This document provides information on various isolated tissue preparations that can be used to study the effects of drugs. It discusses the advantages of different in vitro, ex vivo, and in vivo preparations using tissues from animals like guinea pigs, rabbits, rats, mice, cats, and dogs. Specific tissues that are mentioned include isolated strips of intestine, trachea, atria, fundus, vas deferens, anococcygeus, and more. The document outlines the process for setting up these preparations in an organ bath, applying drugs, and measuring responses. It also lists the typical drugs that different tissues are sensitive to and the receptors involved.
Selection of an animal model is one of the most important steps in any of the experimental pharmacological study.
Animal model preferred for the study must be producing similar disease profile as in the human.
Expt. 6 Bioassay of histamine using guinea pig ileum by matching methodVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of histamine standard solution
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
Selection of an animal model is one of the most important steps in any of the experimental pharmacological study.
Animal model preferred for the study must be producing similar disease profile as in the human.
Expt. 6 Bioassay of histamine using guinea pig ileum by matching methodVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of histamine standard solution
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
The presentation is about the dose selection for laboratory animal toxicology drug testing, explaining staged and staggered approach of dose selection.
Expt. 4 DRC of acetylcholine using frog rectus abdominis muscleVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation of magnification value (Mf)
Graphical presentation of CRC/ DRC
Result and interpretation
Introduction to pre clinical screening of drugsKanthlal SK
Various Techniques and Methods for screening of new chemical entities in preclinical aspects (both invitro & invivo) for effective and safe clinical usage.
Presentation on all the evaluation methods in animals for anti diabetics. It includes methods for insulin dependant and insulin independent diabetes mellitus!
Common laboratory animals, Classification of Experimental Animals, Handling and application of different species and strains of animals,Different strains of laboratory animals, application and common diseases.
The presentation is about the dose selection for laboratory animal toxicology drug testing, explaining staged and staggered approach of dose selection.
Expt. 4 DRC of acetylcholine using frog rectus abdominis muscleVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation of magnification value (Mf)
Graphical presentation of CRC/ DRC
Result and interpretation
Introduction to pre clinical screening of drugsKanthlal SK
Various Techniques and Methods for screening of new chemical entities in preclinical aspects (both invitro & invivo) for effective and safe clinical usage.
Presentation on all the evaluation methods in animals for anti diabetics. It includes methods for insulin dependant and insulin independent diabetes mellitus!
Common laboratory animals, Classification of Experimental Animals, Handling and application of different species and strains of animals,Different strains of laboratory animals, application and common diseases.
Experiment 1 Introduction to In-Vitro pharmacology and physiological salt so...Kanchan Chouksey
In Vitro pharmacology studies the biological effects of a drug in an isolated environment, such as cell lines or tissues.
This setup conveniently eliminates whole organism physiological influences allowing for a detailed analysis a compound’s impact.
Heart is supplied by autonomic nervous system.
Adrenaline acts as agonist.
It acts on β-receptors and increases heart rate and amplitude.
Acetylcholine acts on muscarinic receptors as an agonist and decreases the heart rate and amplitude.
Excess concentration of KCl stops the heart beat during diastolic phase.
Ca2+ excess concentration stops the heart beat during systolic phase.
K+ and Ca2+act on cardiac muscle through non-receptor mechanism of action.
- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
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Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
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Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Rasamanikya is a excellent preparation in the field of Rasashastra, it is used in various Kushtha Roga, Shwasa, Vicharchika, Bhagandara, Vatarakta, and Phiranga Roga. In this article Preparation& Comparative analytical profile for both Formulationon i.e Rasamanikya prepared by Kushmanda swarasa & Churnodhaka Shodita Haratala. The study aims to provide insights into the comparative efficacy and analytical aspects of these formulations for enhanced therapeutic outcomes.
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5. Isolated strips of intestine
Abundant material , more resistant &
easy to set up
Variable spontaneous activity of
different sections
Different type of pharmacological
actions can be studied
7. Process
• Complete adjustment of organ bath
• Adjustment of lever
• Animal and tissue selection
• Surgical process and collection of a tissue
• Tissue attachment to organ bath
• Relaxation time to tissue
8. • Prepare the standard drug
• Test any concentration of drug for response
• Standardize tissue response with same
dose
• Prepare DRC for standard drug
• Prepare DRC with test drug
9. Important Points to Remember
• Check Organ bath and PSS
• Writing Lever
• Dissection
• Rest
• Empty organ bath regularly
• Add drug slowly
10. • Allow drug to act
• Wash
• Put a mark on drum
• Use stopwatch during experiment
• Start with a very low concentration
11. Physiological Salt Solution
• Also called as PSS / Ringer solution
• Maintains tissue outside the animal body
• Select PSS in which tissue last longer
• Prepare the solution with the help of distilled
or double distilled or deionised water
• Prepare fresh solution
13. • McEwen solution - sucrose in addition to
glucose
• Krebs solution - used for any tissue
• DeJalon , Frog-Ringer and Ringer-Locke do not
contain magnesiun and phosphate
• Krebs, DeJalon and McEwen – aerated with
carbogen and used for the mammalian
isolated organ and avian skeletal muscle
14. Fast Contracting Smooth Muscle
Preparation
TISSUE DRUG
Guinea pig ileum Histamine
Rat colon Acetylcholine (ACh)
Rat uterus ACh
Rat anococcygeus ACh
Rat vas deferens ACh
Guinea pig atria Adrenaline (Adr)
15. Guinea Pig Ileum
• AIM :- To determine unknown concentration
of histamine by using guinea pig ileum
• Guinea pig ileum is most sensitive to
histamine .
• It has the spontaneous activity and specificity
is improved by using atropine (for histamine
assay) or mepyramine (for acetylcholine) in
Tyrode.
16. • Ileum is preferred because of less mesentry
attached to it, and nearly all receptors are
present.
• 10 cm of ileum attached to the cecum should
be excluded.
• Spontaneous activity of the tissue is reduced
by performing the experiment 5-7°C lower
than body temperature.
17. Rat Colon
• AIM:- To determine unknown concentration of
ACh using rat ascending/descending colon.
• Sensitivity may increase by keeping colon at
4°C for 24 hr.
• Expression of calcium-sensing receptors(CaSR)
mediates increase in inositol 1,4,5-
trisphosphate.
18. • Easy to isolate and more handling resistant.
• For ACh , most sensitive tissue is dorsal leech
muscle and frog rectus abdominis.
• Initial few centimeters of colon usually used
for bioassay of Adr and NA.
19. Guinea Pig Atria
• AIM:- To determine unknown concentration of
Adr using guinea pig atria
• Beta 2 receptors widely present in atria
• Required less trimming or slicing of the tissue
• Advantages :- Tissue thickness , easy
separation of right and left atria , good
contractile tension , stability etc
20. Rat Uterus
• AIM :-To determine unknown concentration of
ACh using rat uterus
• Induce the estrus cycle in female rat
• Treat with stilbestrol 0.1mg/kg, SC ,
24 hrs before the experiment
• Other drugs like adrenaline , histamine,
oxytocin also sensitive to uterus
21. • Response depends on the animal age due to
variation in estrus cycle .
• Other drugs like 5-HT,NA can be assayed .
22. Rat Vas Deferens
• AIM :- To determine unknown concentration
of ACh using rat vas deferens
• Described by the Henderson et al(1972) and
Hart et al(1979)
• Rat and guinea pig are suitable animals
• Animal should be fed with oats for 3 days .
23. • Adrenaline and phenylephrine contract the
vas deferens whose action is blocked by the
phentolamine .
24. Rat Anococcygeus
• AIM: To determine unknown concentration of
ACh using rat anococcygeus muscle
preparation
• Decribed by Gillespie (1972)
• It arises from sacral vertebrae and reaches to
terminal colon (near anus) .
• It has tendinous origin and do not appear soft.
25. • It has a dense adrenergic excitatory and
inhibitory innervations .
• Insenistive to histamine .
26. Slow Contracting Muscle
TISSUE DRUG
Rat fundus Seretonin (5-HT)
Frog rectus abdominis ACh
Guinea pig trachea ACh
Rat phrenic nerve diaphragm ACh
Chick biventral cervicis Neuromuscular blocking agents
27. Rat Fundus
• AIM :-To determine unknown concentration of
5-HT using rat stomach(fundus)
• Stomach fundus is most sensitive tissue
among the whole parts of stomach.
• fundus identification – gray color , above the
pink thick pylorus.
• Insensitive to histamine
28. • Animal should be fasted properly.
• Fundus contains the swallowed air.
• Sometimes it needs a stretching weight.
• Both longitudinal and circular muscles may be
used in the experiment which depends on the
transverse cut made to prepare the tissue.
29. Frog Rectus Abdominis
• AIM :- To determine unknown concentration
of ACh using frog rectus abdominis muscle.
• It’s a striated skeletal muscle sensitive to Ach
and the easiest isolated tissue to handle.
• Contains more of multiple-innervated fibers so
show the slow contraction.
• Frog, being an amphibian animal, it responds
under room temperature.
30. Guinea Pig Trachea
• AIM :- to determine unknown concentration
of ACh using guinea pig trachea
• Extraction of at least 6 cm of trachea required
• Tracheal ring is in “D” form and smooth
muscle is present in the straight line of “D”
shape.
• Response is slow to develop but last for longer
period
31.
32. • Mainly to demonstrate the respiratory
dominant adrenoreceptor so utilized to study
the bronchodilators like Adr , theophylline.
• Ideal model to study the contractile drugs like
Ach,5-HT and histamine, and their antagonism
can be studied by drugs like Adr ,
aminophylline.
33. Rat Phrenic Nerve Diaphragm
• AIM :- To determine unknown concentration
of ACh using rat phrenic nerve diaphragm
• This is primary motor nerve of the diaphragm
which arises mainly from the 4th cervical
nerve.
• Useful for evaluation for neuromuscular
function
34. Chick Biventer-Cervicis
• AIM : - To determine neuromuscular blocking
drugs using innervated biventer-cervicis
preparation
• Biventer-cervicis – an anatomically complex
tendinous muscle
• Responds differently twitch / contraction to
different stimuli
35. Cardiac Muscle Preparation
Effect of various drugs on Isolated heart
(Langendorff’s preparation)
Effect of different drugs on Normal and
Hypodynamic Rabbit heart
Effect of inotropic and chronotropic effects of
various drugs on Frogs heart
36. Langendorff’s Preparation
• AIM :- To observe the effect of various drugs
on Isolated heart
• In 1897 , Oscar Langendorff established the
isolated mammalian heart preparation.
• Based on principle of retrograde flow in aorta
either at constant flow or constant pressure .
37. • Animals : -
-Albino Rats
-New Zealand Rabbits
-Guinea pig
• 23° ± 2° C temperature , 12:12 hr light and
dark cycle ,free access to tap water and food
ad libitum
38. • Animal is pretreated with the heparin .
• The recording is done by attaching the thread
to a strain gauge transducer , an attachment
with the student physiograph
39.
40.
41. Rabbit Heart
• AIM : To determine effect of different drugs
on Normal and Hypodynamic Rabbit heart
• Hypodynamic heart – defined as the heart
exhibiting subnormal power or force than the
normal one.
• Experimentally ,it is developed by the supply
of the 1/4th of Calcium chloride(CaCl2) than
the required one which reduces the heart
rate.
42. • For normal heart – McEwens solution
• For hypodynamic heart – 1/4th of CaCl2 in
McEwens solution
43.
44. Frog Heart
• AIM :- To demonstrate the Effect of inotropic
and chronotropic effects of various drugs on
frog heart
• The heart is the most common site for the
drug target.
• Positive inotropic & chronotropic drugs like
Adr and NA .
• Negative inotropic & chronotropic drugs like
ACh .
45. Tissues/muscle Identification point Drug sensitivity Receptors
Tracheal chain Above esophagus NA ,A & Ach β2 & M2/M3
Stomach fundus Upper grey part
attached to thick red
Pylorus
5-HT > Ach >
Histamine >
Bradykinin
5HT-D
Ileum Cecum at middle part
& large intestine at
distal part
Histamine >Ach H1, Muscarinic
Ascending colon 4-7 cm from
ileocecum junction
NA>A
Ach,5-HT
β3 , 5HT-2A , 5HT4
Descending
colon
5-7cm above rectum Ach Muscarinic
Anococcygeus Thin muscle strip
arises from the sacral
vertebrae
NA, Ach, 5-HT , isoP,
No histamine
Adrenergic supply,
NANC
Vas deferens Attached to
epididymis
NA & Ach α1, Muscarinic
Uterus Clear two horn above
rectum connected to
ovary
Oxytocin,5-HT,A,NA
Ach
β , α
M2/M3
46. Rabbit Perfused Ear Artery
• Lop-eared rabbit is used
• Central artery is identified , cannulated ,
dissected & perfused .
• Used for bioassay of catecholamines
47.
48. Rabbit Aortic Strips
• Descending thoracic aorta is used .
• Aorta is cut in a close spiral .
• 2 to 4 cms of strips are used for experiment .
• Has a very slight spontaneous tone .
• Shows no rhythmic contraction .
• Responds to Adr and NA.
49. Tissue / Organ Response Receptor
Rat isolated vas deferens and
anococcygeus
Rabbit and guinea pig isolated aorta
Rabbit isolated jejunum
Guinea pig isolated ileum
Contraction
Contraction
Inhibition
Inhibition
Postsynaptic α₁
Rabbit isolated ear artery and jejunum Inhibition Presynaptic α₂
Rabbit isolated perfused heart
Rabbit isolated jejunum
Contraction
Inhibition
β₁
Guinea pig isolated trachea
Rat isolated uterus
Inhibition
Inhibition
β₂
Mouse isolated spleen contraction
Relaxation
Both