Test format: Open book exam
Date and time: Friday Sept 30, 8:45-9:45 h
Pleases email your answers to aswin.mangerich@uni-konstanz.deuntil
10 am.
If possible, please write your answers in
red color.
Overview of toxicity testing and the 3 Rs concept:
requirements, goals & status quo (Mangerich
/Leist)
Question 1 [3
points]
Please name three advantages and
limitations/disadvantages, respectively, of the use of animals in toxicity
testing.
Question 2 [4
points]
Below,
eight non-standard (3R) approaches to assess hazard are given (A-H). Decide for
each approach whether it is based on the use of animals or not. Animal-based
means that animals will or may be used. Non-animal means that live animals are
not used. Mark your choice of the correct answer with a cross in either the 3rd
or 4th column. Each correct answer gives 0.5 points. Wrong answers
give 0.5 negative points. The maximum number of points is four, the minimum is
zero.
|
Approach |
Name |
animaluse |
noanimaluse |
|
A |
Reduction |
X |
|
|
B |
Refinement |
X |
|
|
C |
Replacement |
|
x |
|
D |
In vitro NAM |
|
x |
|
E |
In silico NAM |
|
x |
|
F |
QSAR |
|
x |
|
G |
Readacross |
|
x |
|
H |
IATA |
x |
|
Introduction into test development for hazard and
toxicokinetic predictions (Leist)
Question3 [5 points]
Below,
five statements are made which are to be judged as TRUE or FALSE. Mark your
choice of the correct answer with a cross in either the 2nd or 3rdcolumn.
Each correct answer gives 1 point. Wrong answers give 1 negative point. The
maximum number of points is five, the minimum is zero.
|
Statement |
True |
False |
|
The PoD
for a NAM (e.g. hepatotoxicity of chemical A) is given in mg/kg |
|
x |
|
The clearance of compound A in hepatocytes is determined
usually by measuring the half-life of compound A, when incubated with
hepatocytes |
x |
|
|
PBK models require parameters for the transition of chemicals between
(body) compartments |
x |
|
|
A NAM is always
setup to predict chemical hazard and it can therefore be used for hazard
determination of any chemical |
|
x |
|
Each cell-based NAM
has a defined test endpoint (e.g. ATP content) |
x |
|
The concept of adverse outcome pathways (AOPs)
(Schildknecht)
Question 4 [2
points]
Provide a brief definition of the term“Adverse
Outcome Pathway (AOP)”.
Question 5 [2
points]
What are 2 key requirements that qualify a biological
state as “Key Event (KE)”
Validation concepts and test readiness criteria
(Waldmann)
Question 6 [5
points]
Below, five statements are made which are to be judged
as TRUE (T) or FALSE (F). Mark your choice of the correct answer with a cross
in either the 2nd or 3rd column. Each correct answer gives 1 point. Wrong
answers give 1 negative point. The maximum number of points is five, the
minimum is zero.
|
True |
False |
|
|
The solvent control is
the only negative control that is needed for validation of a test method |
|
x |
|
The
endpoint-specific controls are used to gain confidence that pathways
considered to be relevant for the endpoint are indeed affected. |
x |
|
|
The prediction model can be also a more complex decision tree to
combine two endpoints or even several test methods |
x |
|
|
Acceptance criteria must be set before the first test runs for test
method validation. |
x |
|
|
To define acceptance criteria historical data must be collected in order
to learn about baseline variations and test performance. |
x |
|
Good in vitro method practices guidance; a tool to
increase trust in data for (regulatory) decision making (Langezaal)
Question 7 [1
point]
Which action described below is recommended by GIVIMP.
Please check the correct answer.
a)
In-house validation of
the in vitro method, after the
generation of data and publication of results.
b)
Inclusion of positive
and negative controls, to demonstrate that the test system (e.g. cells, tissue
or cellular fraction) responds as expected. -
c)
Setting cut-off limits
for the test item results below the limit of detection.
d)
Use of fetal bovine
serum and antibiotics to enhance cell attachment and suppress
microbiological infections, respectively.
Question 8 [1
point]
Which of the following elements is NOT part of
an in vitro method
a)
the test system (e.g.
a cell or tissue)
b)
positive and negative
control items
c)
data analysis and
data interpretation model
d)
the test item
New approach methodologies (NAMs) in hepatotoxicity
testing (Hengstler)
Question 9 [4
points]
Please assign the cell types of the
liver below to a typical function:
Cell type of the liver
(a) Hepatocyte 2
(b) Kupffer cell 3
(c) Sinusoidal endothelial cell 1
(d)
Stellate
cell 4
Function
1. First contact with immune cells of the
blood
2.
Metabolism
3.
Phagocytosis
of bacterial fragments
4.
Secretion
of extracellular matrix
History and state of the art of stem cell-based
toxicity testing (Seeger)
Question 10 [2
points]
Which aspects make the fetus
extremely sensitive to developmental toxicants?
Mark
your choice of the correct answer with a cross in either the 2nd or
3rd column. Each correct answer gives 0.5 points. Wrong answers give
0.5 negative points. The maximum number of points is 2 points, the minimum is
zero points.
|
Statement |
Yes |
No |
|
Most rapid cell
growth/organ differentiation |
x |
|
|
Organs are fully differentiated |
|
x |
|
High degree of epigenetic
plasticity |
x |
|
|
Tight blood brain barrier |
|
x |
Question 11 [ 1 point]
Which would be the perfect
culture method for stem cells concerning Good Cell Culture Practice (one answer
is correct)? c
a) feeder-dependent, xenogenic, chemically undefined
b) feeder-independent, xenogenic, chemically undefined
c) feeder-independent,
xenofree, chemically defined
d) feeder-dependent,
xenofree, chemically defined
Read-across for the prediction of toxiceffects
of chemicals (vomBrocke)
Question 12 [4
points]
Name four aspects that belong to every read-across
approach?
Computational toxicology methods for prediction of
preclinical safety endpoint (Anger)
Question 13 [2
points]
What does the fundamental “Similarity principle” in
Computational Toxicology mean?
Question 14 [1
point]
Bacterial mutagenicity, an endpoint for which many
reliable in silico prediction models exist, is mainly driven by...
a) by physico-chemical properties
b) through primary (on-)target protein activity
c) by reactive chemical substructures
d) through secondary (off-)target protein activity
Use of zebrafish as a model organism in the context of
3Rs (Scholz)
Question 15 [1
point]
Why are zebrafish embryos considered as an alternative
model and 3R-compliant?
Question 16 [1 point]
For which mode of action the zebrafish has reduced
sensitivity in mortality if compared to adult fish?
Non-animal approaches in genetic toxicology and
carcinogenicity testing(Martus/Mangerich)
Question 17 [2 points]
Which 2 key strategies are followed in order to
further reduce animal numbers in genotoxicity testing?
Question 18 [2 points]
What are the advantages of using transgenic animals as
opposed to wildtype animals? Please check the correct answer(s)
A1: shorter treatment duration
A2: higher sensitivity
A3: higher relevance of tumor findings
A4: less animals
Microphysiological
systems (MPS) and organ on a chip (OoaC)Technologies
(Loskill)
Question 19 [2 points]
Which of these aspects are basic principles of
Organ-on-Chip models? Please check the correct answers.
a. Physiological
microenvironment
b. Culture
of microorganisms
c. Integration
of electrodes
d. Vacuum
channels
e. Microfluidic perfusion
Question 20 [3 points]
What are advantages of organoid models? Please check
the correct answers.
a. Human
genetic background
b. Cell
polarization
c. Physiological
biochemical and biophysical cues
d. Static
culture
e. Cell
heterogeneity
New approach methodologies (NAMs) in
(developmental)neurotoxicity (DNT) testing (Fritsche)
Question 21 [3 points]
Name at least three cellular readouts for DNT.
New developments in reduction & refinement of
animal experiments (Würbel)
Question 22 [3 points]
What do the 3Vs stand for and what is their role in a
proportionality test of animal research?
European regulatory landscape of 3 Rs:Why
Non-Animal Methods are controversial in EU policies? (Busquet)
Question 23 [4 points]
Name (a) the institutions involved in EU policy making
and (b) at least 2 directorates-general (DG) involved in EU policies on the use
of laboratory animals.
Relevance of new approach methodologies (NAMs) for the
chemical industry - today and tomorrow (Birk)
Question 24[3 points]
What are critical requirements on NAMs during the
development process – please list at least three requirements?
Development of OECD-approved test guidelines for new
approach methods for skin sensitization (Kolle)
Question 25 [3 points]
Borderline range around prediction cut-offs. Check all
answers that are correct.
a) Any toxicological test result can clearly be
assigned to the effect or no effect side.
b) A borderline result can come from two major sources
including experimental variability or inherent properties of the item tested.
c) Using binary “hazardous/non-hazardous” information may
lead to ambiguous classifications of substances
d) Quantifying the area where such ambiguous results
are likely to occur, i.e. the borderline range, can be used to document a
testing method’s limited precision
3Rs in the context of food safety (Kass)
Question 26 [2 points]
What are the main perceived obstacles to the use of
NAMs in regulatory risk assessment for food safety?
The concept of adverse outcome pathways (AOPs) (Schildknecht)
Question 4 [4 points]
Please give a brief definition of the AOP terms:
“Key Event”
“Key Event Relationship”
Good in vitro method practice guidance, test readiness
criteria and validation concepts (Langezaal)
Question 5 [1 point]
Which of the following is true:
a) The use of test readiness criteria is obligatory to
confirm that a method is ready for validation
b) The use of test readiness criteria is helpful to
understand if method development is complete
c) Test readiness criteria are an official tool of
GD34, used by peer-reviewers, to confirm validation is complete
d) None of the above
Question 6 [1 point]
What needs to be ensured, before a NAM will be used
for regulatory decisions?
a) All data
generated during validation meet the acceptance criteria.
b) The
method developer confirms that the method is fit for purpose, biologically
relevant and technically characterized.
c) Chemicals
that trigger the mode of action cannot be identified.
d) Biological
relevance, robustness, accuracy, reproducibility, and fitness for a specific
purpose.
Systems Toxicology (Hartung)
Question 7 [2 point]
Which omics technologies are mainly used in systems
toxicology?
History and state of the art of stem cell-based
toxicity testing (Seeger)
Question 8 [3 points]
Name three types of stem
cells. Give information in bullet points about the source and potency of each type.
Use of zebrafish as a model organism in the context of
3Rs (Scholz)
Question 10 [1 point]
Which mode of action can be detected by assessment of
embryonic movements?
Computational toxicology methods for prediction of
preclinical safety endpoint (Anger)
Question 13 [2 points]
What's the goal of applying Computational Toxicology
methods in the early stages of drug discovery?
a) Risk assessment
b) High throughput
c) Prediction of toxic potential ("hazard")
d) Low throughput
New approach methodologies (NAMs) in (developmental)
neurotoxicity (DNT) testing
(Fritsche)
Question 17 [4 points]
The DNT in vitro battery...
1. Consists of in vitro assays that cover basic
neurodevelopmental processes.
2. Is a collaborative approach involving regulators
and the OECD.
3. Only contains human cell systems.
4. Has been scientifically validated.
5. Is a continuing effort that takes novel
developments into account.
Please mark the correct answers.
3Rs from the Pharma perspective (Moretti)
Question 19 [2 points]
Can you name two reasons behind using alternative
methodologies in investigative toxicology?
European regulatory landscape of 3 Rs: Why Non-Animal
Methods are controversial in EU policies? (Busquet)
Question 21 [2 points]
List min two reasons why NAMs are controversial?
Relevance of new approach methodologies (NAMs) for the
chemical industry - today and tomorrow (Kolle)
Question 22 [3 points]
Industry use NAMs for different purpose. Please name
three examples.
3Rs from the BfR perspective
(Schönfelder)
Question 23 [1 point]
PARERE has a number of roles to fulfil: (Please mark
the correct answer)
1. To
provide upstream input on potential regulatory relevance and suitability of
proposed test methods and testing strategies.
2. Facilitate
information flow between OECD and regulators regarding the development and
validation of methods and to identify areas that need specific attention.
3. Identification
of specifically industry partners to participate in specific EURL ECVAM project
groups (e.g. ITS design, validation management groups, expert workshops etc.).
4. Provide
comments on draft EURL ECVAM recommendations following OECD peer review of
validation studies.
5. Support
and promote the role of OECD laboratories to facilitate their participation as
a testing and/or lead laboratory in EURL ECVAM led validation studies.
Read-across for the prediction of toxic effects of
chemicals (vom Brocke)
Question 24 [3 points]
What are potential advantages of read-across over
other test and non-test methods? Mention at least three:
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Last Update: 12. Oktober 2023