Bioassays in Experimental and Preclinical Pharmacology

By Karuppusamy Arunachalam and Sreeja Puthanpura Sasidharan

1. Overview

2. Cell Culture Assays

2.1 Cell Cycle Assay: Propidium Iodide Marking

2.2 Apoptosis Assay: Annexin V/Propidium Iodide Marking

2.3 Cytotoxicity in Caco-2 cells by the Alamar blue method

2.4 Viability in Caco-2 cells stimulated with lipopolysaccharide (LPS)

2.5 Determination of Nitric oxide (NO) by in vitro experiment

2.6 Analysis of specific cytotoxicity against RAW 264.7 for determination anti-inflammatory activity in vitro

2.7 Scratching of Fibroblast cells (scratch assay)

2.8 Evaluation of the in vitro inflammatory response in AGS cells

3. Phytochemical Analysis

3.1 Preliminary phytochemical assays

3.2 Quantitative analysis of secondary metabolites

3.3 Thin Layer Chromatography Analysis

3.4 Analysis by High Performance Liquid Chromatography (HPLC)

3.5 Mass spectrometry

3.6 Electrospray Ionization Mass Spectrometry (ESI-MS) analysis

3.7 Identification of phenolic compounds by HPLC-LC-ESI- MS/MS

3.8 Quantitative determination of total alkaloids content by HPLC

3.9 Quantitative determination of total alkaloids content by HPLC

4. Preclinical Drug Dose Calculation

4.1 Conversion based on normalization of dose-to body surface area

4.2 Determination of animal equivalent dose

5. Toxicology Studies (In Vitro and In Vivo)

5.1 Cytotoxicity evaluation using Alamar blue

5.2 Micronucleus test

5.3 Comet test

5.4 Effect on cell cycle

5.5 In vivo acute toxicity study

5.6 Subchronic toxicity (repeated doses)

6. Standard Procedure for Anesthesia in Preclinical Experiments

6.1 Planning and anesthetic care

6.2 Pre-Procedure

6.3 Experimental Procedure

7. General Considerations and Collection of Animal Blood

7.1 Vein blood collection

7.2 Tail vein blood collection

7.3 Cardiac puncture blood collection

7.4 Posterior vena cava

7.5 Orbital sinus

8. Animal Experiments on the Cardiovascular System

8.1 Obtaining electrocardiogram (ECG) and blood pressure (BP) signals

8.2 Myocardial ischemia induction

9. Animal Experiments on Ulcerative Colitis

9.1 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 Activities

10.1 Antibacterial activity by in vitro assays

10.2 Animal Experiments on antibacterial activity

11. Experiments on Anti-Fungal Activities

11.1 Agar Disc Diffusion Method

11.2 Minimum Inhibitory Concentration (MIC)

11.3 Sorbital test

11.4 Neurospora crassa assay

11.5 Effect of Ergosterol on MIC

11.6 Test of Β1,3-D-Glucan Synthase (GS)

11.7

• Format: Hardcover
• Language: English
• ISBN: 1071612328
• ISBN13: 9781071612323
• Release Date: February 2021
• Publisher: Humana
• Length: 271 Pages
• Weight: 1.50 lbs.
• Dimensions: 0.7" x 7.7" x 10.2"