Food Protection Trends - November/December 2017 - 435

transcripts was published recently (19). The establishment
of biomarker signatures of hepatotoxicity in vitro is the
goal of a current BMBF (German Federal Ministry for
Education and Research)-funded research project that
is being conducted with the involvement of the German
Federal Institute for Risk Assessment (http://www.bfr.
bund.de/de/modellierung_des_toxoms_ kultivierter_
menschlicher_hepatozyten__livsys_-193087.html).
On the basis of possibilities that already exist for
identifying certain effects via transcriptional signatures,
it is therefore conceivable in principle that test systems
could be developed that either enable the measurement
of a single biomarker set as an indicator of a certain effect,
or that are suitable at the same time for determining several
biomarker sets for the parallel identification of more
than one molecular mechanism of action. From a methodological point of view, depending on the numbers of
transcripts to be analyzed, standard RT-PCR methods,
so-called low-density arrays that combine a number of
target transcripts, or whole-transcriptome microarrays,
might come into use. Provided that appropriate specific
mRNA signatures independent of one another could
be found for various mechanisms of action, a system of
this kind would in principle be at least comparable to a
reporter gene battery, but could be conducted in only a
single experiment in a solitary biological system (Fig. 1C).
An analysis of this kind is without doubt comparatively
complex, as it involves sample processing and extraction,
cell incubation, RNA isolation, PCR and data evaluation.
The possibility of covering in parallel and at a reasonable
cost a large number of endpoints that would otherwise have
to be determined separately could nevertheless make the
execution of such a method appear feasible from a practical
point of view.
Issues and challenges to be solved for routine
application of transcript-based methods
There can be no doubt that a great deal of research still
has to be done before transcript-based approaches can be
put to use in routine operations in food and feed control.
The challenges to be faced include the following:
* As the basis for development and application of effectbased analytical test systems, knowledge of molecular
toxicity mechanisms has to be further widened. This
includes the mechanistic analysis of the interactions of
various mechanisms of action that could be expected in
the event of co-exposure of a cell to mixtures of two or
more chemicals with different effects.
* In order to establish multi-endpoint systems, i.e.,
biological test systems that permit simultaneous
detection of several effects, the functionality of the
identified molecular toxicity mechanisms has to be
verified experimentally in each test system. Many cells
are known to alter their properties as a consequence of
transition into in vitro culture.

* The identification of specific biomarker signatures
for individual molecular mechanisms of toxicity
is a complex task requiring the interdisciplinary
cooperation of toxicologists and bioinformaticians.
* The establishment of dose-response curves for signatures of several transcripts that may under certain
circumstances be of different strengths or be regulated
with different sensitivities also requires the inclusion
of bioinformatic expertise.
* The majority of the laboratories that routinely monitor
foods and feeds do not have the necessary prerequisites, with regard to technical equipment and
specialized knowledge, to use biological test systems,
including subsequent analysis at the transcript level.
Efforts are required here to create the necessary
infrastructure and expertise.
* Efforts are also required in the field of sample processing. Extraction and other methods are needed to
ensure that the resultant samples do not have any toxic
effects on the biological systems used, due to solvents,
for example, and also to ensure that if a large number
of effects that can be caused by structurally different
chemicals are to be examined, all of the different types
of chemicals are in the extract and do not get lost in
the course of processing.
* In general, comprehensive validation has to be conducted
before a new method in food and feed monitoring can be
used, and one would also have to pay due consideration
to the variability of biological systems.
NON-TARGETED EFFECT-BASED ANALYTICS ON
A TRANSCRIPTOME BASIS
The approaches outlined previously aim at the detection
of specific effects and signal modulations, and at the
possible widening of methods of this kind through the
parallel determination of several different endpoints or
mechanisms of action, each of which should be as specific as
possible. All of these would lie within the range of targeted
analytics. If it is assumed that biological effects are reflected
sensitively in the transcript pattern of a cell and that a
large number of mechanistically different effects exist, this
means ultimately that in principle, all mechanisms of action
could be recorded in parallel by recording the expression
change of all transcripts of a cell, at least to the extent
that the selected biological model system was capable of
reproducing them. Transferred to the concept of an "effectorientated" as opposed to a mechanism-based approach,
in the sense of non-targeted analytics without regard to
specific signaling pathways influenced by a substance, a
"degree of biological deregulation" could be determined
as a measure of the disturbance of cellular equilibrium by
exogenous substances, i.e., a parameter determined by the
number of deregulated transcripts as well as the amplitude
of their deregulation. Compared with the relatively insensitive effect-based endpoint of cytotoxicity in vitro, a paraNovember/December Food Protection Trends

435


http://www.bfr.bund.de/de/modellierung_des_toxoms_ kultivierter_ menschlicher_hepatozyten__livsys_-193087.html http://www.bfr.bund.de/de/modellierung_des_toxoms_ kultivierter_ menschlicher_hepatozyten__livsys_-193087.html http://www.bfr.bund.de/de/modellierung_des_toxoms_ kultivierter_ menschlicher_hepatozyten__livsys_-193087.html

Table of Contents for the Digital Edition of Food Protection Trends - November/December 2017

The Cleanliness of Resusable Water Bottles: How Contamination Levels are Affected by Bottle Usage and Cleaning Behaviors of Bottle Owners
Impact of Carcass Anatomical Location on the Microbiological Profile of Beef Trimmings
Thermal Processing Parameters to Ensure a 5-log Reduction of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in Acidified Tomato-based Foods
Experimental Evaluation of Performance of Sampling Techniques for Microbiological Quantification on Carcass Services
Effect-based Analytics for Toxicological Screening - Concepts for Future Developments
Beyond the Bio - John Luchansky
PDF Highlight - Food Chemical Hazards and Food Allergy PDG
IAFP 2017 In Review
Industry Products
Coming Events
Food Protection Trends - November/December 2017 - Cover1
Food Protection Trends - November/December 2017 - Cover2
Food Protection Trends - November/December 2017 - 385
Food Protection Trends - November/December 2017 - 386
Food Protection Trends - November/December 2017 - 387
Food Protection Trends - November/December 2017 - 388
Food Protection Trends - November/December 2017 - 389
Food Protection Trends - November/December 2017 - 390
Food Protection Trends - November/December 2017 - 391
Food Protection Trends - November/December 2017 - The Cleanliness of Resusable Water Bottles: How Contamination Levels are Affected by Bottle Usage and Cleaning Behaviors of Bottle Owners
Food Protection Trends - November/December 2017 - 393
Food Protection Trends - November/December 2017 - 394
Food Protection Trends - November/December 2017 - 395
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Food Protection Trends - November/December 2017 - 399
Food Protection Trends - November/December 2017 - 400
Food Protection Trends - November/December 2017 - 401
Food Protection Trends - November/December 2017 - 402
Food Protection Trends - November/December 2017 - Impact of Carcass Anatomical Location on the Microbiological Profile of Beef Trimmings
Food Protection Trends - November/December 2017 - 404
Food Protection Trends - November/December 2017 - 405
Food Protection Trends - November/December 2017 - 406
Food Protection Trends - November/December 2017 - 407
Food Protection Trends - November/December 2017 - 408
Food Protection Trends - November/December 2017 - Thermal Processing Parameters to Ensure a 5-log Reduction of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in Acidified Tomato-based Foods
Food Protection Trends - November/December 2017 - 410
Food Protection Trends - November/December 2017 - 411
Food Protection Trends - November/December 2017 - 412
Food Protection Trends - November/December 2017 - 413
Food Protection Trends - November/December 2017 - 414
Food Protection Trends - November/December 2017 - 415
Food Protection Trends - November/December 2017 - 416
Food Protection Trends - November/December 2017 - 417
Food Protection Trends - November/December 2017 - 418
Food Protection Trends - November/December 2017 - Experimental Evaluation of Performance of Sampling Techniques for Microbiological Quantification on Carcass Services
Food Protection Trends - November/December 2017 - 420
Food Protection Trends - November/December 2017 - 421
Food Protection Trends - November/December 2017 - 422
Food Protection Trends - November/December 2017 - 423
Food Protection Trends - November/December 2017 - 424
Food Protection Trends - November/December 2017 - 425
Food Protection Trends - November/December 2017 - 426
Food Protection Trends - November/December 2017 - 427
Food Protection Trends - November/December 2017 - 428
Food Protection Trends - November/December 2017 - 429
Food Protection Trends - November/December 2017 - Effect-based Analytics for Toxicological Screening - Concepts for Future Developments
Food Protection Trends - November/December 2017 - 431
Food Protection Trends - November/December 2017 - 432
Food Protection Trends - November/December 2017 - 433
Food Protection Trends - November/December 2017 - 434
Food Protection Trends - November/December 2017 - 435
Food Protection Trends - November/December 2017 - 436
Food Protection Trends - November/December 2017 - 437
Food Protection Trends - November/December 2017 - Beyond the Bio - John Luchansky
Food Protection Trends - November/December 2017 - 439
Food Protection Trends - November/December 2017 - 440
Food Protection Trends - November/December 2017 - PDF Highlight - Food Chemical Hazards and Food Allergy PDG
Food Protection Trends - November/December 2017 - 442
Food Protection Trends - November/December 2017 - 443
Food Protection Trends - November/December 2017 - 444
Food Protection Trends - November/December 2017 - IAFP 2017 In Review
Food Protection Trends - November/December 2017 - 446
Food Protection Trends - November/December 2017 - 447
Food Protection Trends - November/December 2017 - 448
Food Protection Trends - November/December 2017 - 449
Food Protection Trends - November/December 2017 - 450
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Food Protection Trends - November/December 2017 - 530
Food Protection Trends - November/December 2017 - 531
Food Protection Trends - November/December 2017 - 532
Food Protection Trends - November/December 2017 - 533
Food Protection Trends - November/December 2017 - Industry Products
Food Protection Trends - November/December 2017 - 535
Food Protection Trends - November/December 2017 - 536
Food Protection Trends - November/December 2017 - 537
Food Protection Trends - November/December 2017 - 538
Food Protection Trends - November/December 2017 - 539
Food Protection Trends - November/December 2017 - 540
Food Protection Trends - November/December 2017 - 541
Food Protection Trends - November/December 2017 - 542
Food Protection Trends - November/December 2017 - 543
Food Protection Trends - November/December 2017 - Coming Events
Food Protection Trends - November/December 2017 - Cover3
Food Protection Trends - November/December 2017 - Cover4
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