Preclinical Screening of Antiasthmatic DrugsShubham Kolge
Bronchial asthma is characterized by both bronchoconstriction and airway inflammation which leads to bronchial hyperresponsiveness to various stimuli. Different mediators are implicated in asthma. As the precise etiology is not known and multiple biochemical processes are triggered by different causative factors, it is difficult to have a single drug which can effectively and simultaneously act upon different mediators. This led to an intense search for potent and safe antiasthmatic drugs. This presentation intends to compile different screening methods for the evaluation of new candidate drugs with potential for the treatment of asthma. These include in vitro, in vivo, receptor binding and enzymatic methods.
Preclinical Screening of Antiasthmatic DrugsShubham Kolge
Bronchial asthma is characterized by both bronchoconstriction and airway inflammation which leads to bronchial hyperresponsiveness to various stimuli. Different mediators are implicated in asthma. As the precise etiology is not known and multiple biochemical processes are triggered by different causative factors, it is difficult to have a single drug which can effectively and simultaneously act upon different mediators. This led to an intense search for potent and safe antiasthmatic drugs. This presentation intends to compile different screening methods for the evaluation of new candidate drugs with potential for the treatment of asthma. These include in vitro, in vivo, receptor binding and enzymatic methods.
Pharmacological screening of Anti-psychotic agentsAbin Joy
Presentation contents are:
Introduction, Definition of psychosis, Classification of anti-psychotics, MOA of anti-psychotic agents and screening models.
In this slide contains diabetics, classification, symptoms, complication, invivo and invitro screening models of anti diabetics.
Presented by: GEETHANJALI ADAPALA (Department of pharmacology).
RIPER, anantapur
Pharmacological screening of Anti-psychotic agentsAbin Joy
Presentation contents are:
Introduction, Definition of psychosis, Classification of anti-psychotics, MOA of anti-psychotic agents and screening models.
In this slide contains diabetics, classification, symptoms, complication, invivo and invitro screening models of anti diabetics.
Presented by: GEETHANJALI ADAPALA (Department of pharmacology).
RIPER, anantapur
ISSN 2347-2251
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The Indo-American Journal of Pharma and Bio Sciences is an online international journal that publishes articles quarterly.It's important to note that the specific policies, guidelines, and the editorial board of IAJPB may change over time, so it's advisable to visit the journal's official website or contact the journal of the research on journaling.
ABSTRACT
Background:The main objective of the study is to determine the anti-arthritic effect of whole plant ethanolic extract of Polygonum glabrum
belonging to the family Polygonaceae in Female wistar rats using the Freund’s Complete Adjuvant (FCA) model . Methods:The plants areal
parts were collected near Tirupathi hills, Chittoor district of Andhra Pradesh in India. The Phytoconstituents were identified through the
chemical tests. Ethanol (95%) was used to obtain the whole plant extraction through Soxhlet extractor. Female SD rats were used for antiarthritic
screening. Arthritis was induced using FCA, and the anti-arthritic effect of the ethanolic extract of P.glabrum was studied at doses
of 250 and500 mg/kg. The effects were compared with those of indomethacin (10 mg/kg). At the end of the study, theliver enzyme levels were
determined and a radiological examination was carried out. Results and Discussion:The preliminary phytochemical analysis of the ethanolic
extract of Polygonum glabrum showed the presence of alkaloids, tannins, flavonoids and saponins. P. glabrum at 250 and 500 mg/kg
significantly inhibited the FCA-induced arthritis in the rats. This was manifested by as a decrease in the paw volume. The arthritic control
animals exhibited a significant decrease in body weight compared with control animals without arthritis. P. glabrum treated animals showed
dose dependent reduction in decrease in body weight and arthritis.At the same time, P.glabrum significantly altered the biochemical and
haematological changes induced by FCA (P < 0.05). The anti-arthritic effect of P.glabrum was comparable with that of Indomethacin.
Conclusion:The whole plant extract of P.glabrum showed significant anti-arthritic activity against FCA-induced arthritis in female Wistar
rats.
Intercontinental journal of pharmaceutical Investigations and ResearchSriramNagarajan19
Anti-inflammatory activity of the ethanolic extract of Portulaca quadrifida Linn. was studied in wister rats using the carrageenan induced left hind paw edema, carrageenan induced pleurisy and cotton pellet induced granuloma model. The ethanolic extract (200 mg/kg, p.o.,) produced the inhibition of carrageenan induced rat paw edema. It also showed an inhibitory effect on leukocyte migration and a reduction on the pleural exudates as well as reduction on the granuloma weight in the cotton pellet granuloma method. The results indicated that the ethanolic extract produced significant (P<0.001) anti-inflammatory activity when compared with the standard and untreated control.
Enterocin 55 produced by non rabbit-derived strain Enterococcus faecium EF55 ...Agriculture Journal IJOEAR
— Ent55 is produced by poultry strain Enterococcus faecium EF55. It is substance which can be allotted to Class II enterocins; thermo-stable, small peptide. Because producer strain has shown beneficial effect in poultry and broiler rabbits as well, we decided to apply Ent55 in broiler rabbit husbandry. Ent55 showed antimicrobial activity in broiler rabbits by reduction of staphylococci, Clostridiae, pseudomonads and coliforms. Its beneficial effect was demonstrated by stimulation of phagocytic activity as well as by reduction of Eimeria spp. oocysts. GPx values were lower; it means, no oxidative stress was evoked. Moreover, it has not negative influence on growth performance and biochemical parameters. Our results indicated that enterocin produced by not-autochtonous strain can also have protective and beneficial effect in broiler rabbits.
In-Vivo Evaluation of Rifampicin Loaded Nanospheres: Biodistribution and Myco...Ratnakaram Venkata Nadh
Rifampicin PLGA nanospheres are
formulated with a specific goal in order to decrease
the dose, adverse effects and to enhance targeted
drug delivery. Rifampicin nanospheres were
prepared and evaluated by emulsion solvent
evaporation method. In vivo bio distribution studies
reveal that there was a long term accumulation of
rifampicin nanospheres in the lungs over other
organs. The increase in Cmax values confirmed that
inhalable PLGA nanospheres are suitable for
targeting and providing sustained release of antitubercular
drugs to lungs. So inhalation is a
selected administration route of Rifampicin PLGA
nanospheres. The in vivo screening of M.
tuberculosis showed good activity as well as its
activity against multidrug-resistant M. tuberculosis
and against M. tuberculosis isolates in a
potentially latent state, makes Rifampicin PLGA
nanospheres as an attractive drug dosage form
for the therapy of tuberculosis. It can be concluded
that there is a significant potential for effective
oral delivery as well as nasal delivery of the
Nanospheres for the treatment of tuberculosis.
Obesity is very serious and concerned problem these days. Despite availability of many drugs in market to treat
obesity, no single drug is ideal for treating all sorts of problems caused by obesity. The obesity models available
for inducing obesity are by using chemicals and high fat diet. Wistar albino rats were used to study anti-obesity
activity of methanolic extract of Tricholepisglaberrima plant aerial parts at doses 100 mg/kg p.o. and 200
mg/kg p.o. against the standard orlistat 50 mg/kg p.o. in models of anti-obesity activity viz. High fat induced
obesity, Monosodium glutamate induced obesity model. The induction of obesity is done by diet (20
grams/animal/day) and Monosodium glutamate (oral). The study period is 28 days for both models. In both
models, the plant showed anti-obesity activity significantly at a dose of 100mg/kg and 200 mg/kgp.o. by
reducing the body weight, fat pads weight, total cholesterol, triglycerides, LDL, VLDL, biomarkers enzymes like
SGOT, SGPT and ALP, whereas significant increase in HDL levels was observed. Further multiple dose preclinical studies and clinical studies have to be carried out for proving for human obesity treatment.
The biological activities of methanolic extract of
Tricholepisglaberrima observed in this study
strongly indicated their great potential as anti-obese
and obesity associated complications like
hyprlipidemia. Oral administration of extracts
reduced the level of circulating lipids significantly,
resulting in the decrease of body weights in various
animal models of obesity bearing close
resemblance to human obesity. Extract appear to
show such activities by modulating the lipid
metabolism through the decreased activity in
lipogenesis or by inhibition of pancreatic lipase
activity.
The methanolic extract of aerial parts of
Tricholepisglaberrima at a dose of 200mg/kg b.w.
p.o. significantly reduced total cholesterol,
triglycerides, LDL, VLDL, biomarkers enzymes
like SGOT, SGPT and ALP, whereas significant
increase in HDL levels was observed.
Phytoconstituents like saponins, tannins and
flavonoids in METG may be responsible for its anti-obesity and anti-hyperlipidemic activities by
multiple actions.
Apart from anti-obesity and anti-hyperlipidemic
agent, It may also act as hepatoprotective agent due
to possessing significant reduction in SGOT, SGPT
and ALP levels and significant increase in HDL
levels respectively.
Thus it can be said that METG is effective in
ameliorating abnormalities in lipid profile and fat
accumulation in rats and results provides useful
information for the clinical research that this plant
can be used as herbal drug in the treatment of
obesity and hyperlipidemia. Further studies on this
extract may be focused on the possible mechanism
of action, isolation, characterization and
purification of active constituents which is
responsible for anti-obesity and anti-hyperlipidemic
activities.
EFFECT OF DIFFERENT CHROMATOGRAPHIC FRACTION AQUEOUS AND ALCOHOLIC EXTRACTS O...Jing Zang
In recent studies Teucrium polium(T. polium ) was known as a hypoglycemic plants. But further research is needed to better understand the effect of Teucrium polium and biological active part of it. The purpose of this investigation is to examine the effect of different chromatographic fractions of aqueous and alcoholic extract of this plant on the level of insulin secretion and glucose content in hyperglycemic rat model. Also, our aim is determination of biological active fraction of aqueous and alcoholic extract of this plant. This study was carried out on the 36 rats. Hyperglycemia induced by administrating of 50 mg/kg alloxan intraperitoneally and glucose level was monitored for hyperglycemic status. Hyperglycemic was confirmed by blood glucose measurement. In each experiment 100 grams of Teucrium polium aerial parts powder were boiled with 2 Litter of distilled water for 36 h. The decoction preparation was then filtered through a gauz cloth followed by filtration through filter paper. The extract was evaporated to one-fifth of its original volume and kept at 4oC until its use. Determination of different fraction aqueous extract effect of Teucrium polium on glucose level and insulin secretion was carried out. Blood was collected from the tail of the rats. Then glucose and insulin level was evaluated. The hyperglycemic animals showed significant decrease in the blood glucose level in rats administered with fourth fraction compared with other factions. Administration of fourth fraction Teucrium polium aerial parts extract cause increase in insulin levels in alloxan-treated rats. Results suggest that treatment of fourth fraction Teucrium polium aerial parts extract may be useful in preventing the increase of glucose level in hyperglycemic rats. The interesting phenomenon of our results has shown that fourth fraction given parenterally possesses a hypoglycemic effect in alloxan hyperglycemic rats. Fourth fraction was found biological active and to be responsive to glucose challenge as evidenced by increase in insulin secretion.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Similar to Evaluation methods of anti-asthmatics (20)
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
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Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
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
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
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Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
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Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
Maxilla, Mandible & Hyoid Bone & Clinical Correlations by Dr. RIG.pptx
Evaluation methods of anti-asthmatics
1. Dr Manjeeta Gupta
Evaluation of anti-asthmatic drugs
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 1
2. Asthma
Global health problem resulting from complex
interaction between genetic & environmental
factors
Nearly 7–10% of world population suffers from
bronchial asthma
Among several respiratory diseases, bronchial
asthma is most common disabling syndrome
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 2
3. 7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 3
Chronic, heterogeous,
inflammatory disorder of
airways
characterized by:
1. Airway obstruction
2. Airway inflammation
3. Bronchial hyper-
responsiveness
In this screening method, we measure potency of anti-asthmatic agents,
by inducing bronchial hyperactivity in experimental animals (rats, mice, guinea pigs, monkeys)
4. Acute toxicity test
Before starting any in vivo assays, it is important to study oral acute
toxicity, for selection of test dose
Initial dose finding procedure Albino mice (either sex, 20-25g)
Group 1 – 3 mice given 10mg/kg test drug i.p.
Group 2 – 3 mice given 100mg/kg test drug i.p.
Group 3 – 3 mice given 1000mg/kg test drug i.p.
(Monitor for 24 hrs)
From above results we take 4 doses & administer i.p.
4 groups 1 mouse/group
LD50 Mean of lowest dose showing death & highest dose not showing
death
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 4
5. Ideal animal models for asthma
Similar genetic basis to human diseases
Similar anatomy & physiology
Similar pathological response
Respond to drugs with known clinical efficacy
Predict clinical efficacy
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 5
6. In vitro methods
1. Binding Assay
a. Histamine receptor assay
2. Cell culture methods
a. CULTEX technique
b. WST assay
3. Tests in isolated organs
a. Spasmolytic activity in guinea pig lungs
b. Vascular & airway responses to isolated lung
c. Reactivity of isolated perfused guinea pig trachea
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 6
7. In vivo methods
1. Bronchospasmolytic activity in anaesthetized guinea pigs
2. Arachidonic acid/PAF induced respiratory vascular dysfunction
3. Anaphylactic microshock
4. Serotonin aerosol induced asphyxia
5. Histamine induced bronchoconstriction
6. Pneumatochography in guinea pigs
7. Bronchial hyperactivity in guinea pigs
8. Mast cell stabilising activity on rat mesentery
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 7
9. Histamine receptor assay
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 9
Animal used Guinea pig brain plasma (Male, 300-600g)
Aim To measure inhibitory activity of test compound on
binding of 3H pyrilamine (H1 antagonist)
1. Total binding
2. Non specific binding
3. Specific binding (Total binding – Non specific binding)
4. % inhibition of 3H pyrilamine binding (100-specific
binding)
11. CULTEX technique
New experimental method for cultivation & exposure of cells of
respiratory tract to air borne pollutants at air/liquid interface
Enhanced efficiency of in-vitro studies
Principle Direct exposure of bronchial epithelial cells to
complex mixtures
Aim To study factors influencing susceptibility of human bronchial
epithelial cells
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 11
12. 7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 12
Bronchial epithelial cells
washed with PBS
Incubate with test drug for
24hrs
Cells exposed to clean
air/different concentrations
of smoke for 1 hr (Cell
exposure unit)
Procedure
13. WST assay
Transfer cells from cell
exposure vessel to
plates containing 2ml
fresh RPMI medium
Add 500μl RPMI &
WST-1 dye & incubate
for 1hr
Aliquots transferred
into 96 well microplate
(for absorbance at 450-
630 nm)
Measurement of
absorbance by
microplate reader
Cells are trypsinized
by adding 500μl
trypsin/ETDA soluton
Incubate at 37˚C for 4
mins
Add 25μl trypsin
inhibitor
Gently suspend cells &
dilute 100μl
suspension in 9.9ml
CASYton solution
Analyse aliquots with
Electronic cell counter
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 13
14. Spasmolytic activity in lungs
Animal used Albino guinea pig (either sex, 300-450g)
Preparation Animal sacrificed with overdose of ether
Aim To evaluate capability of inhibiting bronchospasm induced by histamine
& Calcium ionophores
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 14
16. Vascular & airway responses to isolated lung
Animal used Sprague dawley rats (300-350g)
Anaesthetic used Pentobarbitone sodium (50mg/kg) i.p
Aim To measure & compare..
1. Pulmonary arterial pressure
2. Airway pressure
3. Reservoir blood level
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 16
17. Trachea cannulated &
animal on artificial
respiration
Rat heparinized with
1000 IU & blood
withdrawn from carotid
artery
Lungs removed (median
sternotomy) &
suspended water
jacketed chamber
Pulmonary artery
catheterised
Lungs perfused with
Krebs-Henseleit
solution
Changes in parameters
recorded after addition
of test drug &
compared with baseline
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 17
Procedure
18. Reactivity of isolated perfused trachea
Animal used Albino guinea pig (either sex, 300-550g)
Preparation Sacrificed by CO2 narcosis
Aims
1. To study mechanism by which epithelium affects
reactivity of tracheal musculature
2. To study effects of histamine, Calcium ionophores,
bradykinin, leukotriene, potassium channel openers on
tracheal musculature
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 18
19. Trachea dissected & cut into individual rings
12-15 rings tied together & mounted in organ bath
(Krebs-Henseleit buffer solution)
Tissue maintained at 37˚C & bubbled with carbogen
Catheters (inlet-outlet) connected to positive & negative sides of polygraph
Response of tracheal musculature by changing inlet-outlet pressure is recorded
After 45 mins spasmogens added
When maximum contraction is reached standard drug is added (isoprenaline/aminophylline)
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 19
Procedure
20. Readings recorded
Tissue washed
Controlled contractions induced again by addition of spasmogens
Record contractions & add test drug
Record readings
(Change in pressure in cm of water is taken as response)
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 20
22. Bronchospasmolytic activity (Konzett-Rosseler method)
Animal used Guinea pig (250-500g)
Anaesthetic used 1.25g/kg urethane i.p.
Principle Bronchospasm causes ↓ volume of inspired air & ↑
volume of excess air
Aim To measure volume of air not taken up by lungs after
bronchospasm
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 22
23. Trachea is cannulated
ARM 1 connected to respiratory pump
ARM 2 connected to statham pressure tranducer
Artificial ventilation at frequency 60strokes/min is maintained
Excess air not taken up by lungs is measured
Test drug administered (through jugular vein)
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 23
Procedure
24. BP recorded (from carotid artery)
Each animal placed in plastic containers (Histamine chamber) of 15L volume
0.25% histamine solution aerosol sprayed at 180mmHg
5 mins exposure time (Test drug given orally 1 hr before exposure)
Spasmogen challenge is repeated
Unprotected animals fall on their sides
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 24
25. Arachidonic acid/PAF induced respiratory & vascular
dysfunction
Animal used Guinea pigs (Male, 300-600g)
Anaesthetic used Pentobarbitone sodium 60mg/kg i.p.
Principle Thromboxane causes bronchoconstriction & thrombocytopenia
Prostacyclin cause ↓ SBP & ↓ DBP
Aim To study & compare…
1. % inhibition or increase of bronchospasm
2. BP reduction (measure magnitude & duration)
3. Thrombocytopenia & haematocrit
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26. Procedure
Trachea artificial respiration (70-75 strokes/min)
Jugular vein test drug
Carotid artery blood withdrawal & transducer attached for BP
measurement
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Record BP & changes in airflow
Animal given multiple intravenous injections of arachidonic
acid till 2 bronchospasms of equal intensity are obtained
Test drug administered intravenously
Repeat spasmogen injections
27. Anaphylactic Microshock
Animals used Guinea pig (200-300g) sensitized with s.c injection of
egg albumin
Rabbit (2000-3000g) sensitized with 2% histamine
aerosol
Ova transgenic mouse model (mouse sensitized with
ova i.p Gold standard model
Aim Measure degree of protection (p) = [1- (C/T)] * 100
Where, C = control animals
T = treated animals
(C ≤ 40 secs ≥ 165 secs are excluded)
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 27
29. Serotonin aerosol induced asphyxia
Animal used Guinea pig (200-300g)
Principle Serotonin causes bronchoconstriction asphyxia & death
Aim To evaluate % protection = (1- T1/T2)* 100
where, T1 = mean of control pre-convulsion time 2 days
before & 2 days after administration of drug
T2 = Pre-convulsion time with administration of drug
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31. Histamine induced bronchocostriction
Animal used Guinea pig (400-600g)
Anaesthetic used Pentobarbitone 70mg/kg i.p.
Prniciple Bronchodilators attenuate ↓ respiratory amplitude
& ↑ respiratory frequency after histamine inhalation
Aims To study & calculate…
1. Respiratory frequency
2. Respiratory amplitude
3. Time required for antagonism against bradykinin induced
bronchoconstriction
4. Bronchodilator effects of Potassium channel openers
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32. Procedure
Trachea artificial respiration (60 strokes/min)
Jugular vein test drug
Carotid artery blood withdrawal & transducer attached for BP
measurement
Other parameters:
Airflow rate Differential pressure transducer
Tidal volume & trans pulmonary pressure
Pulmonary resistance (PR) & dynamic lung compliance (LC)
Systemic BP Statham pressure transducer
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33. 7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 33
Histamine intravenous
injection (↓LC & ↑PR by
200%)
Repeat after 5mins
After 3 reproducible
responses
Test drug given
intravenously 1min before
histamine injection
Inhibition of histamine
induced bronchoconstriction
recorded
34. Pneumatography in guinea pigs
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 34
Animal used Guinea pig (300-400g)
Anaesthetic used Urethane (1.5g/kg, i.p.)
Aim To measure respiratory & circulatory parameters
Procedure Trachea is cannulated (connected to pnematograph)
Catheter placed in oesophagus (Oesophageal transducer)
with tip inside thorax (registers intrathoracic pressure)
Cephalic vein & carotid artery cannulated (Gould
pressure transducer)
Test & control readings recorded
35. Mast cell stabilising activity on rat mesentery
(Kaley & Weiner)
Animal used Male albino rats
Aim To study % of degranulated mast cells
Procedure
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 35
Small intestine with mesentery (Petri dish with PBS)
Mesentery incubated with different concentrations of disodium
cromoglycate
Challenged with 1μg/ml 48/80 (standard granulator) for 10mins
Mesentery stained with 0.1% toluidine blue for 20-30mins
Mount pieces on slide
36. Non human primate models
Animals used Rhesus monkeys
Cynomologus monkey
Spasmogens House dust mite, ascaris, pollen antigens
Aims To study…
1. Early & late phase bronchoconstriction response
2. Airway eosinophilia
3. Acute hyperactivity response
4. Human proteins & monoclonal antibody therapeutics
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37. Alternatives to animal models
Cell culture
Patient sputum culture
BAL fluid sampling
Direct reprogramming of patient fibroblasts (patient derived
induced pluripotent stem cells)
Newer in vitro techiques – Human tissue explants
Precision cut lung slices
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 37
38. References
Drug screening methods by S.K. Gupta
Animal models of asthma: reprise or reboot? Biochemical pharmacology
Evaluation of antiasthmatic activity of a polyherbal formulation containing four
plant extracts. Journal of current pharmaceutical research
Measuring the lung function in the mouse: the challenge of size. Respiratory
research
Various screening methods of anti-allergic activity. International journal of
pharmaceutical sciences and nanotechnology
Vogel
7/8/2016Department of Pharmacology,MIMER Medical College Talegaon Dabhade 38
Editor's Notes
Tris solution incubation buffer to determine total binding
Non specific binding determined in d presence of mepyramine (same as pyrilamine)
Calculate % inhibition
Biological parameters studied
Number of cells
Metabolic activity
Glutathione concentration
Cell viability measurements
CULTEX technique helps to use samples for subsequent in vitro assays.
WST- water soluble tetrazolium salts (cell proliferation reagent)
RPMI medium Roswell Park Memorial Institute medium. Used for cell/tissue culture
CASYton solution ready to use isotonic & isosmotic dilution liquid for cell culture
Measures cell viability in test & control group along with biological parameters
Contractile dose is determined isometrically
Calcium causes activation of leukotrienes via 5-lipoxygenase
To test lungs ability to contract
Spasmoges carbachol, histamine diHCl, Ca ionophores, leukotrienes C4 & D4
Preload readings for maximal contractions
Recorded by polygraph
Thromboxane & prostacyclin are byproducts of arachidonic acid metabolism
Compare test & control (before treatment)
Compare results before & after drug administration
Microshock one that is interrupted before death & is repeatable
Pre-convulsion time time from commencement of exposure to severe dyspnoea is recorded. Measures severity of shock
1 & 2 measured by plethysmograph
Guinea pig most sensitive to histamine
Ringer locke physiological solution
Anatomical & structural similarity between humans & monkeys
Sensitization develops in 18 months
Airway function & BAL fluid sampling