposomes |
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253 | (8) |
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4 Detachable PEG on the liposome surface |
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261 | (1) |
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5 Biological factors responsible for the clearance of PEGylated liposomes |
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262 | (1) |
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263 | (4) |
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9 Liposomes in biological systems |
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267 | (22) |
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Jan A.A.M. Kamps and Gerrit L. Scherphof |
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267 | (1) |
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2 Interaction of liposomes with cells |
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267 | (12) |
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Possible liposome-cell interaction: adsorption, (receptor-mediated) endocytosis, lipid exchange, fusion |
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267 | (2) |
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269 | (1) |
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Cell lines and primary cell cultures |
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270 | (3) |
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Isolation and culturing of liver cells |
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273 | (6) |
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3 Biodistribution of liposomes |
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279 | (7) |
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Animal models; species differences |
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279 | (1) |
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In vivo liposome-cell interactions |
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280 | (1) |
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281 | (1) |
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Liposome kinetics and tissue distribution |
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282 | (4) |
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286 | (1) |
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286 | (3) |
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10 Cationic liposomes in gene delivery |
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289 | (16) |
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Sean Sullivan, Yan Gong, and Jeffrey Hughes |
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289 | (7) |
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296 | (2) |
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3 Quantification of plasmid DNA concentration |
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298 | (2) |
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4 Calculation of NSP ratio |
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300 | (1) |
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301 | (4) |
II Selected topics and applications |
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11 pH-sensitive liposomes |
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305 | (14) |
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Regine Peschka-Süss and Rolf Schubert |
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305 | (2) |
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2 Mechanisms of pH sensitivity |
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307 | (4) |
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311 | (5) |
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Preparation of pH-sensitive liposomes |
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311 | (3) |
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Characterization of pH-sensitive liposomes |
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314 | (1) |
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Cellular association of pH-sensitive liposomes |
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314 | (2) |
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316 | (3) |
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12 Radiolabelled liposomes for imaging and biodistribution studies |
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319 | (18) |
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Beth A. Goins and William T. Phillips |
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319 | (1) |
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2 Instrumentation for tracking radiolabelled liposomes |
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320 | (3) |
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320 | (2) |
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Positron emission tomographic camera |
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322 | (1) |
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322 | (1) |
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3 Methods for labelling liposomes with gamma-emitting radionuclides |
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323 | (6) |
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323 | (1) |
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Radionuclides available for liposome labelling methods |
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324 | (1) |
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Methods for labelling liposomes using radionuclides |
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325 | (4) |
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4 Characterization of radiolabelled liposomes |
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329 | (1) |
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In vitro radiolabel stability evaluation |
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329 | (1) |
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In vivo radiolabel evaluation |
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329 | (1) |
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5 Image acquisition and analysis methods |
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330 | (4) |
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6 Tissue biodistribution method using gamma well counter |
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334 | (1) |
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335 | (2) |
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13 Isothermic titration calorimetry (ITC) |
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337 | (16) |
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337 | (1) |
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2 Instrument and the principle of technique |
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337 | (3) |
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3 Computer-assisted measurement with the MCS ITC |
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340 | (2) |
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Making a calorimetric titration |
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340 | (2) |
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4 Estimation of critical micellar concentration |
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342 | (5) |
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342 | (2) |
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344 | (3) |
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5 Surfactant-bilayer membrane equilibria |
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347 | (4) |
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Surfactant partitioning into lipid membranes |
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347 | (2) |
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Solubilization of lipsomes |
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349 | (2) |
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351 | (2) |
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14 Vesicular phospholipid gels |
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353 | (20) |
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353 | (2) |
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Biopharmaceutical and technological challenges with liposomes for i.v. application |
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353 | (1) |
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Biopharmaceutical and technological challenges with liposomes for loco-regional administration |
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354 | (1) |
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Phospholipid dispersions: swelling and phases |
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354 | (1) |
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Mechanical stress: formulation of vesicle dispersions and vesicular phospholipid gels |
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354 | (1) |
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The vesicular phospholipid gel concept |
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355 | (1) |
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2 Preparation of vesicular phospholipid gels |
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355 | (3) |
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VPC's made of a single phospholipid |
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355 | (2) |
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VPG's made of two or more lipids |
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357 | (1) |
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3 Loading of VPC's with drugs |
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358 | (3) |
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359 | (1) |
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359 | (1) |
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Incorporation/association |
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360 | (1) |
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361 | (1) |
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5 Characteristics of VPG's |
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361 | (4) |
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361 | (1) |
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362 | (1) |
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Sustained release behaviour in vitro |
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362 | (3) |
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6 Preparation of SUV dispersions from VPG's |
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365 | (1) |
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7 Characterization of SUV dispersions prepared from VPG's |
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365 | (2) |
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Vesicle size distribution |
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365 | (1) |
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366 | (1) |
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367 | (4) |
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Preparation and in vitro characterization of Gemcitabine-containing VPG |
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368 | (1) |
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Intra-tumoral application of Gemcitabine-containing VPG |
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369 | (1) |
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i.v. application of Gemcitabine-containing VPG |
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369 | (2) |
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371 | (2) |
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15 Liposome-based DNA vaccines: procedures for entrapment |
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373 | (8) |
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Gregory Gregoriadis, Brenda McCormack, Yvonne Perrie, Andrew Bacon, Wilson Caparros-Wanderley, and Brahim Zadi |
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373 | (1) |
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374 | (1) |
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3 Entrapment of plasmid DNA into liposomes by the dehydration-rehydration procedure |
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374 | (1) |
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374 | (1) |
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375 | (5) |
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380 | (1) |
Appendix |
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381 | (8) |
Index |
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389 | |