Predavanje 1 - Kompjuterizovanje merenje...
Transcript of Predavanje 1 - Kompjuterizovanje merenje...
Predavanje 1
Boris [aki}
Psychology Building Rm. 303, 1280 Main St. West Hamilton, Ontario, Canada L8S 4K1 e‐mail: [email protected] www.fhs.mcmaster.ca/psychiatryneuroscience/faculty/sakic www.brain‐body.ca/membership.html
Associate Professor
Department of Psychiatry and Behavioural Neurosciences McMaster University
My field of expertise: Behavioural Neuroimmunology PsychoNeuroImmunology 1. Lupus of the nervous system and role of brain-reactive
antibodies in etiology of aberrant behavior
2. Role of autoimmunity in etiology of Alzheimer’s disease
Structure of the 2-h class: Presentation 50 min Break 10 min Presentation / Discussions 50 min
Please pay particular attention to slides with pinky brain in the upper corner. They reflect a topic or knowledge I would like you to keep during your career.
Topic-relevant slides from my PowerPoint presentations will be uploaded in PDF format to the University server or your e-mail.
What is behavior?
From forest to trees
From a big picture to details
What is life?
Uniquely organized dead matter.
The matter cannot be destroyed, it merely changes forms.
Definition of Life
Compared to dead matter, life expands on our planet.
THREE MANDATES OF LIFE?
1. TO ADAPT IN CONSTANTLY CHANGING EXTERNAL ENVIRONMENT
2. TO SURVIVE COMPETITON FOR RESOURCES (VS. OTHER FORMS OF LIFE)
3. TO PROCREATE AND EXPAND THE SPECIES
BEHAVIOR
CONSTELLATION OF ACTIONS AND TRAITS
THAT PROMOTE THREE MANDATES OF LIFE
HOMEOSTASIS
• physical stimuli (light, sound, heat etc.) • psychological stress
Nervous System
Endocrine System
• Viruses/bacteria • Toxins • Oncogens • Autoantigens
Immune System
Behavior
Homeostatic Metasystem
cytokines
Hormones (e.g., steroids)
External stressors
Internal stressors
Behavior: adaptive (normal) or non-adaptive (abnormal)
Psychology Ethology
Psychiatry Experimental Psychiatry
ETHOLOGY – THE BIOLOGICAL STUDY OF BEHAVIOR
(HOW AND WHY)
PSYCHOLOGY – THE STUDY OF HUMAN BEHAVIOR
(PROXIMATE CAUSATION – HOW)
Why study behavior?
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MEASURING BEHAVIOR DURING NEUROINFLAMMATION
Each successive level of organization has properties that cannot be predicted even from complete knowing the lower level of organization. Detailed knowledge about behavioral phenomenon at molecular, cellular or physiological level is not helpful in fully understanding what is happening at the behavioral level.
Miseries of the Human Kind
1. Natural disasters (force majeure)
2. Physical and mental diseases (including addiction)
3. Aggressive behavior, violence, and wars
4. To understand how to live a happier and more productive life
1. To better understand the nature of animals, including homo sapiens
2. To understand the causes of mental illness and brain damage
3. To find new treatments for brain diseases
Why measure behavior?
PROBLEM CLASSIFICATION
PROXIMATE CAUSATION – HOW DO INTERNAL AND EXTERNAL CAUSAL FACTORS ELICIT AND CONTROL BEHAVIOR?
DEVELOPMENT – HOW DID THE BEHAVIOR DEVELOP DURING THE LIFETIME OF THE INDIVIDUAL?
FUNCTION – WHAT IS THE CURRENT USE OR SURVIVAL VALUE OF THE BEHAVIOR?
EVOLUTION - HOW DID THE BEHAVIOR ARISE DURING THE EVOLUTIONARY HISTORY OF THE SPECIES?
Three phases of life (according to ancient East Indian book of Kama Sutra)
3. Acquiring spirituality
1. Acquiring knowledge
2. Acquiring material things
Adaptive behavior - a principal instrument in achieving success in private and professional life
Schooling Employment Selection of a life partner Marriage Child upbringing Retirement
Principles of experimental science
1. Measurability
2. Controllability
3. Reproducibility
The assessment of behavior depends on:
1. Time of occurrence
2. Context
4. Sequence of behavioral acts
3. Intensity (latency, duration, frequency)
Behavior: Observed / descriptive - qualitative analysis
Systematically measured - quantitative analysis
Distribution of vertebrate animal species used for research, testing, and education
V Baumans Gene Therapy (2004) 11, S64–S66. doi:10.1038/sj.gt.3302371 (mice + rats 77%)
In 2010 – 26,000,000 laboratory animals used for research (mice + rats 95%)
The discovery of anaesthetics and Darwin's publication on the Origin of Species in 1859, defending the biological similarities between man and animal.
Claude Bernard published his book 'Introduction à l'étude de la médecine expérimentale' in 1865, introducing methodology as a tool for the design of physiological experiments.
The development of microbiology caused an increase in the use of animals, due to Koch's 'Postulates' where it is stated that pathogenicity of a microorganism can be proven after successfully infecting healthy, susceptible animals.
The development of pharmacology, toxicology and immunology.
Factors that led to increased use of animals in experimental research
Ethical standpoints
1. Absolutist (extremist) views: always justified, or never justified to use animals in biomedical research
1985 the Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (ETS 123) of the Council of Europe 1986 the Directive for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes of the EU, based on ETS 123, but more stringent.
http://www.apa.org/science/leadership/care/guidelines.aspx
2. Relativist view: certain uses are justified, but not all
Guidelines for Ethical Conduct in the Care and Use of Nonhuman Animals in Research
Benefits of animal research: medical advances that required animal research for their development Benefits to Humans Immunization against polio, diphtheria, mumps, measles, rubella, smallpox Antibiotics Anesthetics and other painkillers Blood transfusions Radiation and chemotherapy for cancer treatment Open-heart surgery for coronary bypass and correction of birth defects Insulin treatment for diabetes Asthma medication Medication for epileptic seizures Organ transplantation and drugs to prevent organ rejection Medications to treat depression, anxiety, and psychosis Microsurgery to reattach severed limbs Rehabilitation of stroke and brain-damaged accident victims Benefits to Animals Vaccination against rabies, distemper, tetanus, and other infections Corrective surgery for hip dysplasia in dogs Orthopedic surgery and rehabilitation for horses Treatment for leukemia and other cancers in pets Improved nutrition for pets Source: Foundation for Biomedical Research, 1988
Guiding principles of three R's: Replacement - substitution of living animals by in vitro techniques (e.g., cells, tissues), computerized models, etc. Reduction - decrease of the number of animals using power analysis, reduce experimental procedures and number of housed animals for improved well being Refinement - environmental enrichment, adequate anaesthesia, analgesia and care, by guaranteeing skills of the researcher /animal staff, by improving experimental procedures and by determining a humane end point, where the animal can be euthanized to prevent unnecessary suffering (CCAC, AUP)
Russell & Burch in 1959: “The Principles of Humane Experimental Technique”
1. Better controlled 2. More consistently measured 3. Shorter than clinical studies 4. Can test cause-effect relationships
Advantages of animal studies:
Development of animal use in the 20th century
V Baumans Gene Therapy (2004) 11, S64–S66. doi:10.1038/sj.gt.3302371
Drop due to: - public awareness - strict legislation on animal use - the development of animal ethics committees
Increase due to development of genetically modified animals
75–100 million vertebrates per year are used in research and testing
Distribution of the purposes for animal use
V Baumans Gene Therapy (2004) 11, S64–S66. doi:10.1038/sj.gt.3302371
1. How worthwhile is a proposed piece of research in terms of its likely contribution to scientific understanding?
2. How likely is the research to bring benefits for humans, animals, economy or environment?
3. How much suffering to animals is likely to result from the research?
Types of judgment to be made before doing an experiment
The Bateson decision cube
The Canadian Council on Animal Care: http://www.ccac.ca/en_ McMaster's Animal Research Ethics Board (AREB) http://fhs.mcmaster.ca/healthresearch/areb_introduction.html
Institutional Animal Care and Use Committee (IACUC)
The subjective experiences of an animal may be totally different from humans, reflecting its different way of life and the different ways in which its body works.
Human interpretation of what is observed in an animal should not be based only on what is known from humans, but also on a good knowledge of its natural history and behavior.
= /
Animal behavior = human behavior ?
Using human emotions and intentions as explanations for animals’ actions can impede further attempts to understand the behaviour.
Absence of evidence is not evidence for absence.
Anthropomorphism Anthropomorphism or personification is any attribution of human characteristics to animals, objects, and natural phenomena.
Lecture 1
Kompjuterizovano merenje pona{anja
Human vs. computerized scoring of behavior
Isolated tests vs. home-cage behavior
Category Factors Ontogeny intrauterine position, litter size, maternal behavior Housing transport to animal facility, size of housing cage, environmental enrichment, social vs. individual housing, light/dark schedule, vibrations / ultrasonic sounds, temperature, humidity, diet Test situation size, shape, and cleanness of test apparatus, protocol, time of testing, light intensity
Factors affecting reliability and reproducibility of behavioural data
Human handling during cage cleaning and transportation to a testing room, observer’s presence, scoring experience, fatigue, and awareness of group origin
Sampling (which subjects and behavioral categories are observed)
Ad libitum Focal Scan Behavior
Ad libitum = no systematic constraints are placed on what is recorded and when. Observer simply notes down whatever is visible and seems relevant at the time. Can be useful during preliminary observations.
Focal = observing one individual or a social unit (e.g., dyad, litter) and recording all instances of its behavior.
Scan = a whole group of subjects is rapidly scanned at regular intervals and the behavior of each individual at that instant is recorded. Often restricts the observer to recording only one or a few simple categories of behavior.
Behavior = watching the whole group and recording each occurrence of a particular type of behavior, together with details of which individuals are involved.
Recording (for how long the behavior is observed)
Continuous Time sampling
Instantaneous One-zero
Continuous = provides an exact and faithful record of behavioral patterns, measuring true frequencies, times, and durations.
Time sampling = sampling of behavior in intervals. Less information is preserved and an exact record of the behavior is not necessarily obtained.
Sample points
Sample intervals
Time
Start
occurrences / sample points
How good / right are your dependent measures? Reliability (“good”) - the extent to which the measurement is repeatable and consistent: that is, free from random errors Precision – how free is the measurements from random errors? Sensitivity – are small changes in the true value reflected by changes in the measured value? Resolution – what is the smallest change in the true value that can be detected? Consistency – do the repeated measurements of the same behavioural act produce the same results?
Validity (“right”) - the extent to which a measurement actually measures what the investigator wishes to measure and provides information that is relevant to the questions being asked. Accuracy – how free is the measurements from systematic errors? Specificity - to what extent does the measure describe what it is supposed to describe, and nothing else? 1. 3.
2. 4.
Human vs. computerized scoring of behavior
Company Country Website AccuScan Instruments, Inc. (1) U.S.A http://www.accuscan-usa.com
BioSignal Group Corp. (2) U.S.A http://www.biosignalgroup.com
Biobserve (1) Germany http://www.biobserve.com/experiments/fst-tst.html
Campden Instruments Ltd (1) UK http://www.campdeninstruments.com
Cleversys, Inc (1) U.S.A http://www.cleversysinc.com
Columbus Instruments (1) U.S.A http://www.colinst.com
Coulborn Inc (1) U.S.A http://www.coulbourn.com
HVS Image (2) UK http://www.hvsimage.com/index.htm
IITC Inc (3) U.S.A http://www.iitcinc.com/whatsnew.html
Kinder Scientific (3) U.S.A http://www.kinderscientific.com
Lafayette Instruments Co. Inc (1) U.S.A http://www.lafayetteneuroscience.com
MED Associates Inc. (1) U.S.A http://www.med-associates.com
NewBehaviour (1) Switzerland http://www.newbehavior.com
Noldus Information Technology (1) The Netherlands http://www.noldus.com
Panlab, S. L. (1) Spain http://www. panlab.com
San Diego Instruments Inc (1) U.S.A http://www.sandiegoinstruments.com
Stoelting Inc (1) U.S.A https://www.stoeltingco.com
TSE Tech Sci. Equipment GMBH (1) Germany http://www.tse-systems.com
(1): Limited video-tracking / event recording in home-cage environment; (2): Video-tracking only; (3): Limited hardware-based tracking out of home-cage environment.
Major software / hardware manufacturers of behavioral equipment
Reliable continuous measurements became feasible due to: 1. Increased speed of computer processors 2. Development of digital video formats 3. Increased capacity of storage media 4. Development of real-time video tracking
Computer > Human observer
From time sampling to continuous recording
1. Counting of behavioral responses (e.g., after drinking, eating, wheel running)
2. Position tracking of experimental animals (distribution of locomotion)
3. Classification of behavioral acts (e.g., grooming, sleeping, yawning, etc.) Human-based vs. Software-based applications)
Human observer > Computer
Computer > Human observer
Domains of computerized data acquisition
The list of behaviors in Ethogram Giant Panda
Manual counting of behavioral acts
Computerized counting of behavioral outputs
MedPC IV from Med Associates Inc.
Position tracking
Manual
Video-tracking (center of gravity, 3-point detection)
Infra-red technology
Infra-red technology
Video-tracking
Video-tracking with EthoVision XT 8
EthoVision XT – Zone Setting / Calibration
EthoVision XT – Data Acquisition
EthoVision XT – Data Acquisition in Multiple Arenas
Software-based
Automated classification of behavioral acts
Human-based
Observer XT
CleverSys
Behavioral Instruments (BI)
Analytical technique that identifies the behavior of organisms based on positional and vibratory information
Behavioral Spectroscopy
Automated Seizure Detection in Mice
Automated Forced Swim Assay
Multiple tests vs. monitoring of home-cage behavior
1. Assume a single behavioral domain or category 2. Restricted to a novel environment 3. Short lasting
Disadvantages of “standard” behavioural tests:
Relevant dynamic processes, such as circadian, monthly, and annual rhythms, are not included.
Behavior is a process, not an event.
Important information on baseline conditions and interactions between motivational systems cannot be obtained.
Neuron, Vol. 41, 825–837, March 4, 2004
1. Allows continuous measurements of both baseline behaviour and experimental manipulations.
2. By introducing stimuli to elicit specific behavioural responses, multiple behavioural domains and their interactions can be addressed in a single test set-up.
3. Minimalized stress induced by handling and transportation (confounding factors).
4. Use an standardized testing environment, thereby reducing between-laboratory variance.
5. Fully automated home cage system is time and cost saving, minimizes the need for human intervention and biased analysis.
Advantages of home cage testing
1. Enables video recording independent of light conditions in the laboratory. 2. Enhances the standardization of behavioral experiments. 3. Extendable with specially designed home cage accessories and integrated visual and acoustic stimuli. 4. PhenoTyper offers the test set-up for activity and open field studies, interactive home cage testing, and circadian studies.
Phenotyper by Noldus
Phenotyper
Phenomaster by TSE
social interaction cage (placed when required)
Novel object (placed when required)
running wheel (CT-ENV-043A)
PC with EthoVision XT5,Observer XT, MedPC software packages
stimulus light (ENV-221M)
climbing mesh (CT)
removable tray
water (photobeam lickometer ENV-351W)
Food dispenser -20 mg pellets (ENV-203-20)
digital interface package (DG-716P2)
sucrose solution (photobeam lickometer ENV-351W)
Picolo Diligent BNC card (live tracking and/or MPG4 video recording)
Infra-red Panasonic WV-CP484 camcorder
Integrated Behavioral Station (InBeSt) designed by B. Sakic
Shelter (see through) (IR-010 Infrared Plastic)
Molex cable
BNC cable
INBEST = EthoVision + Med PC IV
Variables: MedPC IV Total Number of Licks on Lickometer 1 (water intake) Latency to First Lick on Lickometer 1 Total Number of Licks on Lickometer 2 (sucrose intake) and volume ingested (manually recorded) Latency to First Lick on Lickometer 2 Total Number of Food Pellets Taken Latency to First Food Pellet Taken Total Number of Counts on Running Wheel Total Number of Runs on Running Wheel Total Time Spent on Running Wheel Latency to Running Wheel Total Number of Climbs on Climbing Mesh Total Time Spent on Climbing Mesh Latency to Climbing Mesh from EthoVision XT 8 Number of entries into the shelter Time spent in the shelter Distance traversed, moving time, locomotor speed Number of visits to the running wheel Latency to approach a novel object Exploration of a novel object (frequency, duration) Sleeping episode (duration, frequency) Grooming (duration, frequency) Seizures (duration, frequency)
InBeSt
Variables used in behavioral assessments
MedPC + EthoVision
Summary Benefits of computerized analysis of behavior
Computerized methodology led to: 1. increased reliability of behavioral measures 2. improved standardization of behavioral protocols 3. minimized confounding factors associated with scoring by human
observer 4. high-throughput phenotyping Automated home cage monitoring allowed: 1. minimized exposure to stressors associated with multiple tests 2. measurement of behavior is conducted in ethologically relevant
context