RECENT TECHNIQUES USED IN PRECLINICAL PHARMACOLOGY

Presented by Guide

Mr. Gogawale Vinayak G.M.Pharm. (Semester-I )

(Department of Pharmacology)

Dr. U.M.Aswar(Pharmacology)

SINHGAD INSTITUTE OF PHARMACY, NARHEMay 3, 2023

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• Introduction.

• Types of techniques.

• References.

CONTENTS

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For interpretation of the result of the drug and identification of the tumors the several techniques are used in different categories like behavioral study, biochemistry, by doing surgery and imaging techniques.

Recent techniques for preclinical pharmacology

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Behavioral

Biochemistry

Imaging

Surgery

Techniques used in preclinical pharmacology

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Telemetry

Plethysmography

Behavioral

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It is the wireless transmission and reception of measured quantities for the purpose of remotely monitoring.

Implantable telemetry: The implantable telemetry is the device which is implant in animal body.

Telemetry

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• Data collection is possible when animal is conscious.

• Long term recording and monitoring in free environment.

• 24/7 access of data possible.• Reclean , reuse, re-sterilization is not required.

Advantages

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• Monitoring of blood pressure and heart rate.

• It is used for EEG monitoring.

• Measuring blood glucose level.

Application

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Whole body plethysmography experiments are widely used in basic and preclinical research for non-invasive pulmonary measurements of conscious and unrestrained animals.

Plethysmography

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It is used to measure change in the lung volume.

It is used to determine the thoracic gas volume and air way resistance.

Used to measure the tidal volume.

Used to detect the asthma.

Application

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Microarray

Western blot

Flow cytometry

Biochemistry

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• A large number of DNA probes are present, each one of different sequence are immobilized at defined position on solid surface either made up of glass or nylon.

• For the preparation of high density arrays, oligo-nucleotide are synthesized in situ on the surface of glass or silicon.

Microarray Technique

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Working

Take tissue sample of any part of body organ

Extract the mRNA from cell and label with fluorescent dyes

Spread labeled mRNA on microarray plate contains 1000 of DNA

mRNA binds to DNA called hybridization. This hybridized plate ready to scan.

Scan the plate and analyze the data.

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Green color for the normal DNA.Red color for the tumor DNA.Yellow color equally expressed in tumor and normal DNA.

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This technique is used to examine the activity of various genes at different times.

This technique is used for determination of various types of cancer.

It also used for detection of heart disease, mental illness and infectious diseases.

Application

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Western blot is used in research to separate and identify proteins. In this technique a mixture of proteins is separated based on molecular weight, and thus by type, through gel electrophoresis.

These results are then transferred to a membrane producing a band for each protein. The membrane is then incubated with labels antibodies specific to the protein of interest.

Western Blotting

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For HIV detection

For detection of Lyme disease

For detection of Hepatitis B Virus

For detection of herpes

Application

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Flow cytometry is a technology that is used to analyze the physical and chemical characteristics of cells as it passes through at least one laser.

Cell components are fluorescently labeled and then excited by the laser to emit light at varying wavelengths.

Flow Cytometry

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Flow chamber

Laser

Light detector

Filters

Color detectors

Working

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Immunology: Identification of immune cells.

Neuroscience: For characterization of Alzheimer and Parkinson's disease.

Applications

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Micro ultrasonic imaging

Magnetic resonance imaging

Micro PET

IMAGING

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Confocal Microscopy

Micro ultra sound imaging

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Micro Ultra Sound Imaging

Ultra sound produces high resolution, rapid frame rate, low cost.

It is used for assessments of anatomic features blood flow in large vessels and measurement cardiac function.

Micro markers are used for visual imagingUltrasound in pre-clinical small animal

research by creating high-frequency transducers that offers superior high resolution (down to 30um). 

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Piezoelectric element produce caustic pulses above 20khz generate sound waves

Diverse tissue or organ reflects sound wave according to their density .

Gray scale produce images when reflected sound waves returns to the transducer.

Resulting image is viewable are produced instantly and captured a still photograph or movie

Working

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1. Non invasive, non radioactive detection of microtargetes

2. Less time required for investigational result

3. Easily translate the calculations

Advantages

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Developmental biology• In developmental biology it is useful for

assessing maternal, placental and fetal hemodynamic morphology and growth

Cancer• Tumor sizing and quantifying in two and

three dimensional images.• Quantification of tumor blood flow. Cardiovascular imaging• Quantification left ventricular functioning.

Applications

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Magnetic Resonance Imaging

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Magnetic Resonance Imaging

• Well known system for imaging of human body.

• Basically works on the principle of magnetic field.

• Use full to identify the damage parts of the body or tissue.

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WorkingBody is made up of the water which consists of H and O2. at

the center of hydrogen protons are present.

When body pass through the magnetic field the protons are line up in same direction magnet can pull them.

Short radio waves are send to the body knocking the proton out of alignment, radio waves turn off this proton realign and send

the radio signals picked by receivers.

The signals provides information about exact location of the proton and image is generate.May 3, 2023 29

• Morphological imaging

• Functional imaging

• Molecular imaging

Application

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Positron Emission Tomography (PET)

PET scan uses radiation to create 3D, Colour image of the functional organs.

They are used along with X-rays an MRI scan to develop images .

The PET scan looks functions of the organs

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WorkingA radio active medicine is used along with the natural

chemical(Glucose)

When it goes to the body area where glucose use for energy

PET scan detects the energy emitted by the positrons

As the radio tracer broken in body positrons made this image as 3D image

The image revels how parts of body function by way they break down the radiotracer

A PET image will display different levels of positrons according to brightness and colorMay 3, 2023

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Investigational studies

1. Epilepsy : It revel which part of brain is being affected or damaged.

2. Heart diseases : It helps to detect which part of heart is damaged or scared.

Application

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• It is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of adding space pin hole place at the confocal plane of the lens to eliminate out of focus light.

• It helps to reconstruction of three dimentional structure from obtained image.

Confocal Microscopy

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Working

Schematic diagram of confocal microscopeMay 3, 2023

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Catheters

Surgery

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Pressure volume catheter• Use to study hemodynamic analysis. • To study ventricular pressure and

volume in rat.• Osmotic pressure pump.

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Applications

Evaluating Pericarditis Using Left & Right Ventricular Pressure Waveforms

Pulmonary Artery Pressure (PAP) is one of the most commonly measured parameters during a cardiac catheterization case. Mean PAP, systolic PAP and diastolic PAP are often derived by visually marking the waveform output by a fluid-filled transducer.

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• Goldman M.D, Smith H.J, Ulmer W.T., (2005), “Whole body plethysmography” , Europe Respiratory Journal.

• Foster f., Hassack J., Adamson S., (2011). “Micro ultra sound for preclinical imaging”. Interface Focus vol. no. 1,576-601.

• Stout D. et al., (2013). “Guidance for methods description used in preclinical imaging papers”, Molecular imaging.

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References

• Catana C. et al.,(2008). Simultaneous in vivo positron emission tomography and magnetic resonance imaging, PNAS, volume 105, 3705-3710.

• Prasad v. et al.,(2007), Confocal microscopy of colloids, Journal of Physics.

• Satyanarayana U., Chakrapani U. Biochemistry, third edition,.589-593.

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• MRI Scan 2013, www.nhs.uk/conditions/MRIScan[Accessed 10/11/2014].

• PET Scan 2013, www.medicalnews-today.com/articles/154877/Hph[Accessed 1/11/2014].

• Flow cytometry,2014, www.unsolvedmystries.oregonstate.edu/f low_06[Accessed 8/11/2014].

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• Implantable telemetry www.itstelemetry.com/navigation/20bar/submenus/implantable_telemetry.php [Accessed 3/11/2014].

• Application of western blotting, www.western_blot.us/applications_ofwsternblotting/application_in_medical_diagnosis [Accessed 1/11/2014]

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THANK YOU