Introduction to Basics of Pharmacology and Toxicology: Volume 3 : Experimental Pharmacology : Research Methodology and Biostatistics (Introduction to Basics of Pharmacology and Toxicology, 3)

Foreword
Preface to Volume 1: General and Molecular Pharmacology: Principles of Drug Action (https://link.springer.com/book/10.1007/978...
Preface to Volume 2
Preface
Acknowledgments
Contents
Editors and Contributors
Part I: Experimental Pharmacology
	1: Experimental Methodologies Involved in the Discovery of Drugs
		1.1 Basic Principles of Drug Discovery
		1.2 In-silico Techniques
			1.2.1 Quantitative Structure-Activity Relationship (QSAR)
			1.2.2 2D-QSAR and 3D-QSAR
			1.2.3 Molecular Docking
			1.2.4 Virtual High Throughput Screening
			1.2.5 Pharmacophore Mapping
		1.3 In-vitro Techniques
			1.3.1 Absorption Assays
			1.3.2 Metabolic Stability Assays
			1.3.3 CYP450 Pathway Elucidation
			1.3.4 Inhibition Potential
			1.3.5 Induction Potential
			1.3.6 Metabolic Profiling
			1.3.7 Antisense Technology
		1.4 In-vivo Techniques
			1.4.1 Gene Knockout Animals
			1.4.2 Transgenic Animals
			1.4.3 Monoclonal Antibodies Mediated Gene Manipulation
		1.5 Assays
		Bibliography
	2: Laboratory Animals
		2.1 Introduction
		2.2 Mouse
			2.2.1 Scientific Name
			2.2.2 Description
			2.2.3 Salient Points of the Mouse as Experimental Models
			2.2.4 Outbred Stocks of the Mouse
			2.2.5 Inbred Strains of the Mouse
			2.2.6 Genetically Modified Mouse
			2.2.7 Limitations of the Mouse as Experimental Models for Human Diseases
		2.3 Rat
			2.3.1 Scientific Name
			2.3.2 Physical Description
			2.3.3 Salient Points of the Rat as an Experimental Animal
			2.3.4 Outbred Stocks of Rat
			2.3.5 Inbred Strains of Rat
			2.3.6 Transgenic and Knock-out Rats
			2.3.7 Limitation of the Rat as an Experimental Animal
		2.4 Guinea Pig
			2.4.1 Scientific Name
			2.4.2 Description
			2.4.3 Salient Points of the Guinea Pig as an Experimental Animal
			2.4.4 Strains of Guinea Pig
		2.5 Hamster
			2.5.1 Scientific Name
			2.5.2 Description
			2.5.3 Salient Points of Hamster as an Experimental Animal
		2.6 Gerbil
		2.7 Rabbit
		2.8 Cats
		2.9 Pig
		2.10 Dog
		2.11 Monkeys and Other Non-human Primates
		2.12 Horse
		2.13 Sheep
		2.14 Chicken
		2.15 Pigeon
		2.16 Frog
		2.17 Zebra Fish
		Bibliography
	3: Care and Handling of Laboratory Animals
		3.1 Introduction
		3.2 Animal Ethics
		3.3 Housing and Environment
		3.4 Handling of Commonly Used Animals in Biomedical Research
			3.4.1 Handling of Mice
			3.4.2 Handling of Rat
			3.4.3 Handling of Guinea Pig
			3.4.4 Handling of Rabbit
			3.4.5 Handling of Hamster
			3.4.6 Handling of Gerbils
			3.4.7 Handling of Ferret
			3.4.8 Handling of Dog
			3.4.9 Handling of Cat
		3.5 Animal Welfare
		Bibliography
	4: Biological Sample Collection from Experimental Animals
		4.1 Introduction
		4.2 Collection of Blood
			4.2.1 General Principles
			4.2.2 Blood Sample Collection Through Saphenous Vein
				Caution:
			4.2.3 Blood Sample Collection Through Dorsal Pedal Vein
				Caution:
			4.2.4 Blood Sample Collection Through Tail Vein
				Caution:
			4.2.5 Blood Sample Collection Using a Tail Bleed (Tail Snip and Tail Nick) Method
				Caution:
			4.2.6 Blood Sample Collection Through Orbital Sinus
				Caution:
			4.2.7 Blood Sample Collection Through Jugular Vein
				Caution:
			4.2.8 Blood Sample Collection Using a Temporary Cannula
			4.2.9 Tarsal Vein Blood Sample Collection
				Caution:
			4.2.10 Marginal Ear Vein/Artery Blood Sample Collection
			4.2.11 Blood Sample Collection Through Cephalic Vein
			4.2.12 Blood Sample Collection Through Cranial Vena Cava
			4.2.13 Blood Sample Collection Through Posterior Vena Cava
			4.2.14 Blood Sample Collection Through Cardiac Puncture
			4.2.15 Adverse Effects of Blood Sample Collections
		4.3 Urine Sample Collecting Methods
		4.4 Saliva Sample Collection Method
		4.5 Conclusion
		Bibliography
	5: Anesthesia and Euthanasia of Experimental Animals
		5.1 Anesthesia of Experimental Animals
			5.1.1 Anesthesia
			5.1.2 Stages of Anesthesia
			5.1.3 Pre-Anesthetic Medication
			5.1.4 Types of Anesthesia
				5.1.4.1 Local Anesthesia
				5.1.4.2 General Anesthesia
			5.1.5 Monitoring of Depth of Anesthesia
			5.1.6 Postoperative Analgesia
		5.2 Euthanasia of Experimental Animals
		Bibliography
	6: Equipments in Experimental Pharmacology
		6.1 Organ Bath
			6.1.1 History
			6.1.2 Contents
		6.2 Kymograph
			6.2.1 History
			6.2.2 Procedure
		6.3 Tissue Levers
			6.3.1 Magnification
			6.3.2 Levers Used in Experimental Pharmacology
		6.4 Analgesiometer
			6.4.1 Basic Principle
			6.4.2 Hot Plate Analgesiometer
			6.4.3 Procedure
		6.5 Digital Analgesiometer:Tail-Flick Method
			6.5.1 Procedure
		6.6 Electroconvulsiometer
			6.6.1 Principle
			6.6.2 Procedure
			6.6.3 Threshold for Electroconvulsions
		6.7 Elevated Plus-Maze
			6.7.1 Equipment
			6.7.2 Principle
			6.7.3 Procedure
		6.8 Actophotometer
			6.8.1 Principle
			6.8.2 Procedure
		6.9 Rota-Rod Apparatus
			6.9.1 Equipment
			6.9.2 Principle
			6.9.3 Procedure
		6.10 Grip Strength Meter
			6.10.1 Principle
			6.10.2 Procedure
		6.11 Cook´s Pole-Climbing Apparatus
			6.11.1 Principle
			6.11.2 Equipment
			6.11.3 Procedure
			6.11.4 Experiment
		Bibliography
	7: Bioassay and Experiments on Isolated Muscle Preparations
		7.1 Definition
		7.2 Classification
		7.3 Indications
		7.4 Principles
		7.5 Purpose
		7.6 Methods of Bioassay for Agonist
			7.6.1 Direct-End-Point Method
			7.6.2 Matching Method
			7.6.3 Graphical Method
			7.6.4 Multiple-Point Assays
		7.7 Standard Isolated Muscle Preparations
			7.7.1 Terminologies
			7.7.2 Muscle Preparations Used in Bioassay
		Bibliography
	8: Experiments on Anaesthetized Intact Animals
		8.1 Introduction
		8.2 Intact Animal Models n Dog
			8.2.1 Blood Pressure Recording in the Dog
			8.2.2 Spleen Volume Measurement
			8.2.3 Diuretic Study in Dogs
		8.3 Intact Animal Models in Cat
			8.3.1 Nictitating Membrane
			8.3.2 Spinalisation of the Cat
			8.3.3 Open Chest Preparation
			8.3.4 Muscle Nerve Preparations
			8.3.5 Models to Study the Action of Centrally Acting Skeletal Muscle Relaxants
			8.3.6 Hindquarters Perfusion Model
			8.3.7 Domenjoz Method for Antitussive Action
		8.4 Intact Animal Models in Guinea Pigs
			8.4.1 Konzett-Rossler Method for Bronchodilator Activity
		8.5 Intact Animal Models in Rats
			8.5.1 Blood Pressure Recording in Rats
		8.6 Conclusion
		Bibliography
	9: Bioassay of Standard Compounds
		9.1 Bioassay on Acetylcholine
		9.2 Bioassay of Adrenaline
		9.3 Bioassay of Histamine
		9.4 Bioassay of 5HT (Serotonin)
		9.5 Angiotensin II Bioassay
		9.6 Bioassay of Vasopressin
		9.7 Bioassay of ACTH
		9.8 Bioassay of Estrogen
		9.9 Bioassay of Oxytocin
		Bibliography
	10: Principles of EC50, ED50, pD2 and pA2 Values of Drugs
		10.1 The Concept of ``Dose Descriptors´´
		10.2 Preclinical Toxicity Studies
		Bibliography
	11: Physiological Salt Solutions
		11.1 Introduction
		11.2 General Principles in Preparation of Physiological Salt Solution
		11.3 Commonly Used Physiological Salt Solution
		11.4 Choice of Physiological Salt Solution for Specific Tissues
		Bibliography
	12: Validation of Animal Models
		12.1 Introduction
		12.2 Animal Model
			12.2.1 History -Animals in Research
			12.2.2 Classification of Animal Model
			12.2.3 Utilization of Animal Models in Scientific Field
			12.2.4 Ideal Animal Model
		12.3 Validation of Animal Model
			12.3.1 Face Validity
			12.3.2 Construct Validity
			12.3.3 Predictive Validity
			12.3.4 Internal Validity
			12.3.5 Factors Affecting Internal Validity
		12.4 Process of Developing an Animal Model
		12.5 Validation Process for Predictive Validity
		12.6 Framework to Identify Models of Disease (FIMD)
		12.7 Evaluation of Valid Animal Model
		12.8 Limitations of Animal Model
		12.9 Conclusion
		Bibliography
	13: Screening Methods for the Evaluation of Antidepressant Drugs
		13.1 Introduction
		13.2 Animal Models of Depression
		13.3 Criteria for Valid Animal Models of Depression
		13.4 Genetic Models
			13.4.1 Traditional Genetic Mouse Models
			13.4.2 Models Using Optogenetic Tools
			13.4.3 Selective Breeding
		13.5 Stress Exposure
			13.5.1 Models of Acute Stress
				13.5.1.1 Water Wheel Model
				13.5.1.2 Forced Swim Test
				13.5.1.3 Tail Suspension Test
				13.5.1.4 Learned Helplessness Test
			13.5.2 Chronic Stress Models
				13.5.2.1 Chronic Mild Stress Model (CMS)
				13.5.2.2 Chronic Restraint Stress Model
				13.5.2.3 Chronic Social Defeat Stress
			13.5.3 Isolation Induced Hyperactivity
			13.5.4 Early Life Stress Model
		13.6 Secondary or Iatrogenic Depression Models
			13.6.1 Hormones of HPA Axis
			13.6.2 Retinoic Acid Derivatives
			13.6.3 Cytokines and Immune System Dysregulation
		13.7 Pharmacological Models
			13.7.1 Reserpine Induced Hypothermia
			13.7.2 Amphetamine Potentiation
			13.7.3 Apomorphine Antagonism
			13.7.4 Psychostimulant Withdrawal Paradigm
		13.8 Miscellaneous
			13.8.1 Resident Intruder Paradigm in Rats
			13.8.2 Muricidal Behavior in Rats
			13.8.3 Olfactory Bulbectomy Model
			13.8.4 Disruption of Circadian Rhythm
		13.9 Behavioral Endpoints
		13.10 Conclusion
		Bibliography
	14: Screening Methods for the Evaluation of Antiepileptic Drugs
		14.1 Introduction
		14.2 In-vitro Methods
			14.2.1 GABAA Receptor Binding Measurement in Rat Brain
			14.2.2 GABAB Receptor Binding Measurement in Rat Brain
			14.2.3 GABA Uptake in the Cerebral Cortex of the Rat
			14.2.4 GABA Release in Hippocampal Slices of Rats
			14.2.5 [3H]-CPP Binding Assay for Glutamate Receptors in Rat Cerebral ortex
			14.2.6 [3H]-TCP Binding Assay for NMDA Receptor Complex in Rat Cerebral Cortex
			14.2.7 [H3]-Glycine Binding Assay for NMDA Receptor Complex in Rat Cerebral Cortex
			14.2.8 [H3]-Strychnine Binding Assay for the Strychnine-Sensitive-Glycine Receptor in Rat Cerebral Cortex
			14.2.9 [35S]-TBPS Binding Assay for Picrotoxin Site in GABA Receptor Complex in Rat Cerebral Cortex
			14.2.10 Other In Vitro Techniques
		14.3 In Vivo Techniques
			14.3.1 Electrically Induced Seizure Models
			14.3.2 Chemically Induced Seizure Models
			14.3.3 Seizures Induced by Surgery
				14.3.3.1 Seizures Induced by Focal Lesions
				14.3.3.2 Seizures Induced by Kindling
				14.3.3.3 Post Hypoxic Myoclonus in Rats
		14.4 Classification of Animal Models Based on the Type of Seizure Disorder
			14.4.1 Models Used for Status Epilepticus
			14.4.2 Model for Infantile Spasm
				14.4.2.1 Genetic Models of Chronic Infantile Spasms:
			14.4.3 Models for GTCS
			14.4.4 Models for Complex Partial Seizures
			14.4.5 Models for Absence Seizures
			14.4.6 Models for Simple Partial Seizures
		14.5 Genetic Models of Epilepsy
			14.5.1 Genetic Animals Models with Spontaneous Seizures
			14.5.2 Genetic Animal Models with Absence Like Seizures and Tonic Convulsions
			14.5.3 Genetic Animal Models with Reflex Seizures
		14.6 Conclusion
		Bibliography
	15: Screening Methods for the Evaluation of General Anaesthetics
		15.1 Introduction
		15.2 Screening of Intravenous Anaesthetic Agents
			15.2.1 Evaluation of Intravenous Anaesthetic Agents in Mice
			15.2.2 Electroencephalographic (EEG) Threshold Test in Rats
			15.2.3 Evaluation of Safety of Intravenous Anaesthetics in Rabbits
		15.3 Screening of Inhalational Anaesthetic Agents
			15.3.1 Screening of Volatile Anaesthetics in Mice or Rats
			15.3.2 Determination of Minimal Alveolar Anaesthetic Concentration (MAC) in Rats
			15.3.3 Evaluation of Safety of Inhalational Anaesthetics in Rats
		Bibliography
	16: Screening Methods for the Evaluation of Antipsychotic Drugs
		16.1 Introduction
		16.2 In-Vitro Studies
			16.2.1 D1 Receptor Assay Using [H]-SCH-23390 in Rat Brain
			16.2.2 D2 Receptor Assay Using 3[H]-Spiroperidol in Rat Brain
			16.2.3 D3 Receptor Assay Using 3[H]-Nafadotride in Rat Brain
			16.2.4 D4 Receptor Assay Using 3[H]-L-745-870
			16.2.5 In-Vitro Assay for Other Receptors Involved in the Evaluation of Antipsychotic Action
		16.3 Invivo: Behavioral Tests
			16.3.1 Golden Hamster Test
			16.3.2 Cotton Rat Test
			16.3.3 Artificial Hibernation in Rats
			16.3.4 Catalepsy in Rodents
			16.3.5 Pole-Climb Avoidance in Rats
			16.3.6 Brain Self-Stimulation Test in Rats
			16.3.7 Pre-Pulse Inhibition of Startle Response in Rats
			16.3.8 Other Behavioral Tests Used
		16.4 In vivo Test Based on Mechanism of Action
		16.5 Genetic Models of Psychosis
			16.5.1 Dominant-Negative DISC-1 Transgenic Mice
			16.5.2 Gsα-Transgenic Mice
			16.5.3 Sandy Mice or Dysbindin-1 Mutant (DTNBP-1) Knockout Mice
			16.5.4 NR-1 Hypomorphic (NR-1H) Mice
			16.5.5 Heterozygous Reeler Mouse
			16.5.6 Other Genetic Animal Models of Psychosis
		Bibliography
	17: Screening Methods for the Evaluation of Sedative-Hypnotics
		17.1 Introduction
		17.2 In-vitro Studies
			17.2.1 GABAA Receptor Binding Measurement in Rat Brain
			17.2.2 GABAB Receptor Binding Measurement in Rat Brain
			17.2.3 [3H]-Mesulergine Binding Assay for 5HT2C Receptors
			17.2.4 [3H]-Mepyramine Binding Assay for H1 Receptors
		17.3 In-Vivo Tests for Sedative Activity
			17.3.1 Effects on Motility (Sedative or Stimulatory Activity)
				17.3.1.1 Method of Intermittent Observations
				17.3.1.2 Open Field Test
				17.3.1.3 Hole-Board Test
				17.3.1.4 Combined Open Field Test
				17.3.1.5 EEG Analysis from Rat Brain by Telemetry
			17.3.2 Tests for Muscle Coordination
				17.3.2.1 Inclined Plane
				17.3.2.2 Chimney Test
				17.3.2.3 Grip Strength Test
				17.3.2.4 Rotarod Method: (Refer to Chap. 42)
		17.4 In-vivo Tests for Hypnotic Activity
			17.4.1 General Considerations
			17.4.2 Potentiation of Hexobarbital Sleeping Time
			17.4.3 Experimental Insomnia in Rats
			17.4.4 EEG Registration in Conscious Cats
			17.4.5 Automated Rat Sleep Analysis System
		Bibliography
	18: Screening Methods for the Evaluation of Anxiolytic Drugs
		18.1 Introduction
		18.2 Light/Dark Exploration Test
		18.3 Elevated Plus Maze Test
		18.4 Four-Plate-Test
		18.5 Fear Potentiated Startle Test
		18.6 Defensive Burying Test
		18.7 Social Interaction
		18.8 Separation-induced Ultrasonic ``Distress´´ Vocalization
		18.9 Vogel Conflict Test
		18.10 Staircase Test
		18.11 Hole-Board Test
		18.12 Open Field Test
		18.13 Conclusion and Future Perspectives
		Bibliography
	19: Screening Methods forthe Evaluation of AntiparkinsonianDrugs
		19.1 Introduction
		19.2 In vitro Techniques
			19.2.1 Primary Microglial Cultures
			19.2.2 Animal Striatal Slices
			19.2.3 Dopamine Receptors Assessment by Radioligand Binding
			19.2.4 Dopamine Transporter Assessment by Radioligand Binding
			19.2.5 Dopamine Release from Synaptosomes
			19.2.6 Assessment of Neuroprotective Efficacy
		19.3 In vivo Models
		19.4 Drug-Induced Animal Models of PD
			19.4.1 Reserpine Model of PD
			19.4.2 Methamphetamine Model of PD
			19.4.3 Neuroleptics Induced PD Models
			19.4.4 Cholinomimetics Induced PD Models
		19.5 Toxin Induced PD Models
			19.5.1 MPTP Models in Monkeys
			19.5.2 Rotenone Induced PD Models
			19.5.3 Paraquat Model for PD
			19.5.4 Maneb-Paraquat Model
			19.5.5 6-OHDA (6-Hydroxy Dopamine or Oxdopamine) Model of PD
			19.5.6 Manganese
		19.6 Genetic Models of PD
			19.6.1 α-Synuclein Transgenic Mouse Models
			19.6.2 α-Synuclein Transgenic Drosophila Model
			19.6.3 Parkin Transgenic Mouse Models
			19.6.4 Pink-1 Deleted Mutant Drosophila and Mice Model
			19.6.5 DJ-1 Deficient Transgenic Mouse/Drosophila Models
			19.6.6 LRRK2 Gene Knock-in Transgenic Mouse
			19.6.7 Mitopark Mice
		Bibliography
	20: Screening Methods for theEvaluation of Drugs for Learning and Memory
		20.1 Introduction
		20.2 In vitro Methods
			20.2.1 In vitro Inhibition of Acetylcholine-Esterase (AChE) Activity in Rat Striatum
			20.2.2 In vitro Inhibition of Butyrylcholine-Esterase (BChE) Activity in Human Serum
			20.2.3 In vitro Estimation of Presynaptic Release of [3H]- ACh and Other Transmitters from Rat Brain
			20.2.4 In-vitro [3H]-Oxotremorine-M Binding to a Muscarinic Cholinergic Receptor in Rat Forebrain
			20.2.5 In-vitro [3H]-N-Methylscopolamine Binding in Presence/Absence of Guanylyimidophosphate
			20.2.6 Transfected Cell Lines
			20.2.7 Evaluation of beta and gamma Secretase Inhibition
			20.2.8 Cultures of Rat Hippocampal and Cerebral Cortex Neurons
			20.2.9 Study of Field Excitatory Postsynaptic Potential
			20.2.10 Other in-vitro Techniques Used for Evaluating Activity in AD
		20.3 In-vivo Behavioral Models:Passive (Inhibitory) Avoidance Tests
			20.3.1 Step Down Model
			20.3.2 Step Through Model
			20.3.3 Uphill Avoidance Model
			20.3.4 Two-Compartment Test
			20.3.5 Trial to Criteria Inhibitory Avoidance
		20.4 In-vivo Behavioral Models: Active Avoidance Tests
			20.4.1 Runaway Avoidance Test
			20.4.2 Shuttle Box (Two-way) Avoidance Test
			20.4.3 Jumping (One-way Shuttle) Avoidance Test
		20.5 In-vivo Discrimination Learning Tests
			20.5.1 Spatial Habituation Learning
			20.5.2 Spatial Habituation Learning
			20.5.3 Spatial Learning in a Radial Arm
			20.5.4 Spatial Learning in the Water Maze (Morris Test)
			20.5.5 Visual Discrimination Test
			20.5.6 Olfactory Discrimination Test
		20.6 In-vivo Experiments in Other Animals
			20.6.1 Aversive Discrimination in Chickens
			20.6.2 Discrimination Studies in Aged Monkeys
		20.7 Genetic Models for Testing Memory and Learning
		20.8 Invertebrate Models
		Bibliography
	21: Screening Methods for the Evaluation of Antianginal Agents
		21.1 Introduction
		21.2 In-vitro Models
			21.2.1 Isolated Heart (Langendorff) Preparation
			21.2.2 Gottlieb and Magnus Method (Balloon Method)
			21.2.3 Doring and Dehnert (1998) Method
			21.2.4 Isolated Rabbit Aorta Preparation
			21.2.5 Calcium Antagonism in the Pitched Rat
			21.2.6 Relaxation of Bovine Coronary Artery
			21.2.7 Coronary Artery Ligation in Isolated Rat Heart
			21.2.8 Isolated Heart-Lung Preparation
			21.2.9 Plastic Cast Technique in Dogs
		21.3 In-vivo Methods
			21.3.1 Occlusion of Coronary Artery in Anesthetized Dogs and Pigs
			21.3.2 Microspheres Induced Acute Ischemia
			21.3.3 Isoproterenol-Induced Myocardial Necrosis in Rats
			21.3.4 Stenosis-Induced Coronary Thrombosis Model
			21.3.5 Electrical Stimulation-Induced Coronary Thrombosis
			21.3.6 Myocardial Ischemic Preconditioning Model
			21.3.7 Models of Coronary Flow Measurement
		Bibliography
	22: Screening Methods for the Evaluation of Antihypertensive Drugs
		22.1 Introduction
		22.2 Measurement of Blood Pressure in Animals
		22.3 Animal Models of Hypertension
		22.4 In-Vitro Models of Hypertension
			22.4.1 Endothelin Receptor Antagonism in Porcine Isolated Hearts
			22.4.2 Monocrotaline Induced Pulmonary Hypertension
			22.4.3 ACE inhibition in Isolated Guinea Pig Ileum
			22.4.4 Isolated Guinea Pig Atria
		22.5 Rat models of Hypertension
			22.5.1 Reno-Vascular Hypertension Models in Rats
				22.5.1.1 Two Kidney One Clip (2K1C) Hypertension [Goldblatt Hypertension Model]
				22.5.1.2 One Kidney One Clip Model (Chronic Renal Hypertension in Rats)
				22.5.1.3 Two-Kidney Two Clip Method [Chronic Renal Hypertension in Rats]
			22.5.2 Dietary Hypertension Models in Rats
				22.5.2.1 Fructose-Induced Hypertension in Rats
				22.5.2.2 High Salt-Induced Hypertension in Rats
			22.5.3 Neurogenic Hypertension Model in Rats
			22.5.4 Endocrine Hypertension
				22.5.4.1 DOCA Salt-Induced Hypertension in Rats
			22.5.5 Psychogenic Hypertension Models in Rats
			22.5.6 Genetic Models of Hypertension in Rats
		22.6 Dog Models of Hypertension
			22.6.1 Chronic Renal Hypertension
			22.6.2 Neurogenic Hypertension
		22.7 Monkey Models of Hypertension
			22.7.1 Renin Inhibition in Monkeys
		22.8 Transgenic Models
			22.8.1 Transgenic Rats Overexpressing Mouse Ren-2 Gene [TGR (mRen 2) 27]
			22.8.2 Knockout Models
		Bibliography
	23: Screening Methods for the Evaluation of Antiarrhythmic Drugs
		23.1 Introduction
		23.2 In Vitro Models
			23.2.1 Isolated Guinea Pig Papillary Muscle
			23.2.2 Langendorff Technique
			23.2.3 Acetylcholine or Potassium Induced Arrhythmias
		23.3 In-Vivo Models
			23.3.1 Chemically Induced Arrhythmias
			23.3.2 Electrically Induced Arrhythmias
			23.3.3 Exercise-Induced Ventricular Fibrillation
			23.3.4 Mechanically Induced Arrhythmias
			23.3.5 Genetically Induced Arrhythmia
		Bibliography
	24: Screening Methods for the Evaluation of Cardiotonic Drugs
		24.1 In vitro and Ex vivoMethods
			24.1.1 Ouabain Binding Assays
			24.1.2 Isolated Hamster Hearts
			24.1.3 Isolated Cat Papillary Muscle
		24.2 In Vivo Methods
			24.2.1 Mouse Models
			24.2.2 Rat models
				24.2.2.1 Models of Myocardial Injury
				24.2.2.2 Rat Left Coronary ArteryLigation Model
				24.2.2.3 Rat Aortic Banding Model (Pressure Overload Model)
				24.2.2.4 Dahl Salt-Sensitive Rats
				24.2.2.5 Spontaneous Hypertensive Rats and Spontaneous Hypertensive-Heart Failure Rats (SH-HF Rats)
		24.3 Large Animal Models
			24.3.1 Dog Models
				24.3.1.1 Chronic Rapid Pacing Model
				24.3.1.2 Volume Overload Model
				24.3.1.3 Pressure Overload Model
				24.3.1.4 Coronary Artery Ligation and Microembolization Model
			24.3.2 Rabbit Models
				24.3.2.1 Volume and Pressure Overload Model
				24.3.2.2 Tachycardia Pacing Model
				24.3.2.3 Doxorubicin Cardiomyopathy Model
			24.3.3 Guinea Pig Models
				24.3.3.1 Cardiac Insufficiency Model
				24.3.3.2 Heart Failure Associated with Sudden Cardiac Death
				24.3.3.3 DOCA-Salt Associated Heart Failure
			24.3.4 Syrian Hamster Models
				24.3.4.1 Cardiomyopathic Syrian Hamsters
		24.4 Genetic Mice Models
			24.4.1 Myosin Lim Protein Knockout Mice
			24.4.2 TNF-Alpha Overexpressing Mice
		24.5 Newer Target Identification in CHF and Other Modalities of Treatment
		Bibliography
	25: Screening Methods for the Evaluation of Antiplatelet Drugs
		25.1 Introduction
		25.2 Targets for the Antiplatelet Drug
		25.3 Antiplatelet Drugs
		25.4 Screening for Antiplatelet Drugs
			25.4.1 Invitro Screening Method
				25.4.1.1 Plate Aggregation in Platelet-Rich Plasma (PRP) or Washed Platelets (WP)
				25.4.1.2 Platelet Aggregation by Laser Scattering Mmethod
				25.4.1.3 Platelet Aggregation After Gel Filtration
				25.4.1.4 Microchannel Array Flow Analyzer (MC-FAN)
				25.4.1.5 Platelet Function Aanalyzer-100 (PFA-100)
			25.4.2 In-Vivo Models
				25.4.2.1 Chemically Induced Thrombosis Model
					FeCl3 Induced Thrombosis
				25.4.2.2 Photochemical Model for Thrombosis
				25.4.2.3 Rose Bengal Induced Thrombosis
				25.4.2.4 Laser-Induced Thrombosis Model
				25.4.2.5 Arteriovenous Shunt Model for Thrombosis
				25.4.2.6 Electrical Induced Thrombosis Model
				25.4.2.7 Mechanically Induced Thrombosis Model/Stenosis Induced Thrombosis Model
			25.4.3 Zebrafish as a Thrombosis Model
			25.4.4 Genetic Models
		Bibliography
	26: Screening Methods for the Evaluation of Diuretics
		26.1 Introduction
		26.2 In Vitro Methods
			26.2.1 Carbonic Anhydrase Inhibition In Vitro
				26.2.1.1 Principle
				26.2.1.2 Procedure
				26.2.1.3 Evaluation
			26.2.2 Patch-Clamp Technique in Kidney Cells
				26.2.2.1 Principle
				26.2.2.2 Procedure
				26.2.2.3 Evaluation
			26.2.3 Perfusion of Isolated Kidney Tubules
				26.2.3.1 Principle:
				26.2.3.2 Procedure
				26.2.3.3 Evaluation
			26.2.4 Isolated Perfused Kidney
				26.2.4.1 Principle
				26.2.4.2 Procedure
				26.2.4.3 Evaluation
		26.3 In Vivo Methods
			26.3.1 Diuretic Activity in Rats (LIPSCHITZ Test) (Refer Fig. 26.2)
				26.3.1.1 Principle
				26.3.1.2 Procedure
				26.3.1.3 Evaluation
			26.3.2 Saluretic Activity in Rats
				26.3.2.1 Principle
				26.3.2.2 Procedure
				26.3.2.3 Evaluation
			26.3.3 Micropuncture Technique in the Rat (Refer Fig. 26.3)
				26.3.3.1 Principle:
				26.3.3.2 Procedure:
				26.3.3.3 Evaluation
			26.3.4 Diuretic and Saluretic Activity in Dogs (Refer Fig. 26.4)
				26.3.4.1 Principle
				26.3.4.2 Procedure:
				26.3.4.3 Evaluation:
			26.3.5 Clearance Methods (Refer Fig. 26.5)
				26.3.5.1 Principle (Refer Table 26.5):
				26.3.5.2 Procedure
				26.3.5.3 Evaluation:
			26.3.6 Stop-Flow Technique (Refer Fig. 26.6)
				26.3.6.1 Principle
				26.3.6.2 Procedure
				26.3.6.3 Evaluation
		Bibliography
	27: Screening Methods for the Evaluation of Antihyperlipidemic Drugs
		27.1 Lacunae with Current Anti-Hyperlipidemics
		27.2 How to Choose a Relevant Model for Screening?
		27.3 Classification of Screening Methods for Anti-Hyperlipidemias
			27.3.1 In-Vitro Screening Methods
				27.3.1.1 Inhibition of Isolated HMG-CoA Reductase
				27.3.1.2 In Vitro Assay Using Caco-2 Cell Lines
				27.3.1.3 ACAT Inhibitory Model
			27.3.2 In-Vivo Methods
				27.3.2.1 Triton-Induced Hyperlipidemic Rat Model
				27.3.2.2 Cholesterol Diet-Induced Hyperlipidemia
				27.3.2.3 Chronic Model or High-Fat Diet (HFD) Induced Hyperlipidemia
				27.3.2.4 Propylthiouracil (PTU) Induced Hyperlipidemia
				27.3.2.5 Fructose Induced Hyperlipidemia in Rats
				27.3.2.6 Hypolipidemic Activity in Syrian Hamsters
				27.3.2.7 Hereditary Hyperlipidemia in Rats and Rabbits
				27.3.2.8 Transgenic Animal Models
		Bibliography
	28: Screening Methods for the Evaluation of Antiulcer Drugs
		28.1 Introduction
		28.2 In Vitro Methods
			28.2.1 Neutralization Effect on Artificial Gastric Acid
			28.2.2 Vatier´s Artificial Stomach Apparatus Model
			28.2.3 Fordtran´s Model
		28.3 In Vivo Method: Gastric Ulcer Evaluation
			28.3.1 Acute Gastric Ulcer Evaluation
				28.3.1.1 Rats: Pylorus Ligation Method (Shay Rat Model)
				28.3.1.2 Stress Ulcer Model in Rats: Restrain-Induced Ulcers (Immobilization Stress), Hanson and Brodie
				28.3.1.3 Stress Ulcer Model in Rats: Cold Water Immersion Restraint (WRS) Stress, Takagi et al
				28.3.1.4 Stress Ulcer Model in Rats: Cold Restraint stress, Senay and Levine (1967); Das et al (1993)
				28.3.1.5 Stress Ulcer Model in Rats: Stress and NSAID-Induced Ulcers
				28.3.1.6 Stress Ulcer Model in Rats: Haemorrhagic Shock-Induced Gastric Ulcers
				28.3.1.7 Histamine-Induced Ulcer in Guinea Pigs
				28.3.1.8 NSAIDs-Induced Gastric Ulcer
				28.3.1.9 Ethanol-Induced Mucosal Damage in Rats
			28.3.2 Sub-Acute Gastric Ulcer Evaluation
			28.3.3 Chronic Gastric Ulcer: Acetic Acid-Induced Gastric Ulcer
		28.4 In Vivo Method: Duodenal Ulcer Evaluation
			28.4.1 Cysteamine-Induced Duodenal Ulcers
			28.4.2 Chemical-Induced Duodenal Ulcers
		Bibliography
	29: Screening Methods for the Evaluation of Antidiarrheal Drugs
		29.1 Introduction
		29.2 In Vitro Methods
		29.3 In-Vivo Methods
			29.3.1 Chemical-Induced Diarrhoea
				29.3.1.1 Castor Oil-Induced Diarrhoea
				29.3.1.2 Magnesium Sulphate Induced Diarrhoea
				29.3.1.3 Serotonin Induced Diarrhoea
			29.3.2 Chemical-Induced Enteropooling
				29.3.2.1 Castor Oil-Induced Enteropooling
				29.3.2.2 PGE2 Induced Enteropooling
			29.3.3 Gastrointestinal Transit Time Using Charcoal Meal
			29.3.4 Antidiarrheal Effect in Cecectomized Rats
			29.3.5 Antidiarrheal Effect in Cold-Restrained Rats
		Bibliography
	Untitled
	30: Screening Methods for the Evaluation of Antiemetics
		30.1 Introduction
		30.2 In-Vitro Methods
		30.3 In-Vivo Methods
			30.3.1 Chemicals/Drugs Induced Emesis Model
				30.3.1.1 Apomorphine Induced Emesis
				30.3.1.2 Copper Sulphate Induced Emesis
				30.3.1.3 Cisplatin-Induced Emesis
				30.3.1.4 Cisplatin-Induced Delayed Emesis
				30.3.1.5 Methotrexate Induced Delayed Emesis
				30.3.1.6 Other models of drug-induced emesis
			30.3.2 Emesis Induced by Motion
			30.3.3 Emesis Induced by Radiation
		Bibliography
	31: Screening Methods for the Evaluation of Hepatoprotective Agents
		31.1 Introduction
		31.2 Classification of Liver Diseases
		31.3 Causes of Liver Disease
			31.3.1 Hepatotoxins
			31.3.2 General Mechanism of Action of Hepatotoxins
		31.4 Liver Function Tests
		31.5 Hepatoprotective Agents
		31.6 Screening Models
			31.6.1 In Vitro Models
			31.6.2 Ex Vivo Models
			31.6.3 In Vivo Models
		31.7 Evaluation of Hepatoprotective Effect
		31.8 Conclusion
		Bibliography
	32: Screening Methods for the Evaluation of Antiobesity Drugs
		32.1 Introduction
		32.2 Animal Models for Inducing Obesity
			32.2.1 High Fat Diet-Induced Obesity
			32.2.2 Cafeteria Diet-Induced Obesity
			32.2.3 Fat or Sugar Choice Diet-Induced Obesity
			32.2.4 Meal Feeding-Induced Obesity
			32.2.5 Surgically-Induced Hypothalamic Obesity
			32.2.6 Ovariectomy-Induced Obesity in Rat
			32.2.7 Castration-Induced Obesity in Male Rat
			32.2.8 Gold-Thioglucose-Induced Hypothalamic Obesity
			32.2.9 Monosodium Glutamate-Induced Obesity
			32.2.10 Drug-Induced Obesity
		32.3 Genetically Obese Animals
		32.4 Uncommon Models Used in Obesity
			32.4.1 Obesity Induction in Macaques
			32.4.2 Drosophila Model for Obesity
		32.5 In-vitro Assay Methods for Evaluation of Anti-Obesity Activity
			32.5.1 Measurement of GDP Binding in Rat´s Brown Adipose Tissue
			32.5.2 Measurement of Uncoupling Protein-1 in Rat´s Brown Adipose Tissue
			32.5.3 β3-Adrenoceptor Binding in the Chinese Hamster Ovary Cells
			32.5.4 Determination of mRNA Level Of Leptin in Rat´s Adipose Tissue
			32.5.5 Plasma Leptin Level Determination in Rats
			32.5.6 Binding Assay of Neuropeptide Y (NPY) Receptor in Pigs
			32.5.7 Binding Assay of Orexin-A and Orexin B Receptors In Hamster Ovary Cells
			32.5.8 Binding Assay for Galanin Receptors in Rats
			32.5.9 Other In-vitro Assays
		32.6 In-Vivo Methods for Evaluation of Anti-obesity Activity
			32.6.1 Measuring Resting Metabolic Rate in the Mouse
			32.6.2 Measurement of Food Consumption in Rats
		Bibliography
	33: Absorption Studies
		33.1 Introduction
		33.2 Importance of Drug Absorption
		33.3 Classification of Drug Absorption Studies
		33.4 In vitro Methods to Evaluate Drug Absorption
			33.4.1 Partition Coefficient
				33.4.1.1 Log-P
				33.4.1.2 Log-D
			33.4.2 Parallel Artificial Membrane Permeability Assay (PAMPA)
			33.4.3 Brush Border Membrane Vesicles (BBMV)
			33.4.4 Isolated Intestinal Cells
			33.4.5 Everted Small Intestinal Sac Technique
			33.4.6 Everted Sac Modification Technique
			33.4.7 Cell Culture Techniques
		33.5 In-Vivo Models to Evaluate Drug Absorption
			33.5.1 Doluisio Method
			33.5.2 Intestinal Loop Technique
		33.6 In-Situ Method to Evaluate Drug Absorption
		33.7 Conclusion
		Bibliography
	34: Screening Methods for the Evaluation of Antiasthmatic Agents
		34.1 Introduction
		34.2 In-Vitro Models
			34.2.1 Checking for Smooth Muscle Relaxant Action
			34.2.2 Testing Anti-inflammatory Action
		34.3 Animal Models
			34.3.1 The Murine Allergic Airway Inflammation Model
			34.3.2 Other Animal Models
		34.4 Measuring Outcomes
		34.5 Example of Published Trial Testing Anti-muscarinic in an Acute Model of Asthma
		34.6 Example of Published Trial Testing a Chronic Model for Late-Phase Response
		Bibliography
	35: Screening Methods for the Evaluation of Antitussives and Expectorants
		35.1 Introduction
		35.2 Cough Induced by Chemical Stimuli
			35.2.1 Citric Acid-Induced Coughing in Guinea Pigs
		35.3 Cough Induced by Mechanical Stimuli
		35.4 Cough Induced by Stimulation of Superior Laryngeal Nerve
		35.5 Screening of Expectorants
			35.5.1 In Vitro Studies of Mucus Secretion
			35.5.2 In Vivo Studies of Mucus Secretion
				35.5.2.1 Acute Studies
				35.5.2.2 Chronic Cannulation Studies
		Bibliography
	36: Screening Methods for the Evaluation of Antidiabetic Drugs
		36.1 Introduction
		36.2 In-Vivo Models for T1DM
			36.2.1 Chemical Models for T1DM
				36.2.1.1 Streptozotocin-Based model
				36.2.1.2 Alloxan-Based Model
				36.2.1.3 Other Chemical Models
			36.2.2 Hormonal Models for T1DM
			36.2.3 Antibody-Mediated Models for T1DM
			36.2.4 Microbiological Models for T1DM
			36.2.5 Surgical Models for T1DM
			36.2.6 Genetic Models for T1DM
		36.3 In-Vivo Models for T2DM
			36.3.1 Chemical Models for T2DM
			36.3.2 Genetic Models for T2DM
				36.3.2.1 Monogenic Models
				36.3.2.2 Polygenic Models
				36.3.2.3 Transgenic and Knockout Models
				36.3.2.4 Miscellaneous Models
		36.4 Models for Diabetic Complications
			36.4.1 Models for Diabetic Nephropathy
			36.4.2 Models for Diabetic Retinopathy
			36.4.3 Models for Diabetic Neuropathy
			36.4.4 Models for Diabetic Cardiac Disease
		36.5 In-Vitro Models
		Bibliography
	37: Screening Methods for the Evaluation of Antithyroid Drugs
		37.1 Introduction
		37.2 Thyroid Hormones
			37.2.1 Historical Assays
			37.2.2 In Vitro Tests for Thyroid Hormones
			37.2.3 In Vivo Tests for Thyroid Hormones
		37.3 Antithyroid Drugs
			37.3.1 In Vivo Model for Antithyroid Activity
			37.3.2 Antithyroidal Effects in Animal Assays
		37.4 Conclusion
		Bibliography
	38: Screening Methods for theEvaluation of Antifertility Drugs
		38.1 Introduction
		38.2 Physiology of Female Sexual Organs
			38.2.1 Follicular Phase or Proliferative phase
			38.2.2 Luteal or Secretory Phase
		38.3 Estrous Cycle in Rodents
		38.4 Experimental Animal Models
			38.4.1 Invivo Methods
				38.4.1.1 Methods for Females
					Test for Anti-Ovulatory Activity
			38.4.2 Test for Estrogenic Activity
			38.4.3 Test for Anti-Estrogenic Activity
			38.4.4 Test for Progestational Activity
				38.4.4.1 Clauberg-McPhail Method
				38.4.4.2 Endometrial Carbonic Anhydrase Activity
				38.4.4.3 Deciduoma Reaction in Rats
				38.4.4.4 Assays causing changes in ovulation
				38.4.4.5 Pregnancy Maintenance Test
			38.4.5 Test for Anti-Progestational Activity
			38.4.6 Test for Anti-Implantation Activity /Abortifacient Activity
			38.4.7 Methods for Males.
			38.4.8 Test for Androgenic and Anabolic Activity:
		38.5 Test for Anti-Androgenic Activity
		38.6 Invitro Methods
			38.6.1 Methods for Females
				38.6.1.1 Test for Estrogenic Activity
				38.6.1.2 Techniques used to determine various parameters
			38.6.2 Test for Anti-Estrogenic Activity
			38.6.3 Test for Progestational Activity
			38.6.4 Methods for Males
				38.6.4.1 Emergent Spermatozoa Made Non-Functional
			38.6.5 Test for Anti-Androgenic Activity
		Appendix
			Phases of Menstrual Cycle
			Vaginal Opening Assay.
			Appearance of the vagina in different phases of estrous cycle of a Swiss albino strain mouse. a-Proestrus, b-Estrus, c- Metest...
		Bibliography
	39: Screening Methods for the Evaluation of the Drugs Acting on Posterior Pituitary, Adrenal Steroid, Testicular, Parathyroid,...
		39.1 Screening of Drugs Acting on the Thyroid System
			39.1.1 Tadpole Tail Culture Method
			39.1.2 Thyroidectomy Model
			39.1.3 Iodine Release Inhibition
			39.1.4 Anti-Goitrogenic Activity
			39.1.5 Reduction in Tensile Strength
		39.2 Screening of Drugs Acting on the Androgenic System (Table 39.2)
			39.2.1 Sebum Secretion in Rats
			39.2.2 Castration Technique
			39.2.3 Chicken Comb Model
			39.2.4 Nitrogen Retention Assay
			39.2.5 Growth of Secondary Sex Organs
			39.2.6 Change in Beta-glucuronidase Activity
		39.3 Screening of Drugs Acting on the Estrogenic System (Table 39.3)
			39.3.1 Castration of Female Rats
			39.3.2 Vaginal Cornification
			39.3.3 Increased Uterine Weight
			39.3.4 Chick Oviduct Method
		39.4 Screening of Drugs Acting on Progesterone System (Table 39.4)
			39.4.1 McPhail/Clauberg Test
			39.4.2 Rat Uterine C3 Model
			39.4.3 Rat Decidualization Test
			39.4.4 Rat Ovulation Inhibitionspiepr146 model
		39.5 Screening of Drugs Acting on Posterior Pituitary/Oxytocin System
			39.5.1 Milk Ejection Method
			39.5.2 Chicken Blood Pressure Method
			39.5.3 Isolated Uterus Method
		39.6 Screening of Drugs Acting on the Adrenal System
			39.6.1 Screening of Glucocorticoids (Table 39.5)
				39.6.1.1 Adrenalectomy in Rats
				39.6.1.2 Atrophy of Adrenal, Thymus Gland, and Spleen and Reduced Lymphocyte Count in Rats
			39.6.2 Screening of Mineralocorticoids (Table 39.6)
				39.6.2.1 Electrolyte Excretion
		39.7 Screening of Drugs Acting on the Parathyroid System (Table 39.7)
			39.7.1 Serum Calcium Increase Model
			39.7.2 Serum Phosphate Decrease Model
			39.7.3 cAMP Release Model
		Bibliography
	40: Screening Methods for the Evaluation of Analgesics, Anti-Inflammatory Drugs, and Antipyretics
		40.1 Screening for Analgesic Activity
			40.1.1 In Vitro Models for Analgesic Screening
				40.1.1.1 3H-Naloxone Binding Assay
				40.1.1.2 3H-Dihydromorphine Binding Assay
				40.1.1.3 3H-Bremazocine Binding Assay
				40.1.1.4 Enkephalinase Inhibition Assay
				40.1.1.5 Nociceptin Receptor Binding Assay
				40.1.1.6 Cannabinoid Receptor Binding Assay
				40.1.1.7 Vanilloid Receptor Binding Assay
				40.1.1.8 VIP Receptor Binding Assay
				40.1.1.9 Bioassay Techniques for Peripheral Mediators of Central Analgesia
			40.1.2 In Vivo Models for Analgesic Screening
				40.1.2.1 In Vivo Models for Central Analgesic Activity
				40.1.2.2 In Vivo Models for Peripheral Analgesic Activity
		40.2 Screening for Anti-Inflammatory Activity
			40.2.1 In Vitro Methods for Anti-Inflammatory Screening
			40.2.2 In Vivo Methods for Anti-Inflammatory Screening
				40.2.2.1 In Vivo Methods for Acute and Subacute Inflammation
			40.2.3 In Vivo Methods for Chronic Inflammation
			40.2.4 Miscellaneous Models of Anti-Inflammatory Activity
		40.3 Screening for Antipyretic Activity
			40.3.1 Brewer´s Yeast Suspension Method
			40.3.2 Bacterial Lipopolysaccharide Method
			40.3.3 Other Less Commonly Used Methods
		Bibliography
	41: Screening Methods for the Evaluation of Antiglaucoma and Anticataract Drugs
		41.1 Introduction
		41.2 Glaucoma
			41.2.1 Types of Glaucoma
			41.2.2 Aqueous Humor Outflow
			41.2.3 Intraocular Pressure Measurement
			41.2.4 Designing the Study to Test the Drug Efficacy
			41.2.5 Animal Models
		41.3 Cataract
			41.3.1 Animal Models of Congenital Cataracts
			41.3.2 Animal Models: Induced Cataract
			41.3.3 Hereditary Animal Models
			41.3.4 Diabetic-Induced cataracts
		41.4 Conclusion
		Bibliography
	42: Screening Methods for the Evaluation of Drugs Affecting Peripheral Nerve Functions
		42.1 Screening of Skeletal Muscle Relaxants (Neuromuscular Blocking Agents)
			42.1.1 Scope and Limitations of Neuromuscular Screening Tests
			42.1.2 In-Vitro Screening Tests
				42.1.2.1 Isolated Nerve-Muscle Preparations
				42.1.2.2 Single Muscle Fiber Study
				42.1.2.3 Ionophoretic Testing Method
				42.1.2.4 Human In Vitro Preparations
			42.1.3 Screening Tests in Anesthetized Animals
			42.1.4 Screening Tests in Intact Non-Anesthetized Animals
				42.1.4.1 Inclined Plane Method
				42.1.4.2 Rotarod Method
				42.1.4.3 Testing of Righting Reflexes
				42.1.4.4 Head Drop Technique
				42.1.4.5 Others
			42.1.5 Screening Tests in Anesthetized Human Beings
			42.1.6 Screening Tests in Intact Non-Anesthetized Human Beings
		42.2 Screening of Local Anesthetics
			42.2.1 Screening for Surface Anesthesia
			42.2.2 Screening for Infiltration Anesthesia
			42.2.3 Screening for Conduction Anesthesia
			42.2.4 Screening for Spinal and Epidural Anesthesia
		Bibliography
	43: Mutagenic Toxicity Testing
		43.1 Introduction
		43.2 History of Genotoxicity Testing
		43.3 Introduction to Genetic Toxicology Assays
			43.3.1 Metabolic Activation System
			43.3.2 Genotoxicity Testing Endpoints
		43.4 Predictors of Genotoxicity: Structural Alerts and In Silico Assays
		43.5 Assays for DNA Damage and Repair
			43.5.1 Comet Assay for DNA Strand Breakage
			43.5.2 DNA Repair Assays
				43.5.2.1 In Bacteria
				43.5.2.2 Unscheduled DNA Synthesis in Cultured Mammalian Cells and Intact Animals
			43.5.3 Assays Based on DNA Stress Response Mechanisms
				43.5.3.1 GreenScreen Assay or GADD45a-GFP Assay
		43.6 Gene Mutation Assays in Prokaryotes
		43.7 Assays in Nonmammalian Eukaryotes
			43.7.1 Sex-Linked Recessive Lethal Test
			43.7.2 Plant Assays
			43.7.3 Mitotic Recombination in Yeast
		43.8 Mammaliangene Mutation Assays
			43.8.1 In Vitro Assays for Gene Mutation in Mammalian Cells
			43.8.2 In Vivo Assays for Gene Mutation in Mammalian Cells
				43.8.2.1 Mouse Spot Test
				43.8.2.2 In Vivo HPRT Assay
				43.8.2.3 Pig-a Mutation Assay
			43.8.3 Transgenic Assays
		43.9 Mammalian Cytogenetic Assays
			43.9.1 Chromosome Aberrations
				43.9.1.1 In Vitro Mammalian Chromosome Aberration Test
			43.9.2 In Vivo Mammalian Chromosome Aberration Test
			43.9.3 Micronucleus Assay
			43.9.4 Sister Chromatid Exchange
			43.9.5 Aneuploidy
		43.10 Germ Cell Mutagenesis Assays
			43.10.1 Mouse Specific-Locus Test
			43.10.2 Cytogenetic Assays
			43.10.3 Mouse Heritable Translocation Assay
			43.10.4 Dominant Lethal Assay
			43.10.5 Assays for DNA Damage
		43.11 Regulatory Guidelines for Genotoxicity Testing
		43.12 Conclusion
		Bibliography
	44: Screening Methods for the Evaluation of Anticancer Drugs
		44.1 Introduction
		44.2 In-Vitro Methods
			44.2.1 Functional Assays (Cell Viability Assays)
			44.2.2 Non-Functional Assays (Morphological Assays)
				44.2.2.1 Assessment of Mode of Cell Death
				44.2.2.2 Assessment of Cell Death Markers
				44.2.2.3 Assessment of Genotoxicity of Compounds
			44.2.3 Molecular Techniques in In-Vitro Screening
		44.3 In-Vivo Screening Methods for Anticancer Drugs
			44.3.1 Chemical Carcinogen Models
				Box 44.1. Procarcinogen DMH Is Most Commonly Used for Inducing Colorectal Tumors in Rats and Mice
			44.3.2 Viral Infection Models
			44.3.3 Methods Involving Cell Line/Tumor Pieces Implantation
			44.3.4 Genetically Engineered Mouse Models
		Bibliography
	45: Screening Methods for the Evaluation of Drugs for Benign Prostatic Hyperplasia
		45.1 Introduction
		45.2 Pathophysiology and Pharmacology of BPH in Brief
		45.3 In Vitro Models
		45.4 In Vivo Models
			45.4.1 Testosterone Induced Benign Prostatic Hyperplasia in Rats
			45.4.2 Testosterone + Estrogen Induced Benign Prostatic Hyperplasia in Rats
			45.4.3 Sulpiride-Induced BPH in Rats
			45.4.4 Bladder Outlet Obstruction of Hemorrhagic Cystitis by Cyclophosphamide in Rats
			45.4.5 Single Dose of Cyclophosphamide Induced Hemorrhagic Cystitis in Rats of Bladder Outlet Obstruction Model
			45.4.6 Partially Ligated Urethra of Bladder Outlet Model
			45.4.7 Intraprostatic Injection Model
		45.5 Advantages of In Vivo Models
		45.6 Limitations of In Vivo Models
		45.7 Ex Vivo Models
			45.7.1 Isolated Prostate Gland Contractility Model (Flow Chart Given Below)
			45.7.2 Isolated Detrusor Muscle and Urethra Contractility Model
		45.8 Advantages of Ex Vivo Models
		45.9 Limitations With the Use of Ex Vivo Studies
		45.10 Transgenic Models
			45.10.1 Probasin Prolactin Transgenic Mouse Model of Benign Prostate Hyperplasia
			45.10.2 MMTV (Murine Mammary Tumor Virus) -Int2 (Proto-Oncogene) Transgenic Mice Model
			45.10.3 MMTV-Keratinocyte Growth Factor (fgf7)-Induced Hyperplasia of the Transgenic Model
		45.11 Advantages of Transgenic Models
		45.12 Limitations of Transgenic Models
		45.13 Xenograft Models
		45.14 Advantages With the Use of Xenograft Models
		45.15 Limitations with the Use of Xenograft Models
		45.16 Spontaneous BPH Model
		45.17 Conclusion
		Bibliography
	46: Screening Methods for the Evaluation of Drugs Acting on Autonomic Nervous System
		46.1 Introduction
		46.2 Screening of Unknown Substance for ANS Activity (Irwin Method)
		46.3 Sympathetic System
			46.3.1 In-Vitro Assays
			46.3.2 Isolated Organ Models
			46.3.3 In Vivo Models
		46.4 Parasympathetic System
			46.4.1 In Vitro Model
			46.4.2 Isolated Organ Model
			46.4.3 In Vivo Model
		Bibliography
	47: Screening Methods for the Evaluation of Anti-Infective Agents
		47.1 Screening of Antifungal Drugs
		47.2 Screening of Anthelminthic Drugs
		47.3 Screening of Antibacterial Drugs
		47.4 Screening of Antiviral Drugs
			47.4.1 HIV-Related Screening Methods
			47.4.2 Non-HIV-Related Screening Methods
		Bibliography
	48: Preclinical Toxicity Studies
		48.1 Introduction
		48.2 Acute Toxicity Study
			48.2.1 Aim of Acute Toxicity Study
			48.2.2 Definition
			48.2.3 Animal Selection
			48.2.4 Number of Animals
			48.2.5 Sex of Animals
			48.2.6 Age of the Animals
			48.2.7 General Care of the Animals
			48.2.8 Administration of the Testing Compound
			48.2.9 Route of Drug Administration
			48.2.10 Selection of Dose Levels
			48.2.11 Observation Period
			48.2.12 Calculation of Medial Lethal Dose 50 (LD50)
				48.2.12.1 Staircase or Up-Down Method of LD50 Calculation
				48.2.12.2 Karber´s Method for Calculation of LD50
				48.2.12.3 Miller and Tainter´s Graphical Method of Calculation of LD50
				48.2.12.4 LD50 Calculation by Lorke´s Method
			48.2.13 Summary of Acute Toxicity Studies
		48.3 Sub-Acute Toxicity Studies (Repeated 14-28Days Toxicity Study)
			48.3.1 Aim
			48.3.2 General Considerations
			48.3.3 Calculation of Sub-Acute Lethal Dose
		48.4 Sub-Chronic (90Days Repeated Dose) Toxicity Study
			48.4.1 Aim
			48.4.2 General Considerations
			48.4.3 Calculation of Sub-Chronic Lethal Dose
		48.5 Chronic (180Days - 1Year Repeated Dose) Toxicity Study
			48.5.1 Aim
			48.5.2 General Considerations
			48.5.3 Calculation of Chronic Lethal Dose
			48.5.4 Calculation of NOAEL and LOAEL
			48.5.5 Calculation of Reference Concentration (RfC) or Reference Dose (Rfd) or Acceptable Daily Intake (ADI) from NOAEL or LOA...
		48.6 Reproductive Toxicity Studies
			48.6.1 General Considerations
			48.6.2 Protocols
			48.6.3 Assessment of Male Reproductive Toxicity
			48.6.4 Assessment of Female Reproductive Toxicity
			48.6.5 Endpoints and Indices in Reproductive Toxicity Studies
		48.7 Local Toxicity Studies
			48.7.1 Dermal Toxicity Studies
			48.7.2 Ocular Toxicity Studies
			48.7.3 Inhalational Toxicity Studies
		Bibliography
	49: Ethical Issues in Animal Research
		49.1 Introduction
		49.2 Terms Related to Research in Animals: The Definitions
		49.3 Animal Experimentations: To Do or Not to Do?
		49.4 The R Principles: 3Rs, 4Rs, and so on
			49.4.1 Replacement: The 1st R
			49.4.2 Reduction Alternative: The 2nd R
			49.4.3 Refinement Alternative: The 3rd R
			49.4.4 Miscellaneous Principles
		49.5 Alternatives to Animal Experimentations
		49.6 Evolution of Ethics in Animal Research
		49.7 Organisation for Economic Co-operation and Development (OECD) Guidelines
		49.8 Compendium of CPCSEA
			Box 49.1: Ethical principles adopted by CPCSEA for use of animals in scientific experiments
		49.9 Guide for the Care and Use of Laboratory Animals, 8th edition
		49.10 Animal Experimentation in India and Elsewhere: What Is the Current Status?
		49.11 Conclusion
		Bibliography
	50: Good Laboratory Practice (GLP)
		50.1 Introduction
		50.2 Principles of Good Laboratory Practice
			50.2.1 Test Facility: Organization and Personnel
				50.2.1.1 Test Facility Management
				50.2.1.2 Study Director
				50.2.1.3 Principal Investigator
				50.2.1.4 Study Personnel
			50.2.2 Quality Assurance Programme (QAP)
			50.2.3 Facilities
			50.2.4 Apparatus, Material, and Reagents
			50.2.5 Test Systems
			50.2.6 Test and Reference Items
			50.2.7 Standard Operating Procedures (SOPs)
			50.2.8 Performance of the Study
			50.2.9 Reporting of Study Results
			50.2.10 Storage and Retention of Records and Materials
		50.3 GLP Advantages and Drawbacks
			50.3.1 Advantages of GLP
			50.3.2 Drawbacks of GLP
		50.4 National Good Laboratory Practice (GLP) Compliance Monitoring Authority (NGCMA)
		Bibliography
Part II: Biochemical Pharmacology
	51: Basic Principles and Applications of Simple Analytical Methods
		51.1 Introduction
		51.2 Qualitative Drug Tests
		51.3 Analytical Techniques
			51.3.1 Titrimetric Techniques
			51.3.2 Spectrophotometry and Beer Lambert´s Law
		51.4 Sample Preparation for Drug Analyses
		Bibliography
	52: Principles of Quantitative Estimation of Drugs, Endogenous Compounds, and Poisons-1
		52.1 Chromatography
		52.2 Types of Chromatography
		52.3 Paper Chromatography
			52.3.1 How It Works
		52.4 Thin-Layer Chromatography (TLC)
			52.4.1 How It Works
		52.5 Gas Chromatography (GC)
			52.5.1 Working Principle
			52.5.2 Mobile Phase
			52.5.3 Stationary Phase
			52.5.4 Samples for GC Analysis
			52.5.5 Instrumentation
			52.5.6 Gas Chromatography: Mass Spectrometry (GC-MS)
		52.6 Liquid Chromatography (LC)
			52.6.1 Modern Liquid Chromatography
			52.6.2 Working Principle
			52.6.3 Instrumentation
				52.6.3.1 Solvent Delivery System
				52.6.3.2 Sample Injectors
				52.6.3.3 Columns
				52.6.3.4 Detectors
			52.6.4 Chromatogram Output
			52.6.5 LC-MS
		52.7 Mass Spectrometry
		52.8 Working Principle and Instrumentation
			52.8.1 Sample Introduction
			52.8.2 Ionization
			52.8.3 Mass Analyzers
			52.8.4 Tandem MS
			52.8.5 Detector
		52.9 Enzyme-Linked Immunosorbent Assays
			52.9.1 Working Principle
			52.9.2 Types of ELISA
				52.9.2.1 Direct ELISA
				52.9.2.2 Indirect ELISA
				52.9.2.3 Sandwich ELISA
				52.9.2.4 Competitive/Inhibition ELISA
			52.9.3 Enzymes Commonly Used in ELISA
			52.9.4 Different Steps Performed in ELISA
		Bibliography
	53: Principles of Quantitative Estimation of Drugs, Endogenous Compounds, and Poisons-2
		53.1 Colorimetry
			53.1.1 Introduction
			53.1.2 Principle
			53.1.3 Instrumentation
			53.1.4 Methodology
			53.1.5 Applications
				53.1.5.1 Applications of Colorimetry in Pharmacology
				53.1.5.2 Advantages of Colorimetry
			53.1.6 Limitations of Colorimetry
		53.2 Spectrophotometry
			53.2.1 Introduction
			53.2.2 Principles of Spectrophotometry
			53.2.3 Instrumentation
			53.2.4 Types of Spectrophotometers
			53.2.5 Sample Measurement
			53.2.6 Applications of Spectrophotometer
			53.2.7 Drawbacks or Disadvantages of Spectrophotometer
			53.2.8 Safety Considerations
		53.3 Photometry
			53.3.1 Introduction
				53.3.1.1 Key Terms in Photometry
			53.3.2 Principle
			53.3.3 Instrumentation
			53.3.4 Methodology
			53.3.5 Applications of Photometry
			53.3.6 Advantages
			53.3.7 Limitations
		53.4 Flame Photometry
			53.4.1 Introduction
			53.4.2 Principle
			53.4.3 Instrumentation
			53.4.4 Working of Flame Photometry
			53.4.5 Methodology
			53.4.6 Applications of Flame Photometry
			53.4.7 Advantages
			53.4.8 Disadvantages
		53.5 Fluorimetry
			53.5.1 Introduction
			53.5.2 Principles of Fluorimetry
			53.5.3 Instrumentation
			53.5.4 Fluorescence Measurement
			53.5.5 Applications of fluorometry
		53.6 Radioimmunoassay (RIA)
			53.6.1 Background
			53.6.2 Introduction
			53.6.3 Principle
			53.6.4 Types of RIA
			53.6.5 Materials Required
			53.6.6 Methodology
			53.6.7 Applications
			53.6.8 Advantages
			53.6.9 Limitations
		53.7 Nuclear Magnetic Resonance (NMR) Spectroscopy
			53.7.1 Introduction
			53.7.2 NMR Principle
			53.7.3 NMR: Sample handling and working
			53.7.4 Instrumentation
			53.7.5 NMR Interpretation
			53.7.6 Applications of NMR Spectroscopy
		Bibliography
	Untitled
	54: Plant Extraction Methods
		54.1 Introduction
		54.2 Classification
		54.3 Traditional Extraction Methods
			54.3.1 Maceration
			54.3.2 Soxhlet Extraction
			54.3.3 Reflux Extraction
			54.3.4 Decoction
			54.3.5 Infusion
			54.3.6 Percolation and Re-percolation
		54.4 Novel/Modern (Non-conventional) Plant Extraction Methods
			54.4.1 Microwave Extraction
			54.4.2 Ultrasound-Assisted Extraction (Sonication Extraction)
			54.4.3 Supercritical Fluid Extraction
			54.4.4 Pressurized Liquid/Solvent Extraction
			54.4.5 Pulsed Electric Field Extraction
			54.4.6 Vibrocavitation Homogenizer Extraction
			54.4.7 Enzyme Assisted Extraction
			54.4.8 Hydro/Steam Distillation
			54.4.9 Two Phase Extraction
		54.5 Conclusion
		Bibliography
Part III: Research Methodology
	55: Literature Search
		55.1 Introduction
			Box 55.1. Various Reasons for Conducting a Literature Search
		55.2 Types of Medical Literature
		55.3 Types of Literature Search
			55.3.1 Electronic Research Databases
		55.4 Methodology of Literature Search
			55.4.1 Translating Research Question to Keywords (Refer Fig. 55.5)
			55.4.2 Searching all the Sources: Keyword Search
			55.4.3 Techniques to Improve Literature Search
			55.4.4 Keeping a Record of Search Activity
			55.4.5 Reviewing and Refining the Search Results by Further Literature Search
		Bibliography
	56: Study Designs
		56.1 Introduction
		56.2 Evidence-Based medicine
		56.3 Concept of Variables
		56.4 Types of Research
		56.5 Importance of Study Designs
		56.6 Types of Study Designs
			56.6.1 Descriptive Studies
			56.6.2 Analytical Studies
			56.6.3 Interventional Studies
		56.7 Miscellaneous Study Designs
		56.8 Levels of Evidence
		Bibliography
	57: Ethical Issues Related to Medical Research on Human Participants
		57.1 Introduction
			57.1.1 Definitions
			57.1.2 Respect for Autonomy
				Box 57.1. Essential Components of Informed Consent Document
			57.1.3 Nonmaleficence
			57.1.4 Beneficence
			57.1.5 Justice
		57.2 History of Medical Research Ethics: How It Started?
			57.2.1 Declaration of Helsinki
			57.2.2 Council for International Organizations of Medical Sciences (CIOMS) Guidelines
			57.2.3 The Common Rule (The Final Rule)
			57.2.4 Indian Context
		57.3 Ethics Review Committees
		57.4 Ethics in Different Contexts
			57.4.1 Underprivileged Populations
			57.4.2 Publication Ethics
		57.5 Medical Research Ethics: What is the Current Status?
		Box 57.2. Placebos in Clinical Research
		57.6 Conclusion
		Bibliography
Part IV: Biostatistics
	58: Basic Principles and Application of Statistics in Drug Research
		58.1 Why to Use Statistics?
		58.2 Uses of Statistics in Drug Research
		58.3 Some Important Terms
			Box 58.1
		58.4 Important Principles for the Application of Statistics in Clinical Research
		Bibliography
	59: Calculation of Basic Statistical Parameters
		59.1 Introduction
		59.2 Variable
			59.2.1 Types of Variables
				59.2.1.1 Quantitative Variable (Numeric Variable)
					Discrete Variable
					Continuous Variable
				59.2.1.2 Qualitative Variable (Categorical Variable)
					Nominal Variable
					Ordinal Variable
		59.3 Measures of Central Tendency
			59.3.1 Mean
			59.3.2 Median
			59.3.3 Mode
		59.4 Measures of Dispersion
			59.4.1 Standard Deviation
				59.4.1.1 Interpretation of Standard Deviation
			59.4.2 Variance
			59.4.3 Range
			59.4.4 Interquartile Range
				59.4.4.1 Interpretation of IQR
			59.4.5 Normal Distribution Curve
				59.4.5.1 Few Characteristics of the Normal Distribution Curve
			59.4.6 Summary Measures for Qualitative or Categorical Data
				59.4.6.1 Ratio
				59.4.6.2 Rate
				59.4.6.3 Proportion
		59.5 Inferential Statistics
			59.5.1 95% Confidence Interval
		Bibliography
	60: Hypothesis Testing
		60.1 Introduction
		60.2 Inferential Statistics and Its Domains
		60.3 Formulating a Research Question and a Hypothesis
		60.4 Null and Alternative Hypothesis
		60.5 Type I and Type II Errors
		60.6 Steps in Hypothesis Testing
			60.6.1 Example of Hypothesis Testing
		60.7 Interpreting the p-Value
			60.7.1 Problems in Interpretation of p-Value
		60.8 Confidence Interval
		60.9 Multiple Hypothesis Testing
		60.10 Bayesian Hypothesis Testing
		60.11 Conclusion
		Bibliography
	61: Parametric Tests
		61.1 Introduction
		61.2 The Basic Assumptions Underlying Parametric Tests are as Follows (Box. 61.1)
			Box 61.1. Basic Assumptions Underlying Parametric Tests
		61.3 General Steps in Performing any of the Parametric Tests
		61.4 Selection of Appropriate Parametric Test
		61.5 Parametric Testing Methods
			61.5.1 T-Test
				61.5.1.1 One Sample t-Test
				61.5.1.2 Independent Sample t-Test/unpaired t-Test/Student´s t Test
				61.5.1.3 The Paired-Samples t-Test
			61.5.2 Analysis of Variance (ANOVA)
				61.5.2.1 One Way ANOVA Test
				61.5.2.2 Two Way ANOVA Test
				61.5.2.3 Repeated Measures ANOVA
			61.5.3 Pearson´s Correlation
			61.5.4 Regression
		61.6 Advantages of Parametric Test Over Non-parametric Test
		61.7 Circumstances Where Parametric Tests Cannot Be Used
		Bibliography
	62: Non-parametric Tests
		62.1 Introduction to Non-parametric Statistics
		62.2 Scope of Non-parametric Tests
		62.3 Steps in Performing Non-parametric Tests
		62.4 Non-parametric Testing Methods
			62.4.1 Sign Test
			62.4.2 Wilcoxon Signed-Rank Test
			62.4.3 Friedman Test
			62.4.4 Goodman Kruskal´s Gamma Test
			62.4.5 Kruskal-Wallis Test
			62.4.6 Mann-Kendall Trend Test
			62.4.7 Mann Whitney U-Test
			62.4.8 Mood´s Median Test
			62.4.9 Macnemar´s Chi-Squared Test
		62.5 Advantages of Non-parametric Tests
		62.6 Disadvantages of Non-parametric Tests
		Bibliography
	63: Common Errors in Using Statistical Tools and Data Presentation
		63.1 Introduction
		63.2 Errors Related to Sample Size
			63.2.1 Manipulation of Confidence Interval (1-α) and Power (1-beta) Values to Achieve Low Sample Size
			63.2.2 Extrapolating Conclusions for an Objective From the Sample Size Estimated for Different Objectives
			63.2.3 Missing Drop-Out Cases in Sample Size Calculation
		63.3 Errors Related to Data Handling
			63.3.1 Unnecessary Conversion of Continuous Data to Categorical Data
			63.3.2 Setting a Groundless Cut-off Value During Categorization
			63.3.3 Unnecessary Data Transformation
		63.4 Errors Related to Statistical Tests
			63.4.1 Choosing Correct Statistical Tests
			63.4.2 Violating the Assumptions of Statistical Test
		63.5 Errors in Data Representation
			63.5.1 Errors in Data Tabulation
			63.5.2 Errors in Graphs
			63.5.3 Errors in Descriptive Data Summarization
				63.5.3.1 Errors in P-Value Expression
		Bibliography
Correction to: Hypothesis Testing
	Correction to: Chapter 60 in: M. Lakshmanan et al. (eds.), Introduction to Basics of Pharmacologyand Toxicology, https://doi.o...