Pharmaceutical Bioassays – Methods and

Shiqi Peng is a Professor and Dean at the School of Chemical Biology and Pharmaceutical Sciences of Capital Medical University in Beijing. He is the author of over 220 journal articles, seven books in China, and over 110 Chinese patents. Ming Zhao is a Professor and Vice Dean at the School of Chemical Biology and Pharmaceutical Sciences of Capital Medical University in Beijing. She is the author of over 120 journal articles, seven books in China, and over 110 Chinese patents.

Preface. Contributors. 1. Methods and Application of Anticancer Bioassays (Shiqi Peng). 1.1 MTT Assay for Six Carcinoma Cells. 1.2 Flow Cytometric Assay for Cell Apoptosis. 1.3 DNA Fragmentation Assay. 1.4 Bcl-XL/BH3 Interaction Assay. 1.5 Dissociation-Enhanced Lanthanide Fluoro-Immunoassay (DELFIA) . 1.6 Ishikawa Cell and Rat Assay for Detecting Antiestrogens. 1.7 ATP Assay for Eight Cells. 1.8 APh Activity Assay. 1.9 Tumor Endothelial Cell Tube Formation Assay. 1.10 Anti-Angiogenic Assay. 1.11 In Vivo Hollow Fiber Assay. 1.12 VX2 Rabbit Lung Assay. 1.13 Insulin-Like Growth Factor-I Induced Kinase Receptor Activation Assay. 1.14 Insulin-Like gD*trkA-Induced Kinase Receptor Activation Assay. 1.15 UV Spectra-Based Calf Thymus DNA Intercalation Assay. 1.16 Fluorescence Spectra-Based Calf Thymus DNA Intercalation Assay. 1.17 P-Glycoprotein Pump in MCF-7R Cells Assay. 1.18 P-Glycoprotein Pump-Related Efflux Carriers Assay. 1.19 [ 3 H]Substrate Transport Inhibition Assay. 1.20 Lactate Dehydrogenase Release Assay. 1.21 Functional Assay of Mitochondrial P-gp . 1.22 Resistance Index Value Assay. References and Notes. 2. Methods and Application Antiviral Assay (Chunying Cui, Shiqi Peng and Ming Zhao). 2.1 Nonradioactive HIV-1 RT Activity Assay. 2.2 Respiratory Syncytial Virus Assay. 2.3 Influenza Virus Types A and B Assay. 2.4 Nasal Exhaled NO Concentration Assay. 2.5 Nasal NOS2 mRNA Quantity Assay. 2.6 RT-PCR and Swine Assay for Anti-HEV Antibody. 2.7 HIV-1 Protease and Reverse Transcriptase Kinetic Assay. 2.8 Anti-HIV Assay. 2.9 Robust Antiviral Assays . 2.10 HIV/SIV Fusion Assay. 2.11 Rapid DNA Hybridization Assay. 2.12 Antiviral Screening Assay for HepAD38 Cell Cultures. 2.13 Trak-CTM HCV Core Assay . References and Notes. 3. Methods and Application of Antitubercular Assay (Chunying Cui, Shiqi Peng and Ming Zhao). 3.1 Mycobacterium tuberculosis Assay. 3.2 DNA Polymerase ss Lyase Assay. 3.3 Agar Dilution Assay for In Vitro Antitubercular Activity. 3.4 Microplate Alamar Blue Assay for In Vitro Antitubercular Activity. 3.5 Radiometric Respiratory Assay for In Vitro Antitubercular Activity. 3.6 Mycobacterium bovis BCG Inhibition Assay . Reference and Notes. 4. Methods and Application of Thrombus Related Assays (Ming Zhao). 4.1 In Vitro Anti-Platelet Aggregation Assay. 4.2 Fibrinolytic Area Assay. 4.3 TXB 2 and PGD 2 TLC Assay. 4.4 TXA 2 Synthase Activity Assay. 4.5 [Ca 2+ ]i Measuring Assay. 4.6 Arachidonic Acid Liberation Assay. 4.7 Serotonin Secretion Assay. 4.8 cAMP Release Assay. 4.9 Ex Vivo Anti-Platelet Aggregation Assay for Patients. 4.10 ATP Release Assay. 4.11 PAF-Induced Mice Mortality Assay. 4.12 PGE 2 and TXB 2 ELISA. 4.13 Thrombelastograph Assay. 4.14 Image-Monitored FeCl 3 -Induced Thrombosis Assay for Rat. 4.15 Weight-Monitored FeCl 3 -Induced Thrombosis Assay for Rats. 4.16 Occlusion Time-Monitored FeCl 3 -Induced Thrombosis Assay for Pig. 4.17 Doppler Blood Flow-Monitored FeCl 3 -Induced Thrombosis Assay for Mouse . 4.18 Rat Groin Flap Assay. 4.19 Ferret Acute Thrombosis Assay. 4.20 Rat Acute Thrombosis Assay. 4.21 Arteriovermus Shunt Assay. 4.22 Plasma Clotting Time Assay. 4.23 Thromboembolic Photochemical Assay for Repeated Stroke in Mice. 4.24 Euglobulin Clot Lysis Assay. 4.25 Clot Formation and Lysis (CloFAL) Assay. 4.26 Fibrin Microplate Assay. 4.27 Fibrinolytic Activity Assay. 4.28 Fibrinolysis Assay. 4.29 Thrombolytic Assay. References and Notes. 5. Methods and Application of Anticoagulation Assays (Ming Zhao). 5.1 Ecarin Chromogenic Assay. 5.2 Anticoagulation Activity Assay in An In Vitro System. 5.3 Anticoagulation Activity Assays in An In Vivo System . 5.4 Rat Thrombosis Assay. 5.5 In Vivo Microvascular IVC Blood Flow Assay. 5.6 Owren PT Assay. 5.7 Rabbit Double-Balloon Injury Assay. 5.8 A Rapid Point-of-Care Assay for Enoxaparin. 5.9 Thromboplastin Clotting Assay. 5.10 Rat Assay for Reproducible Stasis-Induced Venous Thrombosis. 5.11 In Vivo ACT II/Ecarin Clotting Time Assay. 5.12 Ex Vivo and In Vivo Anticoagulation Assay. 5.13 Plasma-Based Ecarin Clotting Time Assay for r-Hirudin . 5.14 Protamine Titration for Heparin in Whole Blood. 5.15 Automated Assay Evaluating Response of Kaolin ACT to Heparin. 5.16 Platelet/Monocyte Interaction-Based PM Exposure Mouse Assay. 5.17 Mouse Tail-Bleeding Time Assay. 5.18 MRI Assay for Rabbit Atherosclerotic Lesions. 5.19 Spiral Computed Tomography Assay. 5.20 Duplex Ultrasound Assay. 5.21 Standard Hemochron Assay. 5.22 Platelet Serotonin Release Assay. 5.23 Activated Partial Thromboplastin Time Assay. 5.24 Rat Stroke Outcome Assay. References and Notes. 6. Methods and Application of Blood Pressure Related Assay (Ming Zhao). 6.1 Human Plasma New Pressor Protein Assay. 6.2 Pulmonary Hypertension Assay. 6.3 Coronary Arteries Constriction Assay. 6.4 MRI and Brain Natriuretic Peptide Assays . 6.5 Competition Enzyme-Linked Immunosorbent Assay. 6.6 Right Ventricular Pressure Assay. 6.7 Plasma Nitrite/Nitrate Concentration Assay. 6.8 Adeno-Associated Virus Vector-Caused Rat Pulmonary Artery Pressure Assay. 6.9 Adeno-Associated Virus Vector-Caused Rat Protein and mRNA Assay. 6.10 N-Terminal pro-Brain Natriuretic Peptide (N-T proBNP) Assay. 6.11 Middle Cerebral Artery Occlusion Assay. 6.12 Vascular Endothelial Growth Factor Level Assay. 6.13 Antispasmodic Agent In Vivo Action Assay. 6.14 Temperature Assay in Awake Subjects. References and Notes. 7. Methods and Application of Assays Related to Parkinson's Disease and Graves' Disease (Shiqi Peng). 7.1 Flow Cytometric Assay for Cellular DNA Content and Caspase-3. 7.2 Swine Resuscitation Assay. 7.3 HTLV-tax1 or pMuLV-SV-nlslacZ Vectors Transfected Cell Assay. 7.4 Parkinsonian Rat Assay. 7.5 ELISA for Nerve Growth Factor Antigen. 7.6 Assay for ss-Nerve Growth Factor Levels in Cerebrospinai Fluid. 7.7 MDCK Scatter Assay. 7.8 Facial Nerve and Spinal Root Avulsions. 7.9 TRAb Assays. 7.10 Human Thyrotropin Receptor (hTSHR) Assay. 7.11 Soluble ICAM-1, TSAb and TBIAb Activity Assays. 7.12 Microarray Immunoassay for hTSHr Production. 7.13 Affinity Assay for [ 35 S]GTPgammaS Binding to Gas/olf. 7.14 Tissue Segment Binding Assay for alpha1B-Adrenoceptor. 7.15 Membrane Binding Assay with Rat Cerebral Cortex for alpha1B-Adrenoceptor . 7.16 Whole Cell Binding Assay for alpha1B-Adrenoceptor. 7.17 Functional Assay for alpha1A,B-Adrenoceptor. 7.18 Rat Neuroprotective Assay. 7.19 GABA-Benzodiazepine Receptor Assay. 7.20 Forced Swimming and Tail Suspension Assays. 7.21 IGF-I Kinase Receptor Activation (KIRA) Assay. 7.22 gD.trkA Kinase Receptor Activation (KIRA) Assay. 7.23 gD.trk KIRA-ELISA. 7.24 Assays for Cannabinoid Receptors in Rat Cerebella or Mouse Brains . References and Notes. 8. Methods and Application of Alzheimer's Disease Assay (Shiqi Peng). 8.1 Assay for Oxidative Stress in Cerebral Cortex of AD Mice. 8.2 Reporter Assay for Primary Neuronal Cultures . 8.3 Electrophoretic Mobility Shift Assay (EMSA). 8.4 Binding Assays Using Aggregated Ass peptide in Solution. 8.5 Assay for Muscarinic Receptor 1 in Alzheimer's Dementia Model . 8.6 ss-Secretase Activity Assay. 8.7 Ass Fibril Binding Assay. 8.8 TLC and Microplate Assays for Acetylcholinesterase Inhibitors. 8.9 Immunocapture Assay Measuring Specific Enzyme Activity of Neprilysin. 8.10 In Vivo AChE Inhibition Assay. 8.11 Single Particle Assay for Ass Aggregates. 8.12 Indirect Immunofluorescence Assay. 8.13 Mouse Behavioral Assays. 8.14 Center of Pressure (CoP) Assay in Mice. 8.15 Assay for Plasma Levels of DJ-1. 8.16 Membrane Filter Assay for Tau Aggregation. 8.17 Assays for Motor Neuron Degeneration. 8.18 HPLC Assay for Neuroprotective Agent in Mice Plasma. 8.19 Tissue Culture Assays for SOD1 Mutations. 8.20 Luciferase-Based Reporter Assay. References and Notes. 9. Methods and Application of Antiosteoporosis Assay (Ming Zhao). 9.1 Rat Bone Mineral Density Assay. 9.2 Osteoblastic Cell Proliferation and Alkaline Phosphatase (ALP) Activity Assays. 9.3 Murine Osteoblastic MC3T3-E1 Cell Calcification and Van Kossa Assays. 9.4 Osteoclast Generation Assay for Male Senile Rat. 9.5 Bone Resorption and Recovery Related Assays. 9.6 Mouse Bone Mineral Density Assay. 9.7 PPAR-gamma Competitor Assay. 9.8 Human Serum Estrogen Level Assay. 9.9 Luciferase Activity Assay. 9.10 IL-1ss and TNF-alpha Level Assay. 9.11 ER Binding and Receptor Activity Assays. 9.12 Fluorescent Estrogen Receptor Assay. 9.13 ELISA for Urinary Helical Peptide. 9.14 Urine Midmolecule Osteocalcin Assay. 9.15 BMD and Osteocalcin Assay. References and Notes. 10. Methods and Application of Immunomodulating Assay (Shiqi Peng). 10.1 Rat Mast Cell Histamine-Release Assay. 10.2 Rabbit Aortic Force Assay. 10.3 Dopaminergic Cell Death-Based Neural Transplantation Assay. 10.4 Mast Cell Degranulation Assay. 10.5 Basophils Assay as Allergen. 10.6 RBL-2H3 Cell Desensitization Assay. 10.7 Migration Assay of Dendritic Cell from PBMCs. 10.8 Mouse EAE Induction Assay. 10.9 COSTIM Assay for DC/T-Cells. 10.10 Cytokine Assay for L-6, IL-10, IL-12, and TNF-alpha of DCs. 10.11 ELISPOT Assay for DC IFN-gamma . 10.12 DC Function Assay for Evaluating Toll-like Receptor Function . 10.13 Migration Assay of Dendritic Cell from Bone Marrow of A/J Mice . 10.14 Lymphoid Organ Assay. 10.15 ELISA of IFN-gamma from Human Myelomonocytic KG-1 Cells. 10.16 Human Whole Blood IFN- gamma Assays . 10.17 Sheep Whole Blood IFN- gamma Assays. 10.18 ELISPOT Assay for IFN- gamma. 10.19 IFN-ss RG Assay. 10.20 Anti-rHuEPO NAb Assay. 10.21 Chloramphenicol Acetyltransferase Assay. 10.22 Chemotaxis Assay . 10.23 Fibroblast-Populated Microsphere Assay. 10.24 Fibroblast-Populated Concentric Microsphere Assay. 10.25 Radial Assay of Chemotaxis . 10.26 Antibody Forming Cell Assay. 10.27 Immunosuppressive Assay. 10.28 Cell-Based ELISA. 10.29 Large Animal Lung Transplantation Assay. References and Notes. 11. Methods and Application of Anti-inflammatory Assays (Ming Zhao). 11.1 Adhesion Formation Assay. 11.2 Ligand Complex-Based Adhesion Assay. 11.3 Human Umbilical Vein Endothelial Cell Assay. 11.4 Pleurisy Mouse Assay. 11.5 Proliferation of PBMC Assay. 11.6 COX-1, COX-2 and 5-LOX Assay with [1- 14 C]Arachidonic Acid. 11.7 COX-1 and COX-2 Assay with Human Whole Blood. 11.8 o-Hydroxyleukotriene B4 Assay. 11.9 Leukocyte Rolling and Adherent of Leukocytes Assay. 11.10 CCR5 Receptor Binding Assay. 11.11 Tissue Binding Affinity Assay. 11.12 G93A-SOD1 Transgenic Mouse Assay. 11.13 MCP-1 Induced ERK1 and ERK2 Phosphorylation Assay. 11.14 LPS and IL-6 Induced ERK1 and ERK2 Phosphorylation Assay. 11.15 ELA4.NOB-1/CTLL Cell Assay . 11.16 FK506 Binding Protein 51 (FKBP51) mRNA Assay. 1.17 Xylene-Induced Ear Edema Assay. References and Notes. 12. Methods and Application of Antioxidant Activity Assay (Shiqi Peng). 12.1 Blood and Plasma Total Antioxidant Capacity (TAC) Assay. 12.2 Ferric Reducing-Antioxidant Power (FRAP) Assay. 12.3 Human LDL Oxidation Assay. 12.4 DPPH Radical Cation Scavenging Assay. 12.5 ABTS+ Radical Cation Scavenging Assay. 12.6 Lipid Peroxidation Assay Using Rat Brain Tissue. 12.7 Flow-Through Chemiluminescence (FTCL) Assay. 12.8 Superoxide Radical Scavenging Assay. 12.9 Deoxyribose Assay for Hydroxyl Radical Scavenging Activity. 12.10 DNA Nicking Assay for Hydroxyl Radical Scavenging Activity. 12.11 Oxidative Lag-Time Assay. 12.12 TBARS and Electrophoresis Assay. 12.13 Reporter and Electrophoretic Mobility Shift Assay (EMSA). 12.14 [Ca 2+ ]cyt Assay. 12.15 Quantitative Real-Time PCR Assay. 12.16 FT-IR Based Assay for Antioxidation Activity of Ionol and Piperidone. 12.17 HPLC Assay for Antioxidation Potential of Polyphenol. 12.18 ROS Production Assay. 12.19 Rabbit LDL Oxidation Assay. 12.20 ROS Scaverging Assays. 12.21 DNA Damage Assay. 12.22 Egg Yolk TBARS Assay. 12.23 Mouse Catalase (CAT) Assay. 12.24 Rat Tissue TBARS Assay. 12.25 Antioxidant Activity Assay for ss-Carotene/Linoleic Acid System. 12.26 Rat Brain Tissue NO Assay. 12.27 Rat Brain Antioxidative Enzyme Assay. 12.28 Rat Brain Hippocampi Protein Oxidation Assay. References and Notes. 13. Methods and Application of Analgesic Assays (Ming Zhao). 13.1 Algogenic Activity Assay for Rat Ureteral Stone. 13.2 Cold Pressor-Based Assay for Acute Pain of Healthy Volunteers. 13.3 Heat-Based Assay for Acute Pain of Healthy Volunteers. 13.4 Electrical Stimulation-Based Assay for Acute Pain of Healthy Volunteers. 13.5 Radiant Heat Tail-Flick Assay for Mice. 13.6 Pain Behaviors/Responses Assays in Rats. 13.7 Hot Plate Assay in Rats. 13.8 Plantar Assay in Rats. 13.9 Hot Plate Assay in Mice . 13.10 Paw and Tail Formalin Assays in Mice. 13.11 Rat Assays for Bone Cancer Pain . 13.12 Mouse Assay for Hind Paw Cancer Pain . 13.13 Visceral Pain Assay. 13.14 Canine Nociceptive Thermal Escape Assay in Dog. 13.15 Carrageenan Assay in Rats. 13.16 Electrophysiological Assay for Mice with Tumor-Evoked Hyperalgesia. 13.17 Mouse Assay for Bone Cancer Pain. References and Notes. 14. Methods and Application of Epilepsy Assays (Ming Zhao, Shiqi Peng and Guohui Cui, ). 14.1 HISS Assay. 14.2 Mouse Locomotor Activity Assay. 14.3 Mouse Diathesis-Stress Assay. 14.4 Social Interaction Assay. 14.5 Spatial Learning Ability Assay for Recurrent Seizure Rats. 14.6 Timed PTZ Infusion Assay for Mice. 14.7 Timed PTZ Seizure Assay for Mice. 14.8 Maximal Electroconvulsions Threshold Assay for Mice. 14.9 MES Assay. 14.10 6-Hz Psychomotor Seizure Assay for Mice. 14.11 Subcutaneous Bicuculline and Picrotoxin Assay for Mice. 14.12 NMDA-Induced Convulsions Assay for Mice. 14.13 AGS Assay. 14.14 Kindled Rat Assay for Focal Seizures. 14.15 Cobalt/Homocysteine Assay for Status Epilepticus of Rats. 14.16 PTZ-Induced Kindling Assay. 14.17 ICES Assay for Mice. References and Notes. 15. Methods and Application of Diabetes Assay (Ming Zhao, Shiqi Peng and Guohui Cui). 15.1 Islet Xenograft Assay for Diabetic Mice. 15.2 Assays for Spontaneous Diabetes and Adoptive Transfer of Diabetes. 15.3 Oral Glucose Tolerance Assays for Patients. 15.4 Injection Glucose Tolerance Assays for Rats. 15.5 Renal Cortical TGF-ss1 Protein Assays for Rats. 15.6 Low-Dose Streptozotocin Treated Heminephre Ctomized Rat Assay. 15.7 Rat Early Diabetic Nephropathy Assay. 15.8 Urinary Endothelin-1 Excretion Assay for Type 2 Diabetes Rats. 15.9 Subtotally Nephrectomized Rat Assay. 15.10 Type 2 Diabetes Mice Assay. References. 16. Methods and Application of Assay for Toxins from Microorganism (Shiqi Peng). 16.1 Colorimetric Yeast Assay for Trichothecene Mycotoxins. 16.2 Colorimetric Cell Proliferation Assay. 16.3 Yeast DEL Assay. 16.4 NCCLS and EUCAST Assays . 16.5 Alcohol Dehydrogenase Based Colorimetric Assays. 16.6 Colorimetric Assay for Iron in Yeast. 16.7 Mcroplate Redox Assays of E. Coli. 16.8 Ciliate Tetrahymena Thermophila Assay for Trichothecene Mycotoxins. 16.9 Fluorescent Dyes-Based Cell Viability Assay for Triton X-100 Toxicity. 16.10 MTT Assay for Fusarium Mycotoxins. 16.11 Mortality and Frass Production Assay for Toxicity of Bacterial Strains . 16.12 Cell Toxicity Assay. 16.13 Dhase Inhibition Assay. 16.14 Sediment Toxicity Assay . 16.15 Toxicity Assay of Particle-Associated Arsenite and Mercury. 16.16 Genetic Toxicity Assay. 16.17 Microtox Assays. 16.18 DNA Piezoelectric Biosensor Assay. 16.19 Protein Phosphatase Inhibition Assay and ELISA of Microcystins. 16.20 Lepidium Sativum Assay for Microcystin Toxicity. 16.21 Antiproliferative Assay for Interleukin-4. References and Notes. 17. Methods and Application of Toxicity Assay for Chemicals (Shiqi Peng). 17.1 Lux-Fluoro Assay for Combined Genotoxicity and Cytotoxicity of Chemicals. 17.2 Flow Cytometry and Microscopy Based Assay. 17.3 Cell Transformation Assay. 17.4 ALIC-Based Assay for Toxicity of Chemicals in SPM. 17.5 Immunization in the Murine Assay. 17.6 Immunotoxicological Functional Assay. 17.7 PWM-Induced IgM Assay for Toxicity of Chemicals. 17.8 Tetrachlorodibenzo-p-dioxin (TCDD)-Induced Toxicity Assay. 17.9 H4IIE EROD Assay. 17.10 Rat Mutation Assay. 17.11 AhR and Related Assay. 17.12 Ectonucleotidase Expression Assay. 17.13 Toxicity Equivalent Quantity (TEQ) Assay. 17.14 Enzyme Lacking AA Epoxygenase Activity Assay. 17.15 Japanese Medaka Embryo-Larval Assay. 17.16 Green Fluorescent Protein-Based Cell Assay. 17.17 RACB Assay for Ovarian Toxicity of Xenobiotics. 17.18 Uroepithelial Cell Assay. 17.19 Male Rat Systemic Toxicity Assay. 17.20 28-Day Oral Toxicity Assay. 17.21 Endocrine Disruptors Assay. 17.22 Mating Efficiency Assay. 17.23 Soils Assay for Contaminants and Toxicity. 17.24 Rice Nitrate Reductase Activity Assay. 17.25 Tradescantia-Micronucleus Assay. 17.26 Nitric Oxide Neurotoxicity Assay. 17.27 EBV-Transformed Human Burkitt's Lymphoma Cell Assay. 17.28 Murine Bone Marrow Assay for Haemopoietic and Osteogenic Toxicity. 17.29 PNAR-GFP Assay for Carcinogenic Toxicity of Nitrate. 17.30 Brine Shrimp (Artemia Salina) Assay. 17.31 DPPH Radical Scavenging Property Assay. References and Notes. 18 Methods and Application of Hepatoxicity and Hepatoprotective Assay (Shiqi Peng). 18.1 GST-P Enzyme-Altered Foci Assay. 18.2 GST-P+ and TGF-alpha+ Foci Assay. 18.3 Partial Hepatectomy Assay. 18.4 In Vivo Short-Term Liver Initiation Assay in Rats. 18.5 TUNEL Related Aassay. 18.6 Haematopoietic Progenitor Cells Mobilization Assay. 18.7 Electrophoretic Mobility Shift Assay. 18.8 Telomerase Activity Assay . 18.9 Von Willebrand Factor (vWf) Level Assay. 18.10 Bromodeoxyuridine Incorporation Assay. 18.11 NF-kB Activation Assay . 18.12 Rat Summation of Initiation Activity Assay. 18.13 Hepatoprotective Assay. 18.14 Cytokine Gene Expression Assay. 18.15 Competitive Inhibition Assay for Immunodominance of O-Specific Polysaccharides of Gram-Negative Bacilli. 18.16 Marine Toxin Okadaic Acid (OA) Assay and ELISA. 18.17 Anti-GST-CTX MVIIA Antiboy in Omega-Conotoxin MVIIA Assay. 18.18 Assays for Rituximab in Patient Plasmas. 18.19 Assays of TNF-alpha, Superoxide and Thymocyte for Liver Injury of Mice. 18.20 Gene Transfer of Kringle 1-5 and Related Assays for Mice with Hepatocellular Carcinoma. 18.21 Body Weight Assay for Mice with Liver Tumor Incidence. 18.22 P450 Activity and Inducibility in Rat Hepatocytes for Cytotoxicity Assay. 18.23 LDH Release in Rat Hepatocytes for Toxicity Assay. 18.24 Rainbow Trout Hepatocytes' EROD Activity Assay. 18.25 EROD Activity, ROS Production and Cytotoxic Concentration in Fish Hepatocytes for Drug Toxicity Assays. 18.26 Coho Salmon's IGF-I Gene Sequencing and TaqMan Assay. 18.27 DNA Strand Break in Rat Hepatocyte and Comet Assay. 18.28 DNA Laddering in Rat Hepatocyte and TUNEL Assay. 18.29 Counting Increases in Viable Cell Number for Hepatocyte Proliferation Assay. 18.30 Assay of HGF and TGF-b in CCL-64 Cells. 18.31 Assay for Serum IFN-a of Patients with Chronic Hepatitis C. 18.32 Rat Medium-Term Liver, DNA Microarray and Cup-Reducing Antioxidation Assays for DEN-Induced Hepatocarcinogenesis. 18.33 Rat Medium-Term Liver Assay for DEN-Induced Hepatocarcinogenesis Assay. 18.34 Assay for Rat Carcinogenesis Initiated by Three Carcinogens. 18.35 Assay for Rat Carcinogenesis Initiated by Five Carcinogens. 18.36 Assay for DEN-Initiated Rat Carcinogenesis. 18.37 Assay for Normal Dose DEN-Initiated and Low Dose DEN-Maintained Rat Carcinogenesis. 18.38 TGF-b-Mediated Antiproliferation Assay. 18.39 Assay for DEN and HCB-Initiated Rat Carcinogenesis. References and Notes. 19 Methods and Applications of Estrogen Assays ( Shiqi Peng). 19.1 Yeast Estrogen Assay. 19.2 In Vivo, Ex Vivo, and In Vitro Assays for Estrogen-Like Effect. 19.3 Ameliorative Yeast Assay. 19.4 In Yeast Two-Hybrid Assay. 19.5 Estrogen-Sensitive Yeast Strain RMY/ER-ERE Assay. 19.6 Bioluminescent Yeast Assays. 19.7 LYES Assay. 19.8 ERa and ERb Stable Transactivation Assay. 19.9 HELNa and HELNb Transfected Cell Assay. 19.10 Luciferase Reporter Gene Assay. 19.11 Target AKT Pathway Assay. 19.12 Luciferase Assay. 19.13 Androgen Receptor Transactivation Assay. 19.14 Green Fluorescent Protein Expression Assay. 19.15 Luteinizing Hormone Releasing Hormone (LHRH) Assay. 19.16 Ishikawa Endometrial Cancer Cell Assay for Estrogen Receptor Expression. 19.17 Ishikawa Cell Assay for 11b-HSD2 Activity. 19.18 Ishikawa Cell Assay for Estrogen Activity. 19.19 Ishikawa Cell Assay for Plasminogen Activator Inhibitor-1 (PAI-1). 19.20 Testosterone Induction Assay with Sertoli Cell. 19.21 Electrophoretic Mobility and Antibody Shift Assays. References and Notes. 20 Methods and Applications of Antimalarial Assays ( Shiqi Peng). 20.1 Plasmodium falciparum and Murine P388 Leukemia Cell Assay. 20.2 Antiplasmodial Activity Assay. 20.3 Plasmodium falciparum Growth In Vitro Assay. 20.4 Antibody Assay for Red Blood Cell Polymorphisms. 20.5 Leishmania Macrophage Assay. 20.6 Lactate Dehydrogenase-Based Antiplasmodial Assay. 20.7 Histidine-Rich Protein II Assay. 20.8 Histidine-Rich Protein II Assay. 20.9 Antimalarial Activity Assay. 20.10 Plasmodium yoelii Liver Stage Parasites Inhibition Assay. 20.11 In Vivo Antimalarial Assay. 20.12 b-Hematin Inhibition Assay. 20.13 Non-radiolabeled Ferriprotoporphyrin IX Biomineralization Inhibition Assay. 20.14 Survival of Anopheles gambiae Assay. 20.15 Chloroquine Assay. 20.16 MS Assay for Quantification of Chloroquine in Dog Plasma. 20.17 Sensitive Fluorescence HPLC Assay for AQ-13. 20.18 HPLC and HPTLC Assays for Chloroquine, Primaquine, and Bulaquine. 20.19 Real-Time PCR-Based Chloroquine Sensitivity Assay. 20.20 Rat Embryos In Vitro Assay. 20.21 HPLC-MS Assay for b-DHA in Rat Plasma. 20.22 Plasmodium falciparum Clone and DHA Assay. References. 21 Methods and Applications of Cytogenetic Receptor and Enzyme Assays ( Shiqi Peng). 21.1 UT-7/EPO Cell Proliferation and Neutralizing Anti-EPO Antibody Assays. 21.2 Assay System for Biotin Protein Ligase (BPL) from Escherichia coli. 21.3 b-Galactosidase and a-Amylase Biosynthesis Assays. 21.4 Penicillin-Binding Protein (PBP 5) and Vancomycin Activity Assays. 21.5 Recombinant Bacteria Assay Evaluating Androgen Biosynthesis. 21.6 UDP-Glc 4-Epimerase Assay. 21.7 ATPs Exchange Assays. 21.8 Lux-Fluoro Assay for Combined Genotoxicity and Cytotoxicity. 21.9 Reporter Gene Assay for Aryl Hydrocarbon Receptor. 21.10 CYP1A Enzyme Assay. 21.11 CYP1A Activity (EROD) Assay. 21.12 Deubiquitinating Enzyme (DUB) Activity Assay. 21.13 Flow Cytometric Assay. 21.14 Serum Neutralization Assay Based on rHMPV-GFP. 21.15 Yellow Fluorescent Protein-Based Assay. 21.16 Red Fluorescent Cytotoxic T Lymphocyte Assay. 21.17 Green Fluorescent Protein-Based Assay. 21.18 DNA Break Assay in HepG2 Cells. References and Notes.