Description
SPRINGER Bioassays In Experimental And Preclinical Pharmacology (Hb 2021) by ARUNACHALAM K.
This detailed book explores protocols for a wide array of preclinical pharmacology and toxicology evaluations to be applied to chemical drugs and their development through in vitro, involving tissues and cell lines, and in vivo models, using animals as experimental systems, utilized to conduct pharmacological research. Written for the Springer Protocols Handbooks series, the methodologies included in this collection have been standardized by the authors through extensive use in the lab so that they are ready to be applied in the labs of readers around the world. Authoritative and practical, Bioassays in Experimental and Preclinical Pharmacology aims to assist undergraduate and postgraduate students, research scholars, scientists, and other academicians performing research in the vital field of drug discovery. 1. Overview2. Cell Culture Assays2.1 Cell Cycle Assay: Propidium Iodide Marking2.2 Apoptosis Assay: Annexin V/Propidium Iodide Marking2.3 Cytotoxicity in Caco-2 cells by the Alamar blue method2.4 Viability in Caco-2 cells stimulated with lipopolysaccharide (LPS)2.5 Determination of Nitric oxide (NO) by in vitro experiment2.6 Analysis of specific cytotoxicity against RAW 264.7 for determination anti-inflammatory activity in vitro2.7 Scratching of Fibroblast cells (scratch assay)2.8 Evaluation of the in vitro inflammatory response in AGS cells3. Phytochemical Analysis3.1 Preliminary phytochemical assays3.2 Quantitative analysis of secondary metabolites3.3 Thin Layer Chromatography Analysis3.4 Analysis by High Performance Liquid Chromatography (HPLC)3.5 Mass spectrometry3.6 Electrospray Ionization Mass Spectrometry (ESI-MS) analysis3.7 Identification of phenolic compounds by HPLC-LC-ESI- MS/MS3.8 Quantitative determination of total alkaloids content by HPLC3.9 Quantitative determination of total alkaloids content by HPLC4. Preclinical Drug Dose Calculation4.1 Conversion based on normalization of dose-to body surface area4.2 Determination of animal equivalent dose5. Toxicology Studies (In Vitro and In Vivo)5.1 Cytotoxicity evaluation using Alamar blue5.2 Micronucleus test5.3 Comet test5.4 Effect on cell cycle5.5 In vivo acute toxicity study5.6 Subchronic toxicity (repeated doses)6. Standard Procedure for Anesthesia in Preclinical Experiments6.1 Planning and anesthetic care6.2 Pre-Procedure6.3 Experimental Procedure7. General Considerations and Collection of Animal Blood7.1 Vein blood collection7.2 Tail vein blood collection7.3 Cardiac puncture blood collection7.4 Posterior vena cava7.5 Orbital sinus8. Animal Experiments on the Cardiovascular System8.1 Obtaining electrocardiogram (ECG) and blood pressure (BP) signals8.2 Myocardial ischemia induction9. Animal Experiments on Ulcerative Colitis9.1 Ulcerative colitis induced by 2,4,6-Trinitrobenzenesulfonic acid (TNBS)9.2 Chronic ulcerative colitis induced by Dextran Sodium Sulfate (SSD) 2%9.3 Colitis with recurrence (Chronic)10. Experiments on Anti-Bacterial Activities10.1 Antibacterial activity by in vitro assays10.2 Animal Experiments on antibacterial activity11. Experiments on Anti-Fungal Activities11.1 Agar Disc Diffusion Method11.2 Minimum Inhibitory Concentration (MIC)11.3 Sorbital test11.4 Neurospora crassa assay11.5 Effect of Ergosterol on MIC11.6 Test of 1,3-D-Glucan Synthase (GS)11.7 Virulence Factors Inhibition of Candida albicans11.8 Germinative Tube Inhibition Test11.9 Biofilm assay11.10 Time-kill studies11.11 Determination of Antifungal Activity by in vivo12. Animal Experiment on Wound Healing Activity12.1 Excision wound healing model12.2 Incision wound healing model12.3 Evaluation of the effect of the extract on migration/proliferation in fibroblasts12.4 Surgical wound infected in rats (in vivo)13. Animal Experiments of Anti-inflammatory activities13.1 Inflammatory Paw edema13.2 Lipopolysaccharide (LPS)-induced peritonitis13.3 Murine Model of Immediate Hypersensitivity Induced By OVA13.4 Air pouch inflammation model13.5 Cotton pellet inflammation model14. Animal Experiment on Anti-Nociceptive Activities14.1 Abdominal constriction by acetic acid14.2 Formalin test 2.5%14.3 Hot plate Test15. Experiments on Anti-Oxidant Activities15.1 DPPH free radical scavenging activity15.2 Hydrogen peroxide radical scavenging activity15.3 Free radical nitric oxide scavenging activity15.4 Ferric Reducing Antioxidant Power assay15.5 Determination of superoxide dismutase (SOD)15.6 Glutathione peroxidase (GPx) activity15.7 Determination of catalase (CAT)15.8 Determination of reduced glutathione (GSH)15.9 Determination of malonaldehyde (MDA)15.10 Total protein dosage15.11 Test of oxidative degradation of deoxyribose16. Animal Experiments on Hepatoprotective Activities16.1 Acute liver damage by paracetamol16.2 Treatment of acute ethanol-induced injury16.3 The induction of liver damage by carbon tetra chloride (CCl4)17. Animal Experiments on Anti-Diarrheal Activities17.1 Gastric emptying17.2 Intestinal transit17.3 Enter pooling induced by castor oil17.4 Microvascular permeability17.5 Activity on the H+/K+ pump - ATPaseAnimal Experiments on Central Nervous System (CNS)18.1 Stress Induced experiment18.2 Elevated Cross Maze (ECM)18.3 Open Field Test (OFT)18.4 Object Recognition Test (ORT)18.5 Murinometric evaluationExperiments on Anti-Cancer Activities (In Vitro and In Vivo)19.1 Cytotoxicity test19.2 Cell cycle assay19.3 Assessment of cell viability and proliferation using the MTT method19.4 Evaluation of clonogenic survival of different cell lines19.5 In vivo anti-tumor activityAnimal Experiments on Anti-Diabetic Activities20.1 Inhibition of -amylase 20.2 Inhibition of -glucosidase 20.3 In vivo Antidiabetic activityAnimal Experiments on Gastric Ulcer 21.1 Gastric Injury Induced by Ethanol21.2 Gastric Injury Induced by Acidified ethanol (Ethanol/HCl)21.3 Indomethacin induced gastric injury21.4 Gastric Injury Induced by Acetic Acid (chronic ulcer) in mice21.5 NSAID ulcer experiment induced by Piroxicam21.6 Pylorus ligation experiment in mice21.7 Stress-induced gastric injury (water restraint)21.8 Role of NO in a gastric ulcer model pretreated with L-NAME21.9 Gastric Injury Induced by Ethanol/HCl for quantification of mucus21.10 Ethanol/HCl-Induced Gastric Lesion with Indomethacin, L-NAME, Glibenclamide, Yohimbine21.11 Ischemia-reperfusion-induced gastric ulcer21.12 Evaluation of anti-H. pylori activity in vivo21.13 Evaluation of anti-Helicobacter pylori activity in vitroProtein Extraction and Western Blot Analysis22.1 Gel preparation22.2 Samples application22.3 Running22.4 Preparing Membranes 22.5 RevelationGel Electrophoresis and PCR Amplification23.1 DNA Extraction23.2 Dosage of RNA and DNA23.3 Solutions for Electrophoresis and Western blottingMolecular Docking Methods for Drug Design24.1 Biological activity calculation24.2 Molecular docking between the inhibitors and target proteins