Supporting Publications 2012:EN-232
SCIENTIFIC REPORT submitted to EFSA
Investigation of the state of the science on combined actions of chemicals in
food through dissimilar modes of action and proposal for science-based
approach for performing related cumulative risk assessment1
Prepared by:
Andreas Kortenkamp (ULSOP), Richard Evans (ULSOP), Michael Faust
(F+B), Fritz Kalberlah (FoBiG), Martin Scholze (ULSOP), Ulrike
Schuhmacher-Wolz (FoBiG)
(ULSOP: School of Pharmacy, University of London; F+B: Faust and Backhaus
Environmental Consulting GbR; FoBiG: Forschungs- und Beratungsinstitut Gefahrstoffe
GmbH)
1
Question No EFSA-Q-2010-00866. Accepted for Publication on 23 January 2012.
Any enquiries related to this output should be addressed to pesticides.pprprocurement@efsa.europa.eu
Suggested citation: Kortenkamp A, Evans R, Faust M, Kalberlah F, Scholze M and Schuhmacher-Wolz U. Investigation of
the state of the science on combined actions of chemicals in food through dissimilar modes of action and proposal for
science-based approach for performing related cumulative risk assessment. Supporting Publications 2012:EN-232. [233
pp.]. Available online: www.efsa.europa.eu/publications
© European Food Safety Authority, 2012
Combined actions of chemicals in food through dissimilar modes of action
Abstract
The purpose of this project was to summarise the state of the science on combined actions of
chemicals in food through dissimilar modes of action and to propose a science basedapproach for performing the related cumulative risk assessment (CRA).
A systematic literature search was carried out (Task 1) to identify relevant experimental
studies of mixtures, and this served as the basis for a summary of the state of the science
(Task 2). Of central importance was the confirmation that there is no current example of a
situation in which the concept of independent action (IA) provides an accurate prediction that
is also more conservative than dose addition (DA), supporting the use of DA as a
conservative default in CRA. The quantitative difference between predictions based on DA
or IA were analysed in detail, and this analysis suggested that the differences that might be
expected in practice are small.
Currently used approaches to grouping (Task 3) and to CRA (Task 4) were reviewed to
identify their critical features. An approach is proposed (Task 5) that unifies the assessment
of similarly and dissimilarly acting chemicals based on pragmatic use of assessment
approaches derived from the concept of DA. The approach incorporates a tiered framework.
At lower tiers, the grouping of chemicals is driven by their co-occurrence in the exposure
scenarios under investigation. At higher tiers, chemicals that evoke a common adverse
outcome should be grouped together. The proposal is illustrated with three case studies.
Conclusions and recommendations
It is feasible and justified to utilise CRA methods and tiered framework analyses derived
from DA also for combinations of dissimilarly acting chemicals. There can be one unified
approach for dealing with mixtures in regulatory practice, irrespective of (often presumed)
modes of action.
Key words:
Combined actions of chemicals, cumulative risk assessment, dissimilar modes of action, dose
addition, science-based approach, independent action
Supporting publications 2012:EN-232
2
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Summary
The aims of this project were to 1) summarise the state of the science on combined actions of
chemicals in food through dissimilar modes of action, and 2) to propose a science basedapproach for performing the related cumulative risk assessment (CRA). The project
progressed through six project tasks, culminating in this report (Task 6). The report is
broadly structured according to the project tasks but flexibility was used to allow the report
content to be arranged suitably for the scientific content.
Task 1 was to collect and scrutinize the relevant scientific literature. A search strategy
was refined to identify relevant experimental mixture studies and the literature was compiled
into a Microsoft Access database, termed the CRADIS (Cumulative Risk Assessment of
Dissimilarly acting chemicals) database (sections 4-7). The CRADIS database accompanies
this report and contains 173 experimental mixture studies (section 8). The search was
performed in Web of Knowledge (Thomson Reuters) to give the best coverage of the
literature. Experimental studies were classified and analysed within the database. All
chemicals included in the experimental studies were indexed allowing an overview of the
types of chemicals being studied in the literature. A deep analysis of experimental design and
results was restricted to studies considered to have relevance to two issues: 1) low dose issues
and 2) the issue of chemical dissimilarity. The literature identified in this task served as a
basis for Task 2.
Task 2 was to summarize and assess the state of the science. Relevant mixture studies
were analysed and revealed that the majority of the literature deals with studies comprising
only a few components, often only two components were studied (section 9). The literature
relating to low dose issues and to chemical dissimilarity was reviewed in detail (section 10).
In this task definitions of dissimilar mode of action were compiled and compared (section 11)
and legislation relating to cumulative risk assessment was reviewed (section 12). The
available assessment concepts for mixture toxicity, including independent action (IA) and
dose addition (DA) , were reviewed and this included considerations of the theoretical
background, applicability, data requirements and empirical evidence for each concept
(section 13). A thorough analysis of the quantitative difference between DA and IA
predictions was considered pivotal to this project (section 13.4) and revealed that prediction
might be expected to differ by less than one order of magnitude, even for mixtures with a
high number of components. Finally, cumulative risk assessment approaches based on the
available mixture concepts were reviewed (section 13.6) and the data requirements set in
legislation were analysed (section 13.9). The state of the science summary arising from this
task underpins the work in the subsequent Tasks 3, 4 and 5.
Task 3 was to propose scientific criteria for establishing cumulative assessment groups
of pesticides and other types of chemicals when dissimilar mode of action is a relevant
mechanism leading to a common effect, in order to perform dietary cumulative risk
assessment for regulatory purposes. The review of literature relevant to this task was
included in the report section for task 2, with a review of the definitions that are relevant to
dissimilar modes of action. In this section a physiologically based approach to grouping was
reviewed. The development and use of grouping criteria are integral to the approach that is
proposed in task 5, therefore proposals relevant to this are detailed in that task (section 15).
Supporting publications 2012:EN-232
3
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Task 4 was to assess the available approaches and methods for risk assessment of
mixtures of pesticides and other chemicals in or on foods showing dissimilar mode of
action. The approaches discussed by various bodies (US EPA, EFSA, WHO/IPCS) are
described and analysed (section 14). The use of tiered approaches in cumulative risk
assessment were considered in detail (section 14.3) and integrated into the final proposal.
Task 5 was to propose a science-based approach for performing cumulative risk
assessment of chemicals in food acting through dissimilar modes of action. An approach
is proposed that unifies the assessment of similarly and dissimilarly acting chemicals based
on pragmatic use of assessment approaches derived from the concept of DA (section 15). The
approach incorporates a tiered framework in line with other proposals such as that developed
by IPCS (2009). The approach suggests to abandon distinctions according to (presumed)
modes of action of chemicals at lower tiers of the analysis and to assess all the chemicals that
occur together in the exposure scenario under investigation. At higher tiers, and only if the
risks identified at lower tiers are deemed unacceptable, should chemicals be grouped together
according to their ability to evoke a common adverse outcome. Features of the proposed
approach are illustrated in three case studies (section 15.8).
Conclusions. The overall conclusion of this project is that it is feasible and justified to utilise
CRA methods and tiered framework analyses originally developed for similarly acting
mixtures also for combinations of dissimilarly acting chemicals. There can be one unified
approach for dealing with mixtures in regulatory practice, irrespective of (often presumed)
modes of action.
Recommendations. It is recommended to use a tiered framework analysis with CRA derived
from DA also for the assessment of mixtures of dissimilarly acting chemicals. At lower tiers
of the analysis, all chemicals deemed relevant for the exposure scenario under investigation
should be assessed, irrespective of their presumed modes of action. At higher tiers, when the
risk estimates at lower tiers are deemed unacceptable, chemicals known not to contribute to a
relevant common adverse outcome can be excluded from the analysis. By way of further
refining the analysis, criteria for the grouping of chemicals into common assessment groups
based on their capability of affecting a common adverse endpoint, should be applied.
Supporting publications 2012:EN-232
4
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table of Contents
Abstract ..................................................................................................................................................... 2 Summary ................................................................................................................................................... 3 Table of Contents ...................................................................................................................................... 5 Background ............................................................................................................................................. 10 Terms of reference .................................................................................................................................. 12 Acknowledgements ................................................................................................................................. 15 Introduction and Objectives .................................................................................................................... 16 Introduction ............................................................................................................................................. 16 1. Project scope .............................................................................................................................. 16 2. Guiding questions ...................................................................................................................... 16 3. Project tasks ............................................................................................................................... 17 Objectives ............................................................................................................................................... 17 Materials and Methods ............................................................................................................................ 19 4. Data collection ........................................................................................................................... 19 5. Systematic literature search ....................................................................................................... 20 5.1. Description of the search strategy that was adopted .............................................................. 20 5.2. Development and implementation of the literature search strategy....................................... 22 5.2.1. Aim of the search strategy ................................................................................................ 22 5.2.2. Search strategy details ....................................................................................................... 22 5.2.2.1. Search sources .......................................................................................................... 22 5.2.2.2. Document types: primary articles and reviews ......................................................... 23 5.2.2.3. Use of MeSH terms .................................................................................................. 23 5.2.2.4. Filtering by journal or research field ........................................................................ 23 5.2.2.5. Synonym selection, preliminary searches ................................................................ 23 5.2.2.6. Reasons for differences in number of results between PubMed and WoK .............. 24 5.2.2.7. Duplicate detection and removal .............................................................................. 24 5.2.2.8. Relevance of results to the topic of this report ......................................................... 25 5.2.2.9. Import of results into the project database (CRADIS) ............................................. 25 6. Ad hoc approaches ..................................................................................................................... 30 7. The CRADIS database ............................................................................................................... 31 7.1. Initial analysis (linking to chemicals) .................................................................................... 31 7.2. Mixture analysis .................................................................................................................... 31 7.3. Deep analysis ......................................................................................................................... 32 Results ..................................................................................................................................................... 33 TASK 1: information collection ............................................................................................................. 33 8. CRADIS database, current status and contents .......................................................................... 33 TASK2: state of the science summary .................................................................................................... 34 9. Results of CRADIS analysis ...................................................................................................... 34 9.1. Number of mixture components in experimental studies ...................................................... 34 9.2. Publication date of included studies ...................................................................................... 36 9.3. Chemicals included in experimental mixture studies ........................................................... 37 10. Primary literature ....................................................................................................................... 38 10.1. Experimental studies relevant to chemical dissimilarity ....................................................... 38 10.1.1. Non-chemical stressors ..................................................................................................... 38 10.2. Experimental studies relevant to low dose ............................................................................ 41 10.3. Experimental studies relevant to both dissimilar chemicals and low dose (“dual relevance”)43 Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
10.3.1. Analysis of chemicals and mode/mechanism of action information in dual relevance
studies 49 10.4. Method development ............................................................................................................. 53 10.4.1. Risk assessment approaches .............................................................................................. 53 10.4.2. Mathematical models ........................................................................................................ 53 11. Definitions of dissimilar mode of action.................................................................................... 56 11.1. UK Committee on Toxicity (2002) ....................................................................................... 56 11.2. European Food Safety Authority (2008) ............................................................................... 56 11.3. US Environmental Protection Agency (2002) ....................................................................... 57 11.4. Agency for Toxic Substances and Disease Registry (2004) .................................................. 57 11.5. WHO/IPCS (2009) ................................................................................................................ 57 11.6. Norwegian Scientific Committee for Food Safety (2008) ..................................................... 57 12. Legislation ................................................................................................................................. 60 12.1. Consideration of possible cumulative risk in EU pesticide regulations ................................ 60 12.1.1. Legal background .............................................................................................................. 60 12.1.2. Consideration of cumulative risk in EU legislation .......................................................... 60 12.2. Refusal of the authorisation of PPP ....................................................................................... 61 13. Assessment concepts for mixture toxicity and empirical evidence for their validity ................ 63 13.1. Independent action (IA) ......................................................................................................... 64 13.1.1. Applicability of IA to mixtures composed of agents with dissimilar modes of action ..... 64 13.1.2. Data requirements for using IA ......................................................................................... 65 13.1.3. Under IA, when is a mixture risk acceptable?................................................................... 65 13.1.4. Correlation assumptions in IA and their consequences..................................................... 66 13.1.5. Empirical evidence for IA ................................................................................................. 67 13.2. Dose addition (DA) ............................................................................................................... 68 13.2.1. Data requirements for using DA ....................................................................................... 69 13.2.2. Under DA, when is a mixture risk acceptable? ................................................................. 69 13.3. The role of mode and mechanism of action information in the choice between IA and DA 71 13.3.1. Definitions of mode and mechanism of action .................................................................. 71 13.3.2. Use of empirical evidence to infer similarity or dissimilarity ........................................... 73 13.3.3. Conclusions ....................................................................................................................... 74 13.4. Quantitative differences between DA and IA predictions .................................................... 75 13.4.1. Problem formulation ......................................................................................................... 75 13.4.2. Empirical evidence ............................................................................................................ 76 13.4.3. Limiting factors and resulting maximal differences .......................................................... 76 13.4.3.1. The number of mixture components (n) ................................................................... 78 13.4.3.2. The dose ratio of mixture components ..................................................................... 78 13.4.3.3. The slopes of dose response curves .......................................................................... 79 13.4.3.4. The effect of threshold assumptions ......................................................................... 82 13.4.4. Sample calculations and simulations ................................................................................. 84 13.4.4.1. Deterministic simulations of extreme prediction differences ................................... 85 13.4.4.2. Probabilistic simulations of distributions of prediction differences ......................... 87 13.4.5. Conclusions ....................................................................................................................... 90 13.5. Toxicological interactions ..................................................................................................... 91 13.6. Approaches to cumulative risk assessment methods ............................................................. 93 13.6.1. Approaches based on dose addition (DA) ......................................................................... 93 13.6.1.1. Hazard Index ............................................................................................................ 93 13.6.1.2. Toxic Unit Summation ............................................................................................. 94 13.6.1.3. Point of Departure Index .......................................................................................... 94 13.6.1.4. Relative Potency Factors .......................................................................................... 94 Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.6.1.5. Toxic Equivalency Factors ....................................................................................... 95 13.6.1.6. Data requirements and applicability of the cumulative risk assessment methods .... 95 13.6.2. Approaches based on independent action (IA).................................................................. 96 13.6.3. Use of the TTC in CRA approaches.................................................................................. 97 13.6.3.1. The TTC concept ...................................................................................................... 97 13.6.3.2. Use of the TTC in cumulative risk assessment ......................................................... 98 13.7. Cumulative risk from sub-ADI levels.................................................................................. 100 13.7.1. Extrapolation from NOAEL or Benchmark Dose to ADI ............................................... 101 13.7.1.1. 10 x 10 Assessment Factors.................................................................................... 101 13.7.1.2. Interspecies Assessment Factor .............................................................................. 101 13.7.1.3. Intraspecies Assessment Factor .............................................................................. 102 13.7.1.4. Continuous Responses ............................................................................................ 103 13.7.1.5. Quantal (Dichotomous) Responses ........................................................................ 106 13.7.1.6. Conclusion .............................................................................................................. 109 13.8. The issue of non-adverse effects at levels below ADIs ....................................................... 110 13.8.1. Effects below a critical effect size ................................................................................... 110 13.8.2. Issues arising from the dichotomization of continuous effect variables ......................... 111 13.8.3. Precursor Effects ............................................................................................................. 111 13.8.4. Biochemical Alterations .................................................................................................. 113 13.8.5. Conclusions ..................................................................................................................... 116 13.9. Data requirements in legislation and availability of such data ............................................ 117 13.9.1. Toxicology data available to regulators .......................................................................... 117 13.9.1.1. Legal requirements for the provision of toxicity data for food improvement agents118 13.9.1.2. Legal requirements for the provision of toxicity data for substances for nutritional
purposes 120 13.9.1.3. Legal requirements for the provision of toxicity data for food contact materials .. 121 13.9.1.4. Legal requirements for the provision of toxicity data for novel foods and novel
food ingredients ....................................................................................................................... 123 13.9.1.5. Legal requirements for the provision of toxicity data for residues ......................... 123 13.9.1.6. Legal requirements for the provision of toxicity data for contaminants................. 124 13.9.1.7. Legal requirements for the provision of toxicity data for undesirable substances in
animal nutrition – ..................................................................................................................... 124 13.9.1.8. Legal requirements for the provision of toxicity data for feed additives ............... 125 13.9.1.9. Summary and conclusions ...................................................................................... 126 13.9.2. Exposure data available to regulators.............................................................................. 127 13.9.2.1. Legal requirements for the provision of exposure data for pesticide residues ....... 128 13.9.2.2. Legal requirements for the provision of exposure data for substances in live
animals and products of animal origin ..................................................................................... 130 13.9.2.3. Legal requirements for the provision of exposure data for food improvement
agents
130 13.9.2.4. Legal requirements for the provision of exposure data for food supplements ....... 131 13.9.2.5. Legal requirements for the provision of exposure data for food contact materials 131 13.9.2.6. Legal requirements for the provision of exposure data for food contaminants ...... 131 13.9.2.7. Summary and conclusions ...................................................................................... 131 13.10. Cumulative exposure assessment (CEA) ............................................................................. 133 TASK 3: Grouping criteria ................................................................................................................... 134 TASK 4: Assess approaches to CRA for dissimilarly acting chemicals, evaluate the dose additivity
approach ................................................................................................................................................ 135 14. Existing approaches to CRA for dissimilarly acting chemicals ............................................... 135 14.1. Commonalities of approaches.............................................................................................. 138 Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
14.2. Practical implications for CRA of chemicals with diverse uses and properties and present in
foods 138 14.3. Assessment frameworks and tiering .................................................................................... 139 TASK5: propose an approach to CRA (dissimilarity) .......................................................................... 142 15. A tiered framework analysis for combinations of dissimilarly acting chemicals .................... 142 15.1. A unified approach to cumulative risk assessment for similarly and dissimilarly acting
chemicals .......................................................................................................................................... 143 15.2. Principal assumptions, simplifications and requirements .................................................... 143 15.3. Rules for step-wise refinements as the analysis moves to the next higher tier .................... 145 15.4. Elements of cumulative risk assessment in a tiered framework analysis ............................ 145 15.5. Framework analysis: initial considerations ......................................................................... 145 15.5.1. Tier 0 ............................................................................................................................... 146 15.5.2. Tier 1 ............................................................................................................................... 146 15.5.3. Tier 2 ............................................................................................................................... 147 15.5.4. Tier 3 ............................................................................................................................... 147 15.6. When should the risk characterisation step result in risk management measures? .............. 148 15.7. Criteria for the grouping of chemicals in CRA at higher tiers............................................. 148 15.8. Case studies ......................................................................................................................... 151 15.8.1. Case study 1: pesticides and contaminants in lettuce ...................................................... 152 15.8.1.1. Assessment Scenario A .......................................................................................... 153 15.8.1.2. Assessment Scenario B ........................................................................................... 153 15.8.1.3. Assessment Scenario C ........................................................................................... 155 15.8.1.4. Higher Tier Assessments ........................................................................................ 155 15.8.1.5. Conclusions ............................................................................................................ 156 15.8.2. Case study 2: Mycotoxins in food commodities ............................................................. 166 15.8.3. Case study 3: dietary exposure to pesticides ................................................................... 172 15.8.3.1. Description of dataset ............................................................................................. 172 15.8.3.2. Tier 1: HI analysis (all ADIs) ................................................................................. 172 15.8.3.3. Interpretation of HI and HQ values ........................................................................ 173 15.8.3.4. HI analysis using TTC values (pseudo tier 0 analysis) .......................................... 174 15.8.3.5. PODI (NOAELs) .................................................................................................... 175 15.8.3.6. Tier 2 (consideration of effects) ............................................................................. 176 15.8.3.7. Breakdown of HI analysis using PPDB health issues ............................................ 177 15.8.3.8. Tier 3 (group according to known or plausible toxicological independence) ........ 178 15.8.3.9. Conclusions ............................................................................................................ 178 Conclusions and Recommendations ..................................................................................................... 195 Conclusions ........................................................................................................................................... 195 16. Conclusions .............................................................................................................................. 195 17. Identification of data gaps ........................................................................................................ 196 Recommendations ................................................................................................................................. 197 References ............................................................................................................................................. 198 18. References cited in this report.................................................................................................. 198 18.1. Legislation and regulations cited (sections 12 and 13.9) ..................................................... 206 18.2. References included in the CRADIS database .................................................................... 210 Appendices............................................................................................................................................ 230 Appendix A ........................................................................................................................................... 230 Appendix B ........................................................................................................................................... 230 Appendix C ........................................................................................................................................... 230 Glossary / Abbreviations....................................................................................................................... 231 Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Supporting publications 2012:EN-232
9
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Background
As provided by EFSA, section I.1.2 BACKGROUND INFORMATION, Tender specifications
for project CFT/EFSA/PPR/2010/02)
Regulation (EC) No. 396/2005 on Maximum Residue Levels (MRLs) of pesticides in or on
food and feed emphasises the importance “of carrying out further work to develop a
methodology to take into account cumulative and synergistic effects of pesticides” for dietary
risk assessment.
As there are currently no harmonised or internationally agreed methodologies to assess risks
from exposure to more than one compound, EFSA is developing methodologies to assess
risks arising from exposures to more than one active compound in food.
On 28/29 November 2006, EFSA started working on cumulative risk assessment of pesticides
by organising a colloquium on “Cumulative risk assessment of pesticides to human health:
the way forward”.
Based on the results from this international event, EFSA s Scientific Panel on Plant
Protection Products and their Residues (PPR Panel) elaborated an opinion “to evaluate the
suitability of existing methodologies and, if appropriate, identify new approaches to assess
cumulative and synergistic risk from pesticides to human health with a view to setting MRLs
for those pesticides within the framework of Regulation (EC) No. 336/2005” (EFSA, 2008).
Following the general opinion of the PPR Panel, a worked example was developed for a
group of triazole compounds and the results were reported in a separate opinion with
suggested additional refinements of the methodology (EFSA, 2009). In this opinion the PPR
Panel also proposed criteria for inclusion of compounds in a cumulative assessment group
(CAG), highlighting the possibility of different levels of refinement in a step-wise approach.
Before practical implementation of cumulative risk assessments for regulatory purposes, the
PPR Panel is currently working on a third opinion identifying pesticides that can, based on
their structure and effects, be grouped together for cumulative risk assessment. In the
preparatory phase of this opinion, the PPR Panel outsourced the information collection aimed
at establishing a database with cumulative assessment groups.
As an additional progress in the hazard assessment for cumulative risk assessment, the state
of the science on combined actions of pesticides and chemicals with dissimilar mode of
action, including endocrine disruptors, should be investigated. Evidence in recently published
scientific literature shows that certain endocrine disruptors show a dose additive effect even
if they do not share the same primary molecular target (Kortenkamp, 2007 and the papers
there reviewed; Christiansen et al. 2008; Moretto, 2008; Anonymous, 2009; Jacobsen et al,
2010; Reffstrup et al, 2010). Considering these results, the criterion for establishment of
cumulative assessment groups of pesticides and chemicals in or on foods with dissimilar
modes of action leading to a common effect, should be further elaborated. This should apply
in particular, but not only, to pesticides interacting with or affecting the endocrine system. It
has also been postulated that this type of combined toxicity can be addressed by the doseaddition approach (EFSA, 2008; University of London, 2009).
Supporting publications 2012:EN-232
10
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
References:
Anonymous, 2009. Expert workshop on combination effect of chemicals, Hornbaeck,
Denmark, organized by Danish Ministry of the Environment and the Danish Environmental
Protection Agency.
Christiansen S., Scholze M., Axelstad M., Boberg J., Kortenkamp A., Hass U., 2008.
Combined exposure to anti-androgens causes markedly increased frequencies of hypospadias
in the rat. International Journal of Andrology, 31, 241-248.
EFSA (European Food Safety Authority), 2008. Scientific Opinion of the Panel on Plant
Protection Products and their Residues (PPR Panel) on a request from the EFSA evaluate the
suitability of existing methodologies and, if appropriate, the identification of new approaches
to assess cumulative and synergistic risks from pesticides to human health with a view to set
MRLs for those pesticides in the frame of Regulation (EC) 396/2005. The EFSA Journal
(2008) 704, 1-84.
EFSA (European Food Safety Authority), 2009. Scientific Opinion of the Panel on Plant
Protection Products and their Residues (PPR Panel) on risk assessment for a selected group
of pesticides from the triazole group to test possible methodologies to assess cumulative
effects from exposure through food from these pesticides on human health. The AFSA
Journal (2009) 7(9), 1167.
Jacobsen P. R., Christiansen S., Boberg J., Nellemann C., Hass U., 2010. Combined exposure
to endocrine disrupting pesticides impairs parturition, causes pup mortality and affects sexual
differentiation in rats. International Journal of Andrology, 33, 434-442.
Kortenkamp, A., 2007. Ten years of mixing cocktails: a review of combination effects of
endocrine-disrupting chemicals. Environmental Health Perspectives, 115, 98-105.
Moretto A., 2008. Exposure to multiple chemicals, when and how to assess the risk from
pesticide residues in food. Trends in Food Science & Technology, 19, S56-S63.
Refstrup T. K., Larsen C. J., Meyer O., 2010. Risk assessment of mixtures of pesticides,
Current approaches and future strategies. Regulatory toxicology and Pharmacology 56, 174192.
University of London, 22 December 2009. State of the Art Report on mixture toxicity, Final
Report.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Terms of reference
As provided by EFSA, tender specification to project CFT/EFSA/PPR/2010/0, sections I.1.3
and I.1.4
PURPOSE OF THE ASSIGNMENT
Overall objective(s):
The overall objective of the contract resulting from the present procurement procedure is to
provide scientific information on different aspects of combined actions of chemicals in food
acting through dissimilar modes of action and to define criteria regarding the elaboration of
cumulative assessment groups of pesticides which do not necessarily share a common
mechanism or mode of action. The review should be applied to chemicals present in food in
general and not restricted to pesticides in order to obtain wider information. The information
will be used by the EFSA PPR Panel to further refine the hazard assessment for dietary
cumulative risk assessment.
Specific objectives:
The specific objectives of the contract resulting from the present procurement procedure are
as follows:
•
Information gathering on the state of the science on combined actions of chemicals
with dissimilar mode of action, including endocrine disruptors. The starting point for
literature search should be the “State of the Art Report on Mixture Toxicity”
published by the European Commission (EC) in 2009, available on the EC
Environment website. The contractor should also consider the report on Risk
Assessment of Mixtures of Pesticides and Similar Substances (Committee on
Toxicity, UK, 2002) and the Opinion of the Scientific Steering Committee of the
Norwegian Scientific Committee for Food Safety, Combined Toxic Effects of
Multiple Chemical Exposures (VKM Norway, 2008) to avoid any duplication of
works.
•
Elaboration of general criteria for establishment of cumulative assessment groups of
pesticides and chemicals in or on food acting through dissimilar modes of action
leading to a common effect. This should apply in particular, but not only, to pesticides
interacting with or affecting the endocrine system.
•
Proposal for science-based approach for performing cumulative risk assessment of
chemicals in food acting through dissimilar modes of action, including endocrine
active substances.
•
Identification of research needs to improve the understanding of the relevance of
independent action for mixture toxicity.
SCOPE OF THE WORK, EXPECTED OUTCOMES AND DELIVERABLES, TIMELINE
AND PAYMENTS
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The contractor selected as a result of this tendering procedure is expected to carry out the
following tasks:
Collect and scrutinize all recently published information from scientific literature relevant to
combined actions of chemicals showing dissimilar mode of action and not covered by the EC
report. Particular emphasis will be given to data at low dose (below the NOAEL).
•
Provide a comprehensive summary and assessment of the collected information in
view of understanding
o the nature and site of actions,
o possible targets,
o interactions with different subsystems of the affected organisms and the
different types of the possible combined actions when the dissimilar mode of
action is to be considered.
During this activity the references valid for low dose mixtures in EC, COT and VKM
reports should be identified. The identified studies in these reports should be deeply
analysed if
•
-the compounds have common phenomenological end points,
•
-the tested concentrations were appropriate to trigger observable effects,
•
-the study protocols had really the aptitude to validate the independent action
theory,
•
-the observed effects were analysed considering the need of different
approaches to different types of dose-response data (continuous, quantal or
ordinal).
•
The toxicological endpoints and type of effects relevant for the cumulative risk
assessment based on a dissimilar mode of action should be identified in particular, but not
only, for chemicals interacting with or affecting the endocrine system.
•
Propose scientific criteria for establishing cumulative assessment groups of pesticides
and other types of chemicals when dissimilar mode of action is a relevant mechanism leading
to a common effect, in order to perform dietary cumulative risk assessment for regulatory
purposes.
•
Assess the available approaches and methods for risk assessment of mixtures of
pesticides and other chemicals in or on foods showing dissimilar mode of action. In the report
on mixture toxicity (EC, 2009) the dose addition approach was considered to be probably
conservative enough in most of the cases. Scrutinizing the collected information the
contractor should identify cases, if any, where this approach would be underconservative.
•
Propose a science-based approach for performing cumulative risk assessment of
chemicals in food acting through dissimilar modes of action.
Supporting publications 2012:EN-232
13
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
•
Provide a structured report of the requested activities.
Supporting publications 2012:EN-232
14
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Acknowledgements
This contract was awarded by EFSA to:
The School of Pharmacy, University of London (ULSOP)
Contract/grant title: Investigation of the state of the science on combined actions of
chemicals in food through dissimilar modes of action and proposal for science-based
approach for performing related cumulative risk assessment
Contract/grant number: CT/EFSA/PPR/2010/04
Supporting publications 2012:EN-232
15
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Introduction and Objectives
INTRODUCTION
This is the final report for the project titled “Investigation of the state of the science on
combined actions of chemicals in food through dissimilar modes of action and proposal for
science-based approach for performing related cumulative risk assessment”
(CFT/EFSA/PPR/2010/02).
1.
Project scope
The project was specified so as to have relevance for:
•
Dietary cumulative risk assessment; which could include for example pesticides,
contaminants, additives, contact materials and toxins
•
Regulatory purposes; which encompasses regulatory requirements and legal
background
•
Current practice; for example having consideration for the EFSA practice on CRA for
similarly acting chemicals, as shown by the 2008 Opinion (EFSA 2008b) and the
2009 triazole case study (EFSA 2009c).
2.
Guiding questions
In addition to the specified project tasks and objectives, we have identified a number of
guiding questions which we have used to direct and structure our work on the project and this
report. The guiding questions were:
•
What theories and definitions are available or applied to considerations of dissimilar
action in the context of cumulative risk assessment?
•
Is there an approach to mixture assessment based on the independent action (IA)
approach?
•
In 2009, there was no available mammalian example of an experimental situation
when IA gave a different effect prediction to dose addition (DA) and was also
accurate (EC 2009). Has this position changed in scientific literature since 2009?
•
Is there an experimental example in the scientific literature when IA provided the
more conservative prediction, compared to DA?
•
When using the IA concept, when is risk considered acceptable?
Supporting publications 2012:EN-232
16
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
3.
Project tasks
The project was organized into 6 tasks, the tasks and their relationship to each specific
objective (SO) of the project are shown in Figure 1, and are listed here:
Task 1 was to collect and scrutinize the relevant scientific literature. The major output of this
task is a Microsoft Access database containing the relevant literature.
Task 2 was to summarize and assess the state of the science. Particular attention has been
paid to definitions of dissimilar mode of action and to legislation relating to cumulative risk
assessment. The state of the science summary underpins the subsequent Tasks 3, 4 and 5;
which feed directly into Task 6 (Final Report).
Task 3 was to propose scientific criteria for establishing cumulative assessment groups of
pesticides and other types of chemicals when dissimilar mode of action is a relevant
mechanism leading to a common effect, in order to perform dietary cumulative risk
assessment for regulatory purposes.
Task 4 was to assess the available approaches and methods for risk assessment of mixtures of
pesticides and other chemicals in or on foods showing dissimilar mode of action. In the report
on mixture toxicity (EC, 2009) the dose addition approach was considered to be probably
conservative enough in most of the cases. Scrutinizing the collected information the
contractor should identify cases, if any, where this approach would be underconservative.
Task 5 was to propose a science-based approach for performing cumulative risk assessment
of chemicals in food acting through dissimilar modes of action.
Task 6 was to provide a structured report of the requested activities
OBJECTIVES
The specific objectives listed in the project tender specifications from EFSA are listed in
Figure 1, which also shows how the objectives are related to the project tasks.
Supporting publications 2012:EN-232
17
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
SO1. Information gathering
Task 1: collect recent studies, focus on dissimilar mode of action, low dose issues
Task 2: summarise and assess collected data
SO2. Elaboration of general criteria Task 3: propose grouping criteria
Task 4: assess available approaches for dissimilar mode of action, assess appropriateness of dose additivity approach
SO3. Proposal for science‐based approach Task 5: propose approach for cumulative risk assessment of chemicals acting through dissimilar modes of action
SO4. Identification of research needs Task 6: provide report on requested activities
Figure 1: Relationship between project tasks and the specific objective (SO) of the
project
Supporting publications 2012:EN-232
18
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Materials and Methods
4.
Data collection
Data collection (Task 1) was performed in a systematic fashion, and was organised around a
literature database (Microsoft Access), referred to as the CRADIS database, which stands for
Cumulative Risk Assessment of DISsimilarly acting chemicals. The literature database was
populated with two strategies: firstly, a systematic literature search was carried out with the
aim of identifying peer-reviewed literature relevant to the application of independent action
(IA) as a concept in mixture toxicology; secondly, ad hoc approaches were used to identify
documents pertinent to the project but not identified in the systematic search. In particular,
studies relating to mixtures of chemicals considered to be ‘dissimilar’, and studies relating to
‘low-dose’ issues were sought. The data collection strategy is illustrated in Figure 2.
In the remainder of this methods section the systematic search is described in section 5, ad
hoc approaches in section 6 and the resulting literature database is described in section 7.
Relevant articles from key reports
Literature search focused on independent action, and synonyms
COT, 2002; VKM, 2008; ULSOP, 2009
Ad hoc searches
PubMed
&
Web
Of
Knowledge
Author names
Keywords
Expert input
Selected journals
CRADIS
Bibliographic data
Abstract
Screen for relevance, document decision
Article annotation
Link chemicals
Deep analysis
Mixtures, dissimilarity, low‐dose
Obtain full text
Figure 2: Schematic diagram showing the data collection strategy
Supporting publications 2012:EN-232
19
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
5.
Systematic literature search
5.1.
Description of the search strategy that was adopted
This section describes the important details of the search strategy that was adopted. The
following section (section 5.2) provides more detail about the development and refinement of
the strategy.
The search was performed in Web of Knowledge (Thompson Reuters). A cut-off date of
March 2011 was agreed at the First Coordination Meeting (Parma, 17th January 2011) and the
search was updated in April and May 2011 to ensure coverage of literature published, and
indexed, up to the cut-off date.
The search phrases used are listed in Box 1.
Box 1: search strategy The following quoted phrases were used to query Web of Knowledge (Thomson Reuters, wok.mimas.ac.uk): “independent action” “independent joint action” “effect summation” “effect addition” “response addition” “response summation” “bliss independence” “simple similar action” “effect multiplication”* “response additivity”** Target databases included: Medline (1950‐present), Web of Science (1970‐present), BIOSIS Previews (1969‐present) All phrases were joined with an ‘OR’. Accepted document types were: article, review *note that searches showed all results (6) for this term were OFF domain.
** Inclusion of this term added 18 articles, of which 6 were considered relevant. The most recent article was published in
2009, preceded by 2 published in 2005; suggesting that ‘response additivity’ is not a commonly used term in current
parlance.
The search had a deliberately wide focus to reduce the chance of omitting relevant studies,
consequently a large number of articles (approximately 1,150) were identified. The abstract
of each article was obtained and scanned to assess the relevance of the article to the project.
Articles were classified as follows:
•
OFF domain: articles that are not relevant, and that were returned due to the use of
search phrases in other contexts than mixture toxicology
Supporting publications 2012:EN-232
20
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
•
ON domain: articles that had relevance to the project. These results were further
classified as
o Trivial: articles that had some relevance but that were of minor use to the
project aims
o Experimental: articles that described an experimental study, usually of
chemical mixtures, in which mixture toxicity concepts, including independent
action, were employed.
o Method development: articles that developed methodology for dealing with
mixture, or frameworks and approaches to cumulative risk assessment
o Reviews
The classification was performed in a secondary Microsoft Access database, and a report
from that database, showing the classification of each article, is included as Appendix A.
Around 79% of the search results were classified as ‘off domain’, this was due to the
deliberately wide focus of the search. The use of an Access database allowed this sorting of
many irrelevant hits to be done efficiently and with full documentation of the process
(Appendix A).
The remaining documents, around 250, were considered potentially relevant to the project,
however around 17% were classified as having minor relevance, and were not analysed
further. This produced 203 documents for further consideration, of which 80% dealt with
experimental studies, 9% dealt with method development and 10% were relevant reviews.
Supporting publications 2012:EN-232
21
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
5.2.
Development and implementation of the literature search strategy
This section provides further detail of the development and refinement of the search strategy
stated in section 5.1.
5.2.1.
Aim of the search strategy
The aim was to identify a suitable search strategy that could be used prior to a manual
inspection and filtering of the results, ultimately producing a relevant set of documents
without major omissions.
For this purpose it is acceptable if the search initially includes false positives (i.e. results
include articles that are not relevant) but not if there are many false negatives (i.e. results do
not include relevant articles). This is because manual checking will allow for the elimination
of false positives, but cannot compensate for documents that are missing due to being false
negatives. False negatives can be dealt with by expert input of domain specific knowledge or
by serendipitous/ad hoc browsing through the literature, for example looking at other articles
in the same journal issue as a relevant article can identify other relevant articles that were not
returned by the search process used. That aim of refining the search strategy was to identify
which search terms should be included to give the best chance of retrieving documents
relevant to the independent action model used in mixture toxicology. This includes
identification of useful synonyms, and suitable combinations thereof, for example are
relevant documents found by one important keyword or is the use of several synonyms
necessary.
The use of synonyms is not consistent in the field of mixture toxicity, therefore some terms
may not be true synonyms, in that some authors may use them to refer to another concept,
however they must be included in the search strategy so as not to discount the instances
where they are used to refer to the concept of interest here. For those cases when the
synonym is phrase is used but is not used as a synonym then manual inspection will identify
them as off-domain.
5.2.2.
Search strategy details
5.2.2.1. Search sources
Two sources were evaluated: PubMed (public MEDLINE) which is publically available from
the US National Library of Medicine and Web of Knowledge, which is available through
subscription (Thomson Reuters).
Searches were carried out using the web interfaces provided, for PubMed (www.pubmed.org)
the main search box on the home page was used. For WoK (www.wok.mimas.ac.uk) the
search box for the topic field was used.
Supporting publications 2012:EN-232
22
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
5.2.2.2. Document types: primary articles and reviews
Both data sources, PubMed and WoK, tag articles according to the document type, for
example primary article, review, conference proceeding etc. We considered the possible use
of these tags to limit the search strategy.
In PubMed the use of “Review[pt]”, where [pt] indicates publication type, in the search string
should limit the results to review articles, however this approach was not adopted since visual
inspection of the results showed that some articles tagged in this way were in fact primary
research papers. In WoK, selection of document type = “REVIEW”, also included primary
research papers.
In WoK, a greater number of document types are present, for example conference
proceedings and book chapters, that are not generally present in PubMed. It was desirable to
remove these types of results since they increase the apparent number of results without
being likely to contain relevant information, for example, although conference proceedings
can be an indicator of ongoing research, the level of detail provided is not usually sufficient
for thorough analysis, and the item may not have undergone full peer review.
Consequently, tags were not used in PubMed and the counts shown in the table below are for
all results and, in WoK, document types of “ARTICLE” and “REVIEW” were
included/selected but “MEETING”, “ABSTRACT” and other types were excluded.
5.2.2.3. Use of MeSH terms
MeSH terms can be a useful way to separate articles that include a search phrase being used
in comm. on usage from those that use the phrase as specialist terminology. A preliminary
search for important terms (“concentration addition”, “dose addition” and “independent
action”) indicated that the MeSH vocabulary lacks these terms for mixture concepts and that
therefore database curators (PubMed, MEDLINE and WoK) will not have been able to use
MeSH terms to tag articles as relating to specific mixture concepts. MeSH terms were
therefore not used as part of the search strategy.
5.2.2.4. Filtering by journal or research field
Certain databases provide a field for indicating the area of research, and this may be a way to
limit the number of irrelevant documents that are found in a search. However preliminary
investigation of this option suggested that the process is not transparent (for example, it is not
clear how journals and/or articles are assigned to the research field), and it is not easy to
identify journals that definitely will or will not contain ‘relevant’ documents. Consequently
this option was not utilised since it might have led to the omission of relevant documents.
5.2.2.5. Synonym selection, preliminary searches
To identify relevant synonyms and the extent to which searches in PubMed and WoK
produce overlapping results, a series of searches were carried out using selected search
strings in both sources and the number of results (“hits”) was compared. The results of
searches for “independent action” and synonyms are shown in Table 1. For the purposes of
Supporting publications 2012:EN-232
23
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
comparison, similar searches to those listed above were carried for concentration addition
and synonyms thereof. These results are shown in Table 2.
This analysis indicated that few synonyms were used redundantly, and that all synonyms
should be used to give the best chance of the search identifying relevant documents.
5.2.2.6. Reasons for differences in number of results between PubMed and WoK
Initial searches indicated that WoK and PubMed searches typically produce different
numbers of results. Generally, WoK produced more results than PubMed. Visual inspection
suggests that some of these ‘additional’ results are indeed relevant, i.e. they are not a
consequence of including irrelevant results, and that they should not be omitted. A potential
reason for these differences is the journals that are indexed, in particular PubMed is directed
towards the medical literature, although it is by no means limited to medical journals.
PubMed have also adopted an updating model that allows journals to submit entries which
are almost immediately available online with minimal checking by PubMed/NLM, in contrast
WoK does their own curation of database records, for example indexing the references that
are cited by an article, and this process takes time. The additional curation by WoK provides
additional features at the cost of immediacy.
WoK was adopted as the main source of searches in this project because of its greater
coverage, and because it also includes MEDLINE results.
5.2.2.7. Duplicate detection and removal
The presence of duplicate results increases the redundancy of a database, and can have an
impact of the amount of work involved in processing a set of database records. We therefore
considered whether the search strategy resulted in duplicate records. We have not noticed the
presence of duplicate records in PubMed, however WoK does include duplicate records. We
were able to remove some of these by use of the duplicate identification algorithm included
in the Reference Manager software that was used to process the results of the search strategy.
The remainder were identified by sorting the database in alphabetical order by title and visual
scanning to identify neighbouring identical titles and author lists.
One explanation of duplicates could be the presence of an advance or ‘e’ publication as well
as the final authoritative print version, which often has a substantially later date of
appearance.
Around 30 documents were found to be duplicated, and these duplicate entries were removed.
Supporting publications 2012:EN-232
24
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
5.2.2.8. Relevance of results to the topic of this report
After duplicate scanning, titles were read to decide if they were relevant to the topic: “IA as a
mathematical concept for the joint action of chemicals, usually in toxicology”
For this analysis articles were loaded into an access database for classification of their
relevance (OFF domain, ON domain (trivial, experimental study, method development,
review)). Classification was made on the basis of title, abstract (if available) and publication
date (if an abstract was unavailable, articles before 2000 were not usually pursued unless the
title/date indicated high relevance). Database name: classification of IA search
results.accdb
Table 3 shows the result of this analysis. Out of 1153 results from the literature search, 203
were considered relevant; this is shown in the table as “ON domain” and not “TRIVIAL”.
32 of these results were not found in PubMed (so 171 were in PubMed, and were linked to
their PMIDs); so around 16% of the articles derived from WoK only, and using WoK in
addition to PubMed identified 18% extra articles. Table xx lists the journals that articles
came from that were only retrieved via WoK.
5.2.2.9. Import of results into the project database (CRADIS)
The 203 articles that were relevant (ON domain and NOT trivial) were then imported to the
CRADIS database for further analysis. Articles with a PMID (PubMed identifier) were
imported automatically. A Visual Basic code was written that used the PMID to download
citation details and abstract from the PubMed server in the tagged MEDLINE format and
parsed the results into the appropriate CRADIS database fields), articles that did not have
PMIDs were manually entered into the CRADIS database.
26 of the identified articles had already been entered into CRADIS and 32 articles did not
have PMIDs (they were unique to Web of Knowledge, see Table 4); therefore 142 articles
were loaded automatically using their PMID and 32 articles were manually loaded.
The CRADIS database is fully described in section 7.
Supporting publications 2012:EN-232
25
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 1: results of searches in PubMed and WoK for ‘independent action’ and
synonyms thereof
PubMed
“independent action”
“simple dissimilar action”
Simple
AND dissimilar
AND action
“simple independent action”
492
Not found**
11 (1)
Web of
Knowledge
Hits (number “on
domain”*)***
31397/
28075
768/672
2
29/23 (6)
7 (6)
7/7 (6)
“Independent joint action”
“Response addition”
“Response summation”
“response addition”
AND “response summation”
“Effect addition”
“Effect summation”
“effect addition”
AND “effect summation”
“response addition”
OR “response summation”
OR “effect addition”
OR “effect summation”
12 (11)
42
16
0
23/22 (19)
166/156
45/41
1
19
10
0
293/272
19/17
0
WoK: 262 if language =English
87
522/485
PM: results show there is no overlap
There is only one paper with more
than one of these phrases in it; a
search within these results for
‘independent action’ = 20 hits
“independent action”
OR “independent joint action”
OR “effect summation”
OR “effect addition”
OR “response addition”
OR “response summation”
“bliss independence”
“effect multiplication”
581
1297/
1163
28
0
55/30
6 (0)
Search string
Independent action
Hits (number
“on domain”*)
21782
Notes
PM: phrase used once in 2011, then
1998; focus of 6 articles was
epidemiological and mathematical
WoK: phrase last used in 1998
PubMed and WoK results differ by
one paper each
WoK: All 6 results were OFFDOMAIN
Not a useful term
*question used to decide relevance was “did the paper deal with the mathematical concept used in mixture toxicology?”
**search returned no results. Search engine response was “quoted phrase not found” (PubMed) or “Search Error: Invalid
query. Please check syntax” (WoK).
*** shows aaa/bbb (n), when aaa = count of all results, bbb= count of results for ARTICLES and REVIEW document types,
n=number of documents that were relevant.
All database searches were performed on 6.4.11
Supporting publications 2012:EN-232
26
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 2: results of searches in PubMed and WoK for ‘concentration addition’ and
synonyms thereof
Search string
“Dose addition”
“dose additivity”
“dose addition” AND
“dose additivity”
“Dose additive”
“Concentration
addition”
“Concentration
additivity”
“Concentration
addition” AND
“Concentration
additivity”
“concentration
additive”
“dose addition” AND
concentration addition”
“simple similar action”
“dose addition” OR
“dose additivity” OR
“concentration
addition” OR
concentration
additivity” OR “simple
similar action”
“dose addition” OR
“dose additivity” OR
“dose additive” OR
“concentration
addition” OR
“concentration
additivity” OR
“concentration
additive” OR “simple
similar action”
PubMed
Hits (n)
78
35
4
Notes
Web of Knowledge
Hits (n)***
Notes
131/107
52/42
6/6
Shows 6 papers ‘overlap’
56
187
73/56
515/479
11
22/18
0
1
21
45/44
0
1 paper overlaps
PM: Note that this
result shows there
is NO overlap
7
310
14/12
720/645
361
801/714
*question used to decide relevance was “did the paper deal with the mathematical concept used in mixture toxicology?”
**search returned no results. Search engine response was “quoted phrase not found” (PubMed) or “Search Error: Invalid
query. Please check syntax” (WoK).
*** shows aaa/bbb, when aaa = count of all results, bbb= count of results for ARTICLES and REVIEW document types
All database searches were performed on 6.4.11
Supporting publications 2012:EN-232
27
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 3: results of classifying search results
Notes on definition
All results
OFF domain
ON domain
TRIVIAL
EXPERIMENTAL STUDY
METHOD DEVELOPMENT
REVIEW
Supporting publications 2012:EN-232
1. Did not match topic, e.g. glucose
independent action of drug A,
response summation and psychology,
independent action of two genes (no
mathematical concept implied)
2. there was only a title (no abstract)
and the title did not have enough
information to deem the article
relevant
3. article in a foreign language (n=1)
1. Title only, appears relevant from
the information in the title. If the title
appeared very relevant then efforts
were made to find the abstract or full
text, especially if the document was
recent (post 2000)
N
1153
907
43
163
19
21
28
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 4: list of journals from which articles came that were unique to WoK searches
Journal title
Environmental Toxicology And Chemistry
Asian Journal of Ecotoxicology
Human And Ecological Risk Assessment
Biometrics
Journal Of Applied Ecology
Canadian Journal Of Fisheries And Aquatic Sciences
Chinese Science Bulletin
Continental Shelf Research
Crop Protection
Crop Research (Hisar)
Fresenius Environmental Bulletin
Green Chemistry
Journal Of Economic Entomology
Journal Of Environmental Quality
Phytopathology
Scandinavian Journal Of Statistics
Umweltwissenschaften Und Schadstoff-Forschung
Weed Research
Weed Technology
Supporting publications 2012:EN-232
Number of articles
7
4
3
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
29
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
6.
Ad hoc approaches
Ad hoc approaches were used to allow flexibility in identifying literature that was not found
in the systematic literature search but that was identified, for example, by expert input.
Approaches included:
•
identification of relevant documents from the core reports: COT, 2002; VKM, 2008
and EC, 2009
•
targeted literature searches for key authors
•
searches of specific journals, issues
•
expert input
•
citation searches (Web of Knowledge, Thomson Reuters)
We evaluated the utility of using citation searches, in which articles citing a key paper are
located, rather than a search strategy based on keywords. However we found that this
approach generated large numbers of results, was dependent on the ‘seed’ key paper selected
and relied on complete, comprehensive citation of prior papers by authors – which is not
usually done. Dependence on the ‘seed’ paper can be reduced by using several or many
seeds, however this increases the size of the output above manageable levels. Consequently
citation searches were used as part of the ad hoc strategy, not used systematically.
Supporting publications 2012:EN-232
30
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
7.
The CRADIS database
Results from the search strategies described above were compiled into a Microsoft Access
database, referred to as the CRADIS (Cumulative Risk Assessment of DISsimilarly acting
chemicals) database. The database accompanies the Final Report and the filename is
“CRADIS.accdb”.
The database contains two main tables, one containing bibliographic information and analysis
of articles (“ARTICLES”) and the other containing chemical information (“CHEMICALS”).
Each table has a unique ID field that is used for linking between the tables. The ARTICLES
table is linked to a sub table called “MIXTURES” which details a mixture experiment. The
tables are presented to the database user through a series of forms, starting with a menu form
that allows the user to access the database in three ways:
1. from an articles perspective, when the user starts from an article or list of articles and can
then ‘click-through’ to linked information in the MIXTURES and CHEMICALS tables
2. from a chemicals perspective, when the user start from a chemical of interest and can click
through to linked information in the ARTICLES and MIXTURES tables
3. from a list of database reports. The user is presented with a list of preformatted reports that
are built from the database contents.
Initially all articles identified were listed in the database, then bibliographic, abstract and
linking information (linking to PubMed through PubMed identifiers (PMID), linking to full
text sources through Digital object identifiers (DOI)) was added.
Articles were then classified as to their area of relevance to the key issues for this project:
mixture studies, primarily those studies with relevance to chemical dissimilarity, and lowdose. Full text was sourced for relevant articles. Relevant documents were then annotated
further, and subjected to deeper analysis as described next.
7.1.
Initial analysis (linking to chemicals)
Links to related chemicals were made by connecting entries on the ARTICLES table to
entries on the CHEMICALS table. The use of linking allows chemical information, such as
abbreviations, full names, mw, and ID numbers to be entered once only.
7.2.
Mixture analysis
For all studies, the number of mixture components, n, was documented. This result is shown
on the reports page of the CRADIS database in a PivotTable showing the size of n in all of
the mixture studies. When the study included multiple mixtures, the highest n was entered.
For mixtures including nonchemical stressors, the nonchemical stressor was included in the
calculation of n. Selected studies, those with relevance to both dissimilarity and low dose
Supporting publications 2012:EN-232
31
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
issues, were analysed in more detail and parameters such as mixture design, prediction
models used, results observed, analyses used etc were documented, see section 7.3.
7.3.
Deep analysis
Those articles deemed relevant to both of the key issues for the project, low dose and
dissimilarity, were subject to a deeper analysis of the mixture design and results. This is
documented on the ‘mixture’ and ‘deep analysis’ tabs in the CRADIS database.
Supporting publications 2012:EN-232
32
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Results
TASK 1: INFORMATION COLLECTION
Task 1 entailed the design, creation and population of an annotated literature database
CRADIS (Microsoft Access). The database forms the basis of the scientific summary (Task
2) and provides source material for subsequent tasks.
8.
CRADIS database, current status and contents
The CRADIS database contains 266 articles, including primary literature, reviews and
scientific reports. These are listed in the reference list (section 18).
•
Virtually all of the articles have abstracts, and full text was sourced for 168 articles.
•
All articles have been classified as to their area of relevance to the project.
•
Chemicals tested in 173 mixture studies have been indexed to allow direct retrieval of
all the studies relevant to a given chemical.
•
In doing so, 631 chemicals have been indexed and linked to the experimental studies
that included them.
•
Mixture studies identified as relevant to both chemical dissimilarity and to low-dose
issues have been subjected to a deep analysis, as specified in the project proposal and
the Tender Specifications.
Supporting publications 2012:EN-232
33
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TASK2: STATE OF THE SCIENCE SUMMARY
This section presents a summary of the state of the science, which is the primary output of
Task 2. The summary focuses on an analysis of the published mixtures studies that were
compiled into the CRADIS database (see Task 1) and a review of the critical issues in the
field. This review served as the basis for the work performed in fulfilment of project tasks 3,
4 and 5.
9.
Results of CRADIS analysis
9.1.
Number of mixture components in experimental studies
Figure 3 presents a pie chart showing the number of components examined in each of the
experimental mixture studies contained within the CRADIS database. This analysis shows
that the majority (52%) of studies examined binary mixtures (2 components), and that less
than 25% of all the studies examined mixtures with more than six components. The highest
number of components studied was 33 (Hermens et al. 1985).
This focus on binary mixtures most likely reflects the intention of developing mixture
concepts rather than being a practical attempt to screen chemicals, since it is evident that the
systematic examination of binary mixtures of all possible chemical combinations would be
unfeasible. Mixture studies with few components have a low ability to determine whether
DA and IA are better predictors of observed data because the two predictions are more likely
to be close to each other as the number of components decreases (see section 13.4).
Supporting publications 2012:EN-232
34
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 3: pie chart showing the number of components investigated in experimental
studies of mixtures
Supporting publications 2012:EN-232
35
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
9.2.
Publication date of included studies
The bar chart below indicates the publication year of the articles and reviews included in the
CRADIS database. The upward trend reflects both the growing number of publications in
this field and the intentional focus on recent literature in this state of the science summary.
Figure 4: bar chart showing publication year of articles included in CRADIS
Supporting publications 2012:EN-232
36
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
9.3.
Chemicals included in experimental mixture studies
In addition to the analysis required by the tender specifications, the chemicals tested in all
mixture studies (173) for which details were available were indexed and linked to the
experimental studies in which they were tested. The indexed chemicals are listed in Appendix
B, and the frequency with which chemicals were tested is indicated graphically in a Wordle
diagram below.
Figure 5: Wordle diagram showing a visual representation of the chemicals examined,
and the frequency of their inclusion, in experimental studies
Figure shows the chemicals tested in mixture studies in a Wordle plot (www.wordle.net) in
which word size is proportional to the number of studies that the chemical was tested in. For
orientation to the relationship between word size and inclusion, copper was included in 12
studies whilst bisphenol A was included in 6. The full list of chemicals is provided in the
CRADIS database and in Appendix B of this report.
Supporting publications 2012:EN-232
37
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
10.
Primary literature
The CRADIS database contains 174 experimental studies of mixtures, of which 94 were
classified as having relevance to dissimilar chemicals and 26 were classified as having
relevance to low dose issues. 13 studies were classified as relevant to both dissimilar
chemicals and low dose issues. Each of these literature groups is now summarized.
10.1.
Experimental studies relevant to chemical dissimilarity
We identified and listed the criteria used by authors of mixture studies relevant to
dissimilarity to assign similarity or dissimilarity. Note that at this stage the term
‘dissimilarity’, and to some extent by implication also ‘similarity’, encompasses most
possible usages of the term (s).
These results are found in the “MixGroupingCriteria” field in the “Articles” table of the
CRADIS database. The main approaches were:
•
Mode/mechanism of action/toxicity. In most cases a framework for establishing the
mode/mechanism of action was not stated, and we expect that different authors
performed this analysis in different ways. This may be unsurprising since it has been
often noted that there is no current framework or established vocabulary for
describing mechanism of toxicity.
•
Chemical grouping, for example as metals, pesticides, herbicides, insecticides and
ionic liquids. We note that terms such as ‘pesticide’, ‘herbicides’, and ‘insecticides’
refer to a common use of chemicals with potentially diverse structures and probably
should not be considered as groups of structurally related chemicals unless further
clarification is provided, for example ‘neonicotinoid insecticides’.
•
Opposing actions, mixtures of chemicals with directly opposing (cytochrome p450
inducers and inhibitors (Scott and Hodson 2008)) or indirectly opposing (mixtures of
estrogens and antiandrogens (Eustache et al. 2009)) actions.
Approximately half of the studies used some version of mode/mechanism of action whilst
around one third used an approach based on some kind of chemical grouping.
10.1.1.
Non-chemical stressors
In addition to studies of combinations of chemicals, seven studies that included chemicals
with a nonchemical stressor were identified and considered as being relevant to dissimilar
action. These studies are analysed in Table 5. All of the studies were ecotoxicological in
focus. All of the combinations studied were binary and included combinations of one
chemical (including carbaryl, carbendazim, cadmium, fluoranthene, imidacloprid, nickel,
nonylphenol or potassium dichromate) and one nonchemical stressor (including low/high
temperature; low dissolved oxygen levels, low soil moisture, perceived predation risk/threat,
parasitism and low photon flux).
Supporting publications 2012:EN-232
38
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
All of the studies compared experimental results with a prediction based on IA, whilst around
half of the studies also compared to a prediction based on DA. In most cases IA was
considered as the reference model, as opposed to the situation for studies of chemicals only
when DA is often considered as the reference model, even if IA is also frequently evaluated.
The inclusion of nonchemical stressors in CRA may therefore change the appropriate
position of IA in CRA approaches, as described in later project tasks. The inclusion of
nonchemical stressors in CRA has been called for, but has not been done to date (NRC 2009).
Supporting publications 2012:EN-232
39
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 5: analysis of mixture studies that included nonchemical stressors
Study
chemical
Nonchemical stressor
Organism
(Ferreira et al. 2010)
Nickel
Low/high temperature;
low dissolved oxygen levels
Daphnia magna Straus
(water flea)
(Jensen et al. 2009)
Nonylphenol
Low/high temperature
Dendrobaena octaedra
(earthworm)
IA
(Long et al. 2009)
Fluoranthene
low soil moisture
Lumbricus rubellus
(earthworm)
IA
(Pestana et al. 2009)
Imidacloprid
perceived predation risk
IA, DA
(Coors and De
Meester 2008)
(Ferreira et al. 2008)
Carbaryl
predation threat;
parasitism
low dissolved oxygen levels
Sericostoma vittatum
(caddisfly);
Chironomus riparius
(midge)
Daphnia magna (water
flea)
Daphnia magna Straus
(water flea)
Scenedesmus
subspicatus (green alga)
IA, DA, effect
summation
(Cleuvers et al. 2002)
Cadmium,
carbendazim
Potassium
dichromate
Supporting publications 2012:EN-232
low photon flux
Mixture concepts
used
IA, DA
IA
IA, DA
Conclusions of authors
Deviations from IA were noted (both
synergy and antagonism) whilst DA could
be a poor fit to the observed data.
IA was used as the reference model, the
authors observed strong synergism with
nonylphenol and high but not low
temperature.
IA “provided a good description of the
combined stressor data…and was the most
parsimonious model describing joint effect”
The majority of parameters showed no
deviation from IA or DA
IA was found to provide useful, quantitative
predictions of effect
Observed adequate prediction by IA in some
cases, underprediction by both IA and DA in
other cases, and also observed antagonism
from IA.
Combination effects “could be calculated
well only by IA”
40
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out exclusively by the author(s) in the context of a contract between the European Food
Safety Authority and the author(s), awarded following a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety Authority is subject. It may
not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the
authors.
Combined actions of chemicals in food through dissimilar modes of action
10.2.
Experimental studies relevant to low dose
We compiled the definition of low dose used by 27 studies, and this information can be found
in the “LowDoseDefinition” field of the “Articles” table in the CRADIS database and is also
shown in Table 6.
Ten studies used no observed effect concentrations (NOEC), sometimes with an estimation
that this was equivalent to an EC01, and a further seven used ‘less than NOEC’ or a fraction
of NOEC (one third, one quarter or one tenth). These studies are listed in the following table.
Two studies used a set effect level, of EC05 (Deng et al. 2007) or EC10 (Merino-Garcia et al.
2003), and one study used 0.04 times the LC50 (Hermens et al. 1985).
Other studies used the MRL (Wade et al. 2002) or ADI (Ito et al. 1995) and two studies used
a measure of human levels, human serum concentrations (van Meeuwen et al. 2007) or
background human daily intake (Crofton et al. 2005). One study used a set concentration,
1mg/kg/day (Eustache et al. 2009).
This analysis shows that various definitions of low dose are in use in the literature, and that
their meanings are sufficiently different to complicate comparison of studies described as low
dose without further information and analysis.
Table 6: Low dose definitions used in the primary literature
Low dose definition
Authors
Pub. year
NOEC
Backhaus T, Scholze M, Grimme LH
2000
NOEC
2002
NOEC
Walter H, Consolaro F, Gramatica P, Scholze M,
Altenburger R
Kunz PY, Fent K
NOEC, EC01
Kunz PY, Fent K
2009
NOEL, EC01
Manzo S, Buono S, Cremisini C
2010
chemical doses that had no
observable effect when tested alone
Christiansen S, Scholze M, Axelstad M, Boberg J,
Kortenkamp A, Hass U
2008
NOEC (=EC01)
Silva E, Rajapakse N, Kortenkamp A
2002
sub-effective concentrations,
expected to be similar to EC01
Belden JB, Lydy MJ
2006
ED01
van Meeuwen JA, van den Berg M, Sanderson JT, Verhoef
A, Piersma AH
Wang Z, Chen J, Huang L, Wang Y, Cai X, Qiao X, Dong
Y
2007
Broderius SJ, Kahl MD, Elonen GE, Hammermeister DE,
Hoglund MD
Broerse M, van Gestel CA
2005
NOEC, EC01
EC01
2006
2009
Less than, or fraction of, NOEC
<NOEC
<NOEC
Supporting publications 2012:EN-232
2010
41
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
<NOEC, EC01
Faust M, Altenburger R, Backhaus T, Blanck H, Boedeker
W, Gramatica P, Hamer V, Scholze M, Vighi M, Grimme
LH
2003
<NOEC, EC01
Faust M, Altenburger R, Backhaus T, Blanck H, Boedeker
W, Gramatica P, Hamer V, Scholze M, Vighi M, Grimme
LH
2001
1/10 NOAEL
Jonker D, Woutersen RA, van Bladeren PJ, Til HP, Feron
VJ
Jonker D, Woutersen RA, van Bladeren PJ, Til HP, Feron
VJ
Groten JP, Schoen ED, van Bladeren PJ, Kuper CF, van
Zorge JA, Feron VJ
1990
EC05
Deng Fc;Liu Ss;Liu Hl;Mo Ly;
2007
EC10
fraction of LC50
Merino-Garcia D, Kusk KO, Christensen ER
2003
0.04 x LC50
Hermens J, Leeuwangh P, Musch A
1985
Wade MG, Foster WG, Younglai EV, McMahon A,
Leingartner K, Yagminas A, Blakey D, Fournier M,
Desaulniers D, Hughes CL
Ito N, Hasegawa R, Imaida K, Kurata Y, Hagiwara A,
Shirai T
2002
van Meeuwen JA, Ter Burg W, Piersma AH, van den Berg
M, Sanderson JT
Crofton KM, Craft ES, Hedge JM, Gennings C, Simmons
JE, Carchman RA, Carter WH Jr, DeVito MJ
2007
Eustache F, Mondon F, Canivenc-Lavier MC, Lesaffre C,
Fulla Y, Berges R, Cravedi JP, Vaiman D, Auger J
2009
not clearly defined
Pavlaki MD, Pereira R, Loureiro S, Soares AM
2011
"low concentration-response range"
Petersen K, Tollefsen KE
2011
1/4 NOAEL
MinimumOAEL (MOAEL),
NOAEL, 1/3 NOAEL
ECx
1993
1997
MRL, ADI
MRL, TDI, NOEL
ADI, 100xADI
1995
Human levels
human serum concentrations
background human daily intake
2005
set concentration
1 mg/kg/day
Not clearly defined
Supporting publications 2012:EN-232
42
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
10.3.
Experimental studies relevant to both dissimilar chemicals and low dose (“dual
relevance”)
14 studies were considered to have relevance to both of the main topics of this project,
dissimilar chemicals and low dose. These studies were subject to a full analysis of their
experimental content in the CRADIS database. Four of the studies were published in the
review period of this report (2008 – March 2011) and are reviewed below. A review of the
experimental literature prior to this period can be found in the State of the Art Report on
Mixture Toxicity (EC 2009). Key features of all fourteen studies, including those prior to the
review period, are summarised in Table 7, and chemical and mode/mechanism information
from all fourteen studies was analysed, see section 10.3.1.
Our analysis included comparing each experimental study to five quality criteria that have
been previously applied to mixture studies (EC 2009;Kortenkamp et al. 2007) This analysis is
included in Table 7. The quality criteria are:
A: toxicity of individual mixture components was experimentally determined under identical
conditions as the mixture;
B: stability of test concentrations under test conditions was checked by analytical methods
(does not apply to animal experiments with direct dosing);
C: uncertainty of experimentally determined effects, effect concentrations, or effective doses
was estimated by statistical methods;
D: uncertainty of mixture toxicity predictions was estimated by statistical methods; and
E: no observed effect concentrations (NOECs) or NOELs were determined for every
individual substance, and individual concentrations or doses resulting in the given joint effect
were demonstrated to be at or below these NOECs or NOELs, or insignificance of individual
effects was demonstrated by other statistical approaches (Kortenkamp et al. 2007).
We note that the recent experimental studies published since the 2009 EC report have not
significantly altered the state of the mixture toxicology field however they are reviewed
briefly here because they were identified by the exhaustive search strategy that was adopted
for the project. An important question for this project was whether there is empirical
evidence of relevance to the use of IA in risk assessment, this question was not answered by
these recent studies and the state of the science on this issue is reviewed separately in section
13.1.5.
•
Chronic dietary exposure to a low-dose mixture of genistein and vinclozolin
modifies the reproductive axis, testis transcriptome, and fertility (Eustache et al.
2009).
Eustache et al examined chronic exposure to low doses of endocrine disruptors and
administered binary mixtures of a phytoestrogen, genistein, and an antiandrogen, vinclozolin
Supporting publications 2012:EN-232
43
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
to male rats from conception to adulthood at doses of 1, 10 and 30 mg/kg/day. We cannot
analyse this study in detail because the authors did not use mixture toxicity concepts to
formulate quantitative effect predictions, which is an important quality criteria for mixture
studies (Kortenkamp et al. 2007).
•
Integrated fuzzy concentration addition-independent action (IFCA-IA) model
outperforms two-stage prediction (TSP) for predicting mixture toxicity (Wang et
al. 2009).
Wang et al. studied 12 industrial organic chemicals (Benzene, toluene, chlorobenzene,
phenol, aniline, nitrobenzene, 3-nitrotoluene, 2,4-Dichlorophenol, hydroquinone, 3nitrochlorobenzene, 4-nitrophenol, 2,4,6-trichlorophenol) with four different modes of toxic
action (nonpolar narcotic, polar narcotic, pro-electrophile, oxidative phosphorylation
uncoupling) with the aim of comparing the ability of an integrated fuzzy concentration
addition-independent action (IFCA-IA) model with the two stage prediction (TSP) model. In
the TSP model chemicals are grouped by their model of action, then firstly the effect of each
group is calculated using DA (referred to by Wang at al as concentration addition (CA)), and
secondly these group effects are cumulated using IA. Conversely, the IFCA-IA model does
not use MOA information and instead uses fuzzy logic to calculate weight coefficients for
DA and IA from molecule structure descriptors. Wang et al. exposed Vibrio fischeri to three
mixtures: (1) in ratio of individual EC01 concentrations; (2) in ratio of individual EC50
concentrations; (3) in ratio of equimolar concentrations. The IFCA-IA model must be trained
on a data set, and Wang et al. used the experimental results of their EC01 mixture, see next,
as the training set. Wang et al. found that the IFCA-IA model outperformed the TSP model
with prediction errors for the EC01, EC50 and equimolar mixtures of 0.3, 6 and 0.6% for
IFCA-IA and 2.8, 19 and 24% for TSP.
The requirement for training data means that the IFCA-IA approach is likely to have practical
obstacles to application in human risk assessment when it is likely that most mixtures of
interest will not have been tested experimentally. This study is also reviewed in section
10.4.2, concerning recent method developments.
•
Mixture effects of nickel and chlorpyrifos on Folsomia candida (Collembola)
explained from development of toxicity in time (Broerse and van Gestel 2010).
Broerse and van Gestel used IA and DA to study the effects of a seven week exposure to
binary mixtures of nickel and chlorpyrifos on Folsomia candida , a soil-dwelling Arthropod
(collembola). The authors found that chlorpyrifos toxicity exhibited an extremely steep doseresponse curve that meant the data was unsuitable for use in IA or DA models, however they
found that careful monitoring of toxicity allowed the determination of mixture effects, and
reported that, even at exposure levels below the No Effect Concentration, chlorpyrifos was
able to reduce the toxicity of nickel.
This study indicated the difficulty of confounding toxicity in mixture studies, but had few
wider implications for the field.
•
Effects of binary mixtures on the life traits of Daphnia magna (Pavlaki et al.
2011).
Supporting publications 2012:EN-232
44
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Pavlaki et al. compared the toxicity of binary mixtures of imidacloprid with either thiacloprid
or nickel in Daphnia magna. Imidacloprid and thiacloprid are both neonicotinoid insecticides,
for which a common mode of action may be likely. For the mixture of two insecticides, the
authors observed synergy (neonatal number) and conformance to DA (body length). For the
mixture of insecticide with metal, the authors observed conformance to IA (neonatal number)
and a dose dependent effect on body length with synergy at low dose and antagonism at
higher doses. This study provides an example of when IA can predict the mixture effect
(neonatal number), however in this case the experimental data was also compatible with the
DA prediction.
Supporting publications 2012:EN-232
45
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 7: Summary and comparison of studies with dual relevance to the project.
Title of paper
Author list (Pub.
Date)
Notes
Joint toxicity of mixtures
of groups of organic
aquatic pollutants to the
guppy (Poecilia
reticulata).
4-week oral toxicity study
of a combination of eight
chemicals in rats:
comparison with the
toxicity of the individual
compounds.
Effect of ingestion of 20
pesticides in combination
at acceptable daily intake
levels on rat liver
carcinogenesis.
Subacute toxicity of a
mixture of nine chemicals
in rats: detecting
interactive effects with a
fractionated two-level
factorial design.
Effects of subchronic
exposure to a complex
mixture of persistent
contaminants in male rats:
systemic, immune, and
reproductive effects.
Hermens J,
Leeuwangh P,
Musch A (1985)
Analysed in table
6.4 (EC, 2009)
Jonker D,
Woutersen RA, van
Bladeren PJ, Til
HP, Feron VJ
(1990)
Mixture toxicity of
priority pollutants at no
observed effect
concentrations (NOECs).
Joint toxicity of similarly
and dissimilarly acting
chemicals to Daphnia
Low dose
definition used
Quality
criteria
fulfilled
A
0.04 x LC50
Analysed in table
6.5 (EC, 2009)
ACE
1/10 NOAEL
Ito N, Hasegawa R,
Imaida K, Kurata
Y, Hagiwara A,
Shirai T (1995)
Analysed in table
6.5 (EC, 2009)
C
ADI, 100xADI
Groten JP, Schoen
ED, van Bladeren
PJ, Kuper CF, van
Zorge JA, Feron VJ
(1997)
Analysed in table
6.5 (EC, 2009)
ABCE
MinimumOAEL
(MOAEL),
NOAEL, 1/3
NOAEL
Wade MG, Foster
WG, Younglai EV,
McMahon A,
Leingartner K,
Yagminas A,
Blakey D, Fournier
M, Desaulniers D,
Hughes CL (2002)
Walter H,
Consolaro F,
Gramatica P,
Scholze M,
Altenburger R
(2002)
Analysed in table
6.5 (EC, 2009)
effects: systemic,
immune,
reproductive
CE
MRL, TDI,
NOEL
Analysed in table
6.4 (EC, 2009)
"These findings
strongly indicate,
that the modes of
action of mixture
components are
indeed dissimilar in
a way, that is
sufficient to predict
the mixture toxicity
with the concept of
independent
action."
ABCE
NOEC
AC
EC10
Merino-Garcia D,
Kusk KO,
Christensen ER
Supporting publications 2012:EN-232
46
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
magna at different
response levels.
(2003)
Joint algal toxicity of 16
dissimilarly acting
chemicals is predictable
by the concept of
independent action.
Faust M,
Altenburger R,
Backhaus T, Blanck
H, Boedeker W,
Gramatica P,
Hamer V, Scholze
M, Vighi M,
Grimme LH (2003)
Broderius SJ, Kahl
MD, Elonen GE,
Hammermeister
DE, Hoglund MD
(2005)
A comparison of the lethal
and sublethal toxicity of
organic chemical mixtures
to the fathead minnow
(Pimephales promelas).
Thyroid-hormonedisrupting chemicals:
evidence for dosedependent additivity or
synergism.
Integrated fuzzy
concentration additionindependent action
(IFCA-IA) model
outperforms two-stage
prediction (TSP) for
predicting mixture
toxicity.
Chronic dietary exposure
to a low-dose mixture of
genistein and vinclozolin
modifies the reproductive
axis, testis transcriptome,
and fertility.
Mixture effects of nickel
and chlorpyrifos on
Folsomia candida
(Collembola) explained
Analysed in table
6.4 (EC, 2009)
ABCD
E
<NOEC, EC01
Effects: lethality,
sublethality (32d
growth)
30 individual
components were
tested, and several
mixtures were
tested: the largest
mixture was 12
components but all
had the same mode
of toxic action. The
largest mixture of
chemicals with a
different MTOA w
ABCD
<NOEC
AC
background
human daily
intake
AC
EC01
Crofton KM, Craft
ES, Hedge JM,
Gennings C,
Simmons JE,
Carchman RA,
Carter WH Jr,
DeVito MJ (2005)
Wang Z, Chen J,
Huang L, Wang Y,
Cai X, Qiao X,
Dong Y (2009)
Eustache F,
Mondon F,
Canivenc-Lavier
MC, Lesaffre C,
Fulla Y, Berges R,
Cravedi JP, Vaiman
D, Auger J (2009)
Broerse M, van
Gestel CA (2010)
Supporting publications 2012:EN-232
DISSIM: estrogen
plus antiandrogen
effects: standard
reproductive
toxicology end
points, testicular
mRNA expression
profiles
1 mg/kg/day
AB
<NOEC
47
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
from development of
toxicity in time.
Effects of binary mixtures
on the life traits of
Daphnia magna.
Pavlaki MD,
Pereira R, Loureiro
S, Soares AM
(2011)
Supporting publications 2012:EN-232
effects:
reproduction,
survival, body
length
AB
not clearly
defined
48
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
10.3.1. Analysis of chemicals and mode/mechanism of action information in dual
relevance studies
In fulfilment of Task 2, we have analysed the identified studies for information on the
chemicals tested in mixture studies, and the presence of mode of action information. The
chemicals tested in the 14 identified studies, see section 10.3 above, were listed and any
information regarding mode/mechanism of action provided by the authors of the papers, for
example as a basis for grouping, was compiled. 142 chemicals were included over the 14
studies, this information is listed in Appendix C.
Our analysis revealed that only limited information on mode of action was included in any
study, and that no consistent approach is used in the experimental literature to describe mode
of action information. Comparison of studies on the basis of mode of action is virtually
impossible.
64 chemicals had some mode/mechanism stated, although this includes 8 that were stated as
‘not known’. The information for 56 chemicals, with 26 modes/mechanisms is presented in
Table 8.
None of the chemicals were tested in more than two studies and only 17 chemicals were
included in two different studies. Of these 17 chemicals, 11 had no mode/mechanism
information in either study, and 6 had information from one study only. No chemicals had
mode/mechanism from both studies, and consequently the consistency of the schemes being
used to assign this information cannot be assessed. None of the studies explicitly stated a
scheme. The available information is presented in Table 9.
It should be noted that the studies included both mammalian toxicology and ecotoxicology
studies, therefore the modes/mechanisms listed derive from a wide range of species.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 8: list of 26 modes/mechanisms and the 56 associated chemicals
Mode/mechanism
Chemical name
amino acid biosynthesis
Carotenoid biosynthesis
DNA synthesis and function
enzyme inhibitor
inhibition of photosynthesis
Metsulfuron-methyl
Norflurazon
Nalidixic acid
ethyl-parathion
Tributyltin chloride
Triphenyltin chloride
Metazachlor
DTMAC
pentachloroethane
2,4-dimethylphenol
nonylamine
decylamine
Benzene
Toluene
chlorobenzene
Azaserine
4-nitrophenol
2,4,6-trichlorophenol
Paraquat dichloride
Terbuthylazine
Atrazine
Aniline
2,4-Dichlorophenol
Phenol
nitrobenzene
3-nitrotoluene
Aclonifen
hydroquinone
3-nitrochlorobenzene
Chloramphenicol
CCCP
Mixture study details:
First author, Pub. Year
Faust , 2003
Faust , 2003
Faust , 2003
Merino-Garcia , 2003
Walter , 2002
Walter , 2002
Faust , 2003
Faust , 2003
Broderius , 2005
Broderius , 2005
Merino-Garcia , 2003
Merino-Garcia , 2003
Wang , 2009
Wang , 2009
Wang , 2009
Faust , 2003
Wang , 2009
Wang , 2009
Faust , 2003
Faust , 2003
Walter , 2002
Wang , 2009
Wang , 2009
Wang , 2009
Wang , 2009
Wang , 2009
Faust , 2003
Wang , 2009
Wang , 2009
Faust , 2003
Faust , 2003
Hexachlorobutadiene
Fenfuram
Kresoxim-methyl
8-azaguanine
Metalaxyl
Triadimenol
2,4-dinitrophenol
Broderius , 2005
Faust , 2003
Faust , 2003
Faust , 2003
Faust , 2003
Faust , 2003
Broderius , 2005
lipid biosynthesis
Membrane functions
Narcosis I
Narcosis II
narcotic
Nonpolar narcotic
Nucleotide biosynthesis
Oxidative phosphorylation
uncoupling
Photosynthetic electron
transport
photosystem II inhibitor
Polar narcotic
porphyrin biosynthesis
Pro-electrophile
protein biosynthesis
Proton translocation and ATP
synthesis
reactive toxicant
Respiratory electron transport
RNA synthesis and function
steroid biosynthesis
uncoupler of oxidative
phosphorylation
Supporting publications 2012:EN-232
50
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table continued...
Upregulation of hepatic
catabolism of thyroid
hormones
TCDD
PCDD
TCDF
1-PCDF
4-PCDF
OCDF
PCB-28
PCB-52
PCB-77
PCB-101
PCB-105
PCB-126
PCB-138
PCB-153
PCB-156
PCB-169
PCB-180
PCB-118
Supporting publications 2012:EN-232
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
Crofton , 2005
51
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 9: chemicals tested in more than one “dual relevance” mixture study with
mode/mechanism information when stated
Chemical name
1,2,3-Trichlorobenzene
Mode/mechanism (if stated)
Study details: First author, Pub. Year
Wade , 2002
1,2,3-Trichlorobenzene
Hermens , 1985
1,2,3,4-tetrachlorobenzene
Wade , 2002
1,2,3,4-tetrachlorobenzene
Benzene
Hermens , 1985
Nonpolar narcotic
Wang , 2009
Benzene
Hermens , 1985
Pentachlorobenzene
Hermens , 1985
Pentachlorobenzene
Toluene
Wade , 2002
Nonpolar narcotic
Wang , 2009
Toluene
Hermens , 1985
2,4-Dichlorotoluene
Hermens , 1985
2,4-Dichlorotoluene
Hermens , 1985
Aniline
Hermens , 1985
Aniline
Polar narcotic
Wang , 2009
2,4-Dichlorophenol
Polar narcotic
Wang , 2009
2,4-Dichlorophenol
Phenol
Hermens , 1985
Polar narcotic
Wang , 2009
Phenol
Hermens , 1985
Mirex
Wade , 2002
Mirex
Jonker , 1990
Loperamide
Jonker , 1990
Loperamide
Groten , 1997
Stannous chloride
Jonker , 1990
Stannous chloride
Groten , 1997
Chlorpyrifos
Broerse , 2010
Chlorpyrifos
Ito , 1995
Endosulfan
Ito , 1995
Endosulfan
Wade , 2002
Cadmium chloride
Wade , 2002
Cadmium chloride
Groten , 1997
TCDD
Wade , 2002
TCDD
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
nickel
Pavlaki , 2011
nickel
Broerse , 2010
Supporting publications 2012:EN-232
52
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
10.4.
Method development
Some 30 articles in the CRADIS database deal with recent method developments in mixture
toxicology. We will now briefly summarize the studies organized by two topics: 1)
developments with potential relevance to risk assessment approaches and 2) development of
mathematical models for predicting mixture effects, especially mixed models of mixture
toxicity.
10.4.1. Risk assessment approaches
Method developments include one study that used Cramer classes and quantitative models of
uncertainty to model the effects of mixtures of migrant chemicals (Price et al. 2009). This
approach utilizes the existing IA and DA models with pragmatic approaches to data gaps.
Another study extended the approaches used in mixture toxicology to address chemical
exposures to the arena of multiple insect resistance genes (Wolt 2011), thus widening the
nature of mixtures for which regulatory approaches can be considered.
Of relevance to the use of mode of action to define similarity or dissimilarity is an IPCS
framework for assessing the relevance of a non-cancer mode of action that has been
published (Boobis et al. 2008), complementing a framework for mode of action in cancer
(Boobis et al. 2006). These frameworks are discussed in more detail in section 14.3.
10.4.2. Mathematical models
A number of articles have proposed new modelling or mathematical approaches to mixture
toxicity, often based on combinations or extensions of the existing IA and DA models. These
include:
•
A new log Kow-based model to predict the combined toxicities of antifouling
chemicals, which the authors found was not accurately predicted by IA or DA (Wang
et al. 2011).
•
A novel bio-concentration factor-based model to predict toxicity in the sea urchin
embryo-larval bioassay, developed after the authors found only weak correlation of
the observed toxicity with predictions using IA or DA (Xu et al. 2011).
•
A novel model of integrated concentration addition with independent action based on
multiple linear regression (ICIM, (Qin et al. 2011)). The authors consider that ICIM
may be useful when mixture components do not have a strictly similar or dissimilar
mode of action.
•
The Effect Residual Ratio (ERR) method (Wang et al. 2010) which can be used to
assess deviations from IA and DA predictions.
•
The MIXTOX model, developed in EU project ENV4-CT97-0507, was evaluated
using binary mixture studies of nickel, irgasan (an antibiotic) and diclofenac (an antiinflammatory)(Rudzok et al. 2010). The model incorporates the IA and DA models,
Supporting publications 2012:EN-232
53
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
and provides a framework for significance testing of deviations from reference
models (Jonker et al. 2005).
•
An Integrated Fuzzy Concentration Addition-Independent Action (IFCA-IA) model
which was evaluated in a study of 12 industrial organic chemicals with four different
modes of toxic action (Wang et al. 2009). The IFCA-IA model uses fuzzy logic to
derive weight coefficients from molecular structural descriptors, and uses these
coefficients to weigh the contribution of IA and DA to the model.
Two of these studies propose mixed models of mixture toxicity which combine the DA and
IA models and each is now examined in turn. Mixed models recognise that both the DA and
IA concepts can apply to mixture effects, but that realistic scenarios are unlikely to conform
strictly to either a pure DA or a pure IA prediction.
Integrated Fuzzy Concentration Addition-Independent Action (IFCA-IA) Model Outperforms
Two-Stage Prediction (TSP) For Predicting Mixture Toxicity (Wang et al. 2009)
Wang et al. proposed an Integrated fuzzy concentration addition-independent action (IFCAIA) model as an alternative to the two stage prediction (TSP) model. In the TSP model
chemicals are first grouped by their mode of action, then the effect of each group is
calculated using DA, and finally these group effects are cumulated using IA. Conversely, the
IFCA-IA model does not use MOA information and instead uses fuzzy logic to calculate
weight coefficients from molecule structure descriptors. The coefficients are used to set the
contribution of DA and IA to the overall model. The IFCA-IA model must be trained on an
experimental data set, which may not be available in the regulatory context. The chief benefit
of this model, that MOA information is not required, may not be sufficient to outweigh the
practical obstacle of q requirement of suitable training data. However this endeavour may
chime with other efforts to increase the use of read-across from well studied chemicals to less
studied chemicals, and may be a useful approach if it can be adapted from its current position
in experimental mixture studies into a practical tool in cumulative risk assessment.
A Novel Model Of Integrated Concentration Addition With Independent Action Based
On Multiple Linear Regression (ICIM) (Qin et al. 2011)
Qin et al proposed a model called Integrated Concentration addition with Independent action
based on Multiple linear regression (ICIM). ICIM uses multiple linear regression to derive a
relationship between observed mixture effects and the separate DA and IA predictions of
those effects. This approach, using multiple linear regression, contrasts with the use of fuzzy
logic based on molecular descriptors in models such as IFCA-IA. Qin et al found that ICIM
had a strong predictive power, and that it provided better predictions than either DA or IA.
However, like the IFCA-IA model discussed above, the ICIM model also requires training;
Qin et al used experimental results of mixtures from a ray design to predict the results of
Supporting publications 2012:EN-232
54
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
mixtures with an equipotent design. Consequently the ICIM may have similar limitations to
the IFCA-IA method and cannot be considered as a practical model for use in CRA.
To our knowledge, a practical CRA approach that allows a commonly used implementation
of DA, namely the Hazard index, to be combined with some implementation of IA is not in
use. This is a potentially important method gap.
The common feature of these mixed models is the use of DA and IA as the underlying
concepts. It follows logically that predictions made using these mixed models must fall
within the predictions made using pure DA and pure IA. If it is accepted that both data and
method gaps currently limit the use of a mixed model, then the use of this simplification, that
the mixed model prediction lies between DA and IA, may prove useful, especially if the
extent of the difference between DA and IA predictions is understood. The extent of, and
factors driving, the differences between DA and IA predictions is the topic of section 13.4.
Supporting publications 2012:EN-232
55
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
11.
Definitions of dissimilar mode of action
In fulfilment of work specified for Task 3, we have assembled the definitions used in major
reports or by regulatory organisations that relate to the issue of dissimilar modes of action. In
most cases the related terms were ‘simple similar action’ contrasted with ‘simple dissimilar
action’. We note that the definitions are used to refer to the same broad concepts, but differ in
the amount of detail that is specified within the definition.
The definitions provided for simple similar action and simple dissimilar action are
summarised in Table 10. The definitions are basically identical but differ in the amount of
detail given, e.g. whether they mention the target site or the relative potency.
11.1.
UK Committee on Toxicity (2002)
The UK Committee on Toxicity (COT) uses the term simple similar action in its report on
“Risk Assessment of Pesticides and Similar Substances” (COT 2002) for chemicals that “act
in the same way, by the same mechanism(s), and differ only in their potencies”. Dose
additivity is a consequence of simple similar action. In the case of simple dissimilar action
“it is assumed that the modes of action and possibly the nature and site of action differ
among the chemicals in the mixture which exert their individual effects, but do not modulate
the effect of other constituents of the mixture.” In the COT report dose addition is used as
synonym for simple similar action and independent action is used as synonym for simple
dissimilar action.
11.2.
European Food Safety Authority (2008)
The Panel on Plant Protection Products and their Residues (PPR) of the European Food
Safety Authority states that simple similar action “occurs when chemicals in a mixture act
in the same way, by the same mechanism/mode of action, and differ only in their
potencies”(EFSA 2008b). The effects of the mixture are equivalent to the sum of the potencycorrected doses of each component of the mixture. According to EFSA (2008) simple similar
action is a synonym for dose-addition (“dose-addition, also referred to as simple similar
action”). On the other hand simple dissimilar action is a synonym for response-addition
(“response-addition, also referred to as simple dissimilar action”) and “occurs where the
modes of action and possibly, but not necessarily, the nature and sites of toxic effects differ
between the chemicals in a mixture, and one chemical does not influence the toxicity of
another”. The effects of the mixture are equivalent to the combination of the effects of each
component of the mixture.
Supporting publications 2012:EN-232
56
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
11.3.
US Environmental Protection Agency (2002)
The US Environmental Protection Agency does not provide a final definition of simple
similar action in its “Guidance on cumulative risk assessment of pesticide chemicals that
have a common mechanism of toxicity” (EPA 2002). But they characterise “toxicologically
similar” chemicals as chemicals that “share a common toxic effect” or “act at the same target
site”. In contrast to the PPR/EFSA and the UK COT the US EPA does not use the term
“dose-addition” as a synonym for similar action. The US EPA recommends the assumption of
“dose-additivity” in the risk assessment procedure in case of chemicals which are
toxicologically similar. No definition of dissimilar action is presented in this EPA guidance.
11.4.
Agency for Toxic Substances and Disease Registry (2004)
In the “Guidance Manual for the Assessment of Joint Toxic Action of Chemical Mixtures” of
the Agency for Toxic Substances and Disease Registry (ATSDR 2004) simple similar action
is equated with dose addition “Dose Addition, also known as … simple similar action, …
assumes that the components of a mixture behave as concentrations or dilutions of one
another, differing only in their potencies. … Dose addition is considered most appropriate
for mixtures with components that affect the same endpoint by the same mechanism of
action.“ “Response Addition, also known as simple independent action … assumes that the
chemicals act independently and by different modes of action.”
11.5.
WHO/IPCS (2009)
The World Health Organisation/International Program on Chemical Safety presented in the
Harmonization Project Document 7: “Assessment of Combined Exposures to Multiple
Chemicals: Report of a WHO/IPCS International Workshop on Aggregate/Cumulative Risk
Assessment” (IPCS/WHO 2009) the following definitions: “Simple similar action refers to
chemicals that cause toxicity through a common toxic mode of action (MOA) and are thus
evaluated using dose addition approaches.“ “Simple dissimilar action is assumed when
chemicals cause a common health effect, but by a different toxic MOA. In this case, the toxic
responses are thought of as biologically and statistically independent events.“
11.6.
Norwegian Scientific Committee for Food Safety (2008)
The Norwegian Scientific Committee for Food Safety uses the following definition for
simple similar action in its report on „Combined toxic effects of multiple chemical
exposures” (VKM 2008): “simple similar action assumes that the compounds act on the same
biological site (e.g. receptor or target organ), by the same mechanism and that they differ
only in their potencies. Each chemical contributes to the toxicity of the mixture in proportion
to its dose, and their relative toxicities are assumed to be constant at all dose levels. The
Supporting publications 2012:EN-232
57
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
effect would be a result of the sum of the contributing dose of each chemical.” Exposure to
chemicals with simple similar action will result in dose addition. The committee defines
simple dissimilar action as follows “The chemicals contribute to a common result, but the
mechanisms by which the chemicals act are always different. Also, the nature and site of
action may possibly, but not necessarily, differ among the chemicals in the mixture.
Therefore, the presence of one chemical will not affect the toxicity of another chemical.”
Supporting publications 2012:EN-232
58
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 10: Comparison of the definitions given for ‘simple similar action’ and ‘simple
dissimilar action’
COT (2002)
EPA (2002)
ATSDR (2004)
EFSA (2008)
Simple similar action
chemicals act in the same way, by the
same mechanisms and differ only in their
potencies
Dose Additivity … chemicals that are
toxicologically similar and act at the
same target site
components affect the same endpoint by
the same mechanism of action and differ
only in their potencies
chemicals in a mixture act in the same
way, by the same mechanism/mode of
action and differ only in their potencies
Simple dissimilar action
modes of action and possibly the nature and
site of action differ
chemicals act independently and by
different modes of action
VKM (2008)
same biological site (e.g. receptor or
target organ), by the same mechanism
and differ only in their potencies
the modes of action and possibly, the
nature and sites of toxic effects differ
between the chemicals in a mixture,
chemicals
common result, but mechanisms are always
different; nature and site of action may
differ
WHO/IPCS (2009)
chemicals that cause toxicity through a
common toxic mode of action
chemicals cause a common health effect,
but by a different toxic mode of action
Supporting publications 2012:EN-232
59
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
12.
Legislation
12.1.
Consideration of possible cumulative risk in EU pesticide regulations
12.1.1. Legal background
EU legislation on pesticides has been substantially revised and harmonised in the recent
years. The evaluation, authorisation, placing on the market and control of plant protection
products in the EU are regulated by Council Directive 91/414/EEC (EU, 1991). This Council
Directive will be repealed by Regulation (EC) No 1107/2009 (EU, 2009) which will apply
from 14 June 2011.
These laws on the placing of plant protection products (PPP) on the market are complemented
by Regulation (EC) No 396/2005 (EU, 2005) on maximum residue levels (MRLs) of
pesticides in or on food, which aims at the harmonisation of MRL in the EU. Regulation (EC)
No 396/2005 covers the setting, monitoring and control of pesticide residues. There is a direct
interrelation between Regulation (EC) No 1107/2009 on the authorization of PPP and
Regulation (EC) No 396/2005 on pesticide residues: where appropriate the setting of MRL is
a prerequisite for granting authorization of a PPP (article 8, paragraph 1(g)). The same
interrelation exists with Council Directive 91/414/EEC (article 4, paragraph 1(f)).
In general, PPP have to be authorised before they can be put on the market. Only those PPP
can be authorised which contain an approved active substance, safener or synergist, i.e. which
are on the EU positive list. Before PPP are placed on the market it should be demonstrated
that they present a clear benefit for plant production and that they and their residues do not
have any harmful effect on human or animal health, including that of vulnerable groups, or
any unacceptable effects on the environment. Authorisation of PPP by an EU member state
should follow the uniform principles for evaluation and authorisation of PPP as presently
described in Annex VI of Council Directive 91/414/EEC, which still applies in the context of
Regulation (EC) No 1107/2009.
12.1.2. Consideration of cumulative risk in EU legislation
In the context of the approval procedure the risk assessment of the ingredients of PPP
predominantly addresses the risk arising from single substances. It is a basic requirement that
the information needed to establish, where relevant, an Acceptable Daily Intake (ADI),
Acceptable Operator Exposure Level (AOEL) and Acute Reference Dose (ARfD) is provided
by the manufacturer for an active ingredient, safener or synergist (see Annex II of Regulation
(EC) No 1107/2009).
Whereas Council Directive 91/414/EEC did not explicitly require cumulative risk assessment
this was introduced by the new Regulation (EC) No 1107/2009. Consideration of potential
mixture effects was a clear requirement for human or animal risk assessment of PPP and
residues. Plant protection products “shall have no immediate or delayed harmful effect on
human health, including that of vulnerable groups, or animal health, directly or through
drinking water (taking into account substances resulting from water treatment), food, feed or
Supporting publications 2012:EN-232
60
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
air, or consequences in the workplace or through other indirect effects, taking into account
known cumulative and synergistic effects where the scientific methods accepted by the
Authority to assess such effects are available; or on groundwater” (article 4, paragraph 3(b)).
The same applies to PPP residues (article 4, paragraph 2(a)): “they shall not have any harmful
effects on human health, including that of vulnerable groups, or animal health, taking into
account known cumulative and synergistic effects where the scientific methods accepted by
the Authority to assess such effects are available, or on groundwater“. For ecotoxicological
effects however, there is no requirement for taking into account known cumulative and
synergistic effects.
These more general terms in Regulation (EC) No 1107/2009 provide the basic legal
background for the consideration of potential cumulative risk of PPP ingredients without
addressing the necessary next steps. But Regulation (EC) No 396/2005 on maximum residue
levels clearly specifies in article 36, paragraph 1(c) the necessity to develop a methodology
for assessing aggregate, cumulative and synergistic effects (“studies and other measures
necessary for the preparation and development of legislation and of technical guidelines on
pesticide residues, aimed, in particular, at developing and using methods of assessing
aggregate, cumulative and synergistic effects”). Based on these requirements the Panel on
Plant Protection Products and their residues (PPR) developed and published a methodology
for the assessment of cumulative and synergistic risks of pesticides to human health (EFSA,
2008) which applies to substances with similar mode of action. It is the aim of this ongoing
project to develop a methodology for the cumulative risk assessment of substances with
dissimilar mode of action.
12.2.
Refusal of the authorisation of PPP
It is clearly stated in Regulation (EC) No 1107/2009 that active substances or products placed
on the market should “not adversely affect human or animal health or the environment”
(article 1, paragraph 4). As a basic requirement for the authorization all information necessary
for establishing guidance values such as ADI or AOEL have to be submitted to the authority.
In the context of the human health risk assessment for the general population or workers the
guidance values are compared with the estimated exposure towards the PPP or its residues
under the recommended conditions of use. If the exposure is lower than the ADI or AOEL,
the application of the PPP can be regarded as safe. However, if the authority concludes that
the application of the PPP as requested by the manufacturer is not safe for workers or the
general population the authorisation can take place but the authority can link it to certain
restrictions (article 6 of Regulation (EC) No 1107/2009). Those restrictions may comprise for
example:
•
manner and conditions of application,
•
designation of categories of users (professional or not professional),
•
designation of areas where the use may be authorised under specific conditions,
•
the need to impose risk mitigation measures and monitoring after use.
Supporting publications 2012:EN-232
61
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Which restrictions are appropriate has to be considered on a case by case basis and will also
depend on the importance of a PPP (risk benefit analysis). A similar procedure is applied for
environmental health risk assessment.
After authorisation of a PPP national authorities check in their monitoring programmes
whether the existing MRL are maintained. If these control programmes give any advice that
existing MRL cannot be maintained under normal conditions of use this might result in a
change of the authorisation and restrictions as discussed above or even the removal of the
authorisation may be the consequence (article 44 of Regulation (EC) No 1107/2009).
Neither Regulation (EC) No 1107/2009 nor Regulation (EC) No 396/2005 provides any
information about the legal consequences if a cumulative health risk assessment points to a
possible risk of the cumulative exposure. Even the EFSA opinion (EFSA, 2008) does not
provide any further information. Risk management measures to be taken in case of a
cumulative risk have not been worked out until now. It has to be discussed how to proceed if
a cumulative risk assessment (CRA) has indicated a potential cumulative risk.
•
Should all PPP, which were part of the CRA, be re-evaluated?
•
Should only those, which contribute most to the overall risk of the mixture, be reevaluated?
•
Should some PPP of the mixture be banned?
•
Should the manner of applications be changed?
•
Should the simultaneous use of certain PPP be banned?
Further guidance from the authorities on risk management measures in case of a risk due to
cumulative exposure is necessary.
Supporting publications 2012:EN-232
62
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.
Assessment concepts for mixture toxicity and empirical evidence for their
validity
Methods for experimental mixture studies can be divided into two major classes, “whole
mixture approaches” and “component based” approaches. In whole mixture approaches,
direct toxicological assessments of a given chemical mixture, such as a complex
environmental sample are conducted by treating a mixture as if it were a single chemical. The
composition of the mixture is not the topic of investigation, and whole mixture approaches do
not require new, mixture-specific assessment concepts.
In component-based approaches, efforts are made to anticipate the effects of a mixture on the
basis of the toxicity of its components. This makes it possible to draw more general
conclusions about the relationship between the effects of single substances and those of their
combinations. Numerous methods for this purpose have been described in the literature (see
the overview in (EC 2009). Such methods allow quantitative predictions of mixture toxicities,
without the need to test different mixture ratios, mixture concentrations or overwhelmingly
large numbers of permutations of mixture components. They require information about the
effects of the mixture components after administration as single chemicals and about their
levels in the mixture (mixture ratio). The effects of all components must have been measured
under the same conditions as the experimental mixture study, using the same toxicological
endpoint. The experimentally observed mixture effects can then be compared with those
expected on the basis of the effects of the components.
Among the component-based approaches, two fundamentally different concepts exist for the
calculation of mixture effects on the basis of the toxicity of its components, independent
action (IA) and dose or concentration addition (DA or CA). Both concepts rely on an
additivity assumption, which is based on the expectation that all chemicals in the mixture
exert their effects without influencing each other’s action. The additivity assumptions are not
fulfilled when components of the mixture interact with one another, e.g. by undergoing
chemical reactions with each other, or by inducing (de)toxifying metabolic conversions that
target some or all of the mixture components.
The difference between IA and DA is in the way in which each concept constructs its
additivity assumption. IA derives additivity assumptions from probabilistic considerations of
the effects of the mixture components. In contrast, DA is based on the idea that all
components in the mixture behave as if they are simple dilutions of one another, which is
often taken to mean that DA describes the joint action of compounds with an identical
mechanism of action.
Supporting publications 2012:EN-232
63
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.1.
Independent action (IA)
Independent action (sometimes also termed Effect Addition, Effect Multiplication or Abbotts
Rule) conceptualises mixture effects by assuming that combined effects can be calculated
from the effects caused by the individual mixture components on the basis of the statistical
concept of independent random events (Bliss 1939). This can be mathematically expressed as:
n
E ( cMix ) = 1 − ∏ [1 − E ( ci )]
i =1
(Eq. 1a)
if the effect increases with increasing concentrations (e.g. when mortality data are considered)
and
n
E (cmix ) = ∏ E (ci )
i =1
(Eq. 1b)
when the effect decreases with increasing concentrations (when e.g. survival rates are
observed). In both equations E(cMix) denotes the effect provoked by the total mixture at a
concentration
n
cMix = ∑ ci
i =1
.
E(ci) are the effects that the individual components would cause if applied singly at that
concentration at which they are present in the mixture.
Due to this probabilistic background, IA assumes strictly monotonic concentration-response
curves of the individual mixture components and an Euclidian-type effect parameter scaled to
an effect range of 0-1 (0-100%).
13.1.1. Applicability of IA to mixtures composed of agents with dissimilar modes of
action
Theoretically, the stochastic principles of IA are also valid when one and the same agent is
administered sequentially and irreversible events such as mortality are investigated. Because
organisms cannot die twice, the probability expressed in equation 1 a,b applies, despite the
fact that the mechanism by which the chemical provokes mortality is identical. In the case of
simultaneous administration of many chemicals the principle of independent events can only
be realised by making the additional assumption that all components in the mixture exert their
effects by activating different effector chains that converge to produce a common effect. This
is commonly thought to apply in cases where the chemicals in the mixture exert their effects
through strictly independent, i.e. dissimilar mechanisms. By activating differing effector
chains every component of a mixture of dissimilarly acting chemicals provokes effects
independent of all other agents that might also be present, and this feature lends itself to
statistical concepts of independent events.
Supporting publications 2012:EN-232
64
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.1.2. Data requirements for using IA
IA uses single substance effects, E(ci), for predicting a mixture effect (equation 1 a, b). This
means that the information need for utilising IA changes substantially with increasing
numbers of mixture components. For example, according to IA a binary mixture of two agents
that individually produce a 30% effect will lead to a 50% mixture effect. Thus, the application
of IA means that effects of 30% have to be measured with reliability which usually does not
present problems. In a 10-component mixture producing a 50% mixture effect, however, each
component has to be present at a concentration that produces only a 6.7% individual effect.
But effects of that magnitude are already at the borderline of what can be measured reliably in
many in vivo toxicological experiments. The more compounds are present in a mixture, the
lower the individual E(ci)’s become that are required as input values for estimating a 50%
mixture effect. The fact that increasingly lower E(ci)-values for each component need to be
measured for calculating IA-predictions is a serious drawback, as this increases experimental
demands beyond what is technically achievable with the number of animals per does group
normally used in toxicity studies.
NOAELs are not readily suited as input data for IA. NOAELs denote the highest tested doses
that produced effects not statistically significantly different from those in untreated controls,
but they do not describe effect magnitudes. Depending on the resolving power of the chosen
experimental arrangement, the effects associated with NOAELs can be quite large, but cannot
be measured directly, and are only accessible through regression modelling in dose-response
analyses. However, the number of doses tested in studies that establish a NOAEL is often
rather limited and does not permit regression analysis. As a result, it is normally not possible
to establish whether a NOAEL is associated with a 5%, 10% or 20% effect. Consequently, the
input data required for using IA are not accessible through reporting a NOAEL.
13.1.3. Under IA, when is a mixture risk acceptable?
Equations 1 a and b imply that agents present at doses associated with zero effects will not
contribute to the joint effect of the mixture. If this condition is fulfilled for all components in
the mixture no combination effect is expected under IA.
This means that claims of absence of mixture responses can only be substantiated if very
small effects can be distinguished with reliability from zero effects. However, especially with
mixtures composed of large numbers of components, this demands exceeds the resolving
power of most toxicological studies which struggle to resolve 5% effects. According to IA, 10
components at doses associated with 5% effect will already produce a combination effect of
45%. Correspondingly, 100 agents with a 1% effect are expected to produce a mixture effect
of 63%, and with 100 chemicals of 0.1% effect the expected joint response under IA will still
be 9.5 %. Such small effects can only be demonstrated with astronomically large numbers of
animals.
Supporting publications 2012:EN-232
65
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.1.4. Correlation assumptions in IA and their consequences
Consideration of the correlation of susceptibility of organisms in the IA model was often
present in early discussions, but has become less mentioned. Correlation of susceptibility
refers to the extent to which the individuals most susceptible to one chemical are also those
most susceptible to a second chemical. Correlation can be expressed as a correlation
coefficient, r, which takes values between 1 and -1. A value of 1 indicates that there is
complete correlation in susceptibility, i.e. that the sensitivity of individuals to either
component is the same, for example the individual most sensitive to the first chemical is also
the most sensitive to the second; conversely, a value of -1 indicates complete negative
correlation, in that the individuals most sensitive to one chemical will be the least sensitive to
the second chemical and vice versa. Finally, a value of 0 indicates that the susceptibility to
one chemical is unrelated to susceptibility to the other.
A generalised model of correlated independent action was introduced in the 1940s by Plackett
and Hewlett (reviewed in (Boedeker and Backhaus 2010)). Three subtypes of IA were
formulated for binary combinations. Firstly, when the susceptibility is uncorrelated, i.e. the
correlation coefficient (r) =0, the combined effect is given by the following equation, which is
equivalent to the IA equation presented in equation 1a (section 13.1) for a binary mixture:
When r=0,
P1,2 = P1 + P2 – P1P2
(Eq. 2a)
In this case, the mixture probability (P1,2) is given by adding the probability of an effect by
one chemical (P1) to the probability of an effect of a second chemical (P2) and subtracting the
product of the two probabilities (P1P2).
Secondly, when there is total positive correlation of susceptibility (r=1), the effect of the
combination is that of the most potent component because the individuals that might have
been affected by the weaker component will ‘already’ have been affected by the most potent
component. This situation is sometimes termed “no addition” (Boedeker and Backhaus 2010).
In this case the IA equation is:
When r=1,
P1,2 = max(P1, P2)
(Eq. 2b)
Thirdly, when there is total negative correlation (r=-1), the effect of the combination is the
numerical summation of both components, which is sometimes termed “effect summation”
(Boedeker and Backhaus 2010). In this case the individuals affected by either chemical are
different and so there is no need to consider effects of the two chemicals on one individual,
only on separate individuals:
When r=-1,
P1,2 = P1+P2
(Eq. 2c)
These formulations for binary mixtures depend on assumptions or knowledge about the
correlation between the two components in the binary mixture. Such knowledge is less likely
to be available, and assumptions harder to substantiate, for multi component mixtures with
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
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issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
tens or hundreds of components. For instance the concept of total negative correlation when
applied to a mixture of more than two components may not even be meaningful (EPA 2000).
The original formulation of these concepts, over 60 years ago, may underlie some suggested
pragmatic applications of IA. For example, the notion that the toxicity of a mixture is that of
the toxicity of the most potent component matches the formulation of IA under the
assumption of total positive correlation (Eq. 2b), whilst the notion that the mixture toxicity is
given by the summed effects of the components (effect summation, ES) matches the
formulation of IA under total negative correlation (Eq. 2c). Boedeker and Backhaus
considered that the subtype of IA with total negative correlation was “one of the rare
theoretical foundations “ of ES (Boedeker and Backhaus 2010), although total negative
correlation is not always stated as a requirement for the use of ES. The chief requirement
usually given for using ES is the presence of linear dose-response relationships (EC 2009).
The mathematical form of ES may also provide an approximation of the effect under IA when
certain other assumptions hold, for example that there are only two components and that their
individual effects are small (EPA 2000). In this case however ES is used as an approximation
to the effect under IA for ease of computation, not because it is considered innately valid.
In typical human toxicology situations, it seems unlikely that there will be sufficient
information about the correlation of susceptibilities to the mixture components. In the absence
of this information, the assumption of no correlation may be more reasonable that assuming
the extremes of either total negative or total positive correlation. In the event that assumptions
of correlation are made, this should be clearly stated and the rationale for doing so should be
justified.
13.1.5. Empirical evidence for IA
The results of the systematic literature search were carefully examined for empirical evidence
for the validity of IA. In particular we sought examples when:
•
IA provided an accurate prediction of a mixture effect, in a situation in which the
predictions under DA and IA were separable.
•
IA provided a prediction that was more conservative than the DA prediction.
•
IA provided a more conservative prediction AND was also accurate.
We found that experimental studies published since the State Of The Art Report on Mixture
Toxicity (EC 2009) have not altered the literature summary given in that report. The current
position is now summarised.
The ecotoxicology literature contains a few examples when the effects of carefully designed
mixtures were shown to validate the IA model (EC 2009). Of importance is a study of 16
biocides whose combination effect on algal toxicity was accurately predicted by IA (Faust et
al. 2003). These components were selected on the basis of their strictly different specific
mechanism of action, which are listed in the CRADIS database analysis of this study. Faust et
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Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
al. also found that whilst IA was the accurate model, in this case the predicted effect under
DA was greater than that under IA, and so DA could be considered the conservative model. If
this finding is generally applicable then there would be reassurance that even in cases when
IA is valid, DA can still provide a conservative risk estimate, supporting the use of DA as the
default concept in mixture risk assessment.
A proof-of-principle example of the validity of IA has not been identified in the mammalian
toxicology literature (this report, (EC 2009)). The reason for this is likely to be the
difficulties, including costs and ethical considerations, of performing mammalian studies with
a sufficiently large number of components (which is required to allow the predictions of IA
and DA to be distinguished from each other) and the difficulty in selecting an appropriate
effect for which there are enough well characterised chemicals with strictly different specific
mechanisms of action. The amount and level of knowledge required to design a mixture
experiment that is suitable to test the hypothesis that IA is accurate in a mammalian system
appears to be far greater than the knowledge that is typically available for chemicals.
A situation when IA was both more conservative than DA, and also accurate was not
identified in the literature. The factors that lead to IA producing a more conservative
prediction than DA are discussed in section 13.4.This putative situation is an important one to
consider, since it would have implications for the use of DA as a conservative default.
Consequently the factors discussed in section 13.4 can be evaluated to consider how likely it
is that this situation could occur, given that it has not been observed in experimental studies to
date.
13.2.
Dose addition (DA)
DA (also known as concentration addition, CA) is based on the idea that all components in
the mixture behave as if they are simple dilutions of one another, which is often taken to
mean that DA describes the joint action of compounds with an identical mechanism of action.
When chemicals interact with an identical, well-defined molecular target, it is thought that
one chemical can be replaced totally or in part by an equal fraction of an equi-effective
concentration (e.g. an EC50) of another, without changing the overall combined effect. If the
assumption of dose addition holds true, these fractions of equi-effective single substances
concentrations – also called toxic units – simply sum up to an overall toxic unit of the
mixture. Therefore, DA is also known as “Toxic Unit Summation”. The concept can be
mathematically formulated as:
ECxMix
⎛ n p ⎞
= ⎜∑ i ⎟
⎝ i =1 ECxi ⎠
−1
(Eq. 3)
with n denoting the number of mixture components, pi the relative fraction of chemical i in
the mixture, and x a common effect level, provoked by an exposure to a single substance or
mixture concentration ECxMix resp. ECxi.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
In general, no explicit formulation of the DA-expected mixture effect E(cMix) is possible.
Direct calculations are restricted to the effect levels associated with the effect concentrations
(ECx-values) (Faust et al. 2003).
The requirement of parallel dose response curves has often been used as a decision criterion
about the application of DA to a specific mixture. However, the general formulation of DA in
equation 3 does neither assume a specific shape of each concentration-response curve of the
components, nor a specific relationship between the curves, such as parallelism. Even if all
chemicals in a mixture share an identical receptor binding site, differences e.g. in the
toxicokinetic behaviour of the substances might lead to concentration-response curves that are
not parallel, yet DA may still apply.
13.2.1. Data requirements for using DA
To predict mixture effects by using DA, information about the doses that induce the same
specific effect are required for both the mixture and all single components. For example, if the
effect dose of a mixture leading to a 50% effect is known, then the equivalent effect doses
(ED50) for all mixture components need to be available to reach decisions whether the
combined effect is dose additive (see equation 3). In this case, the sum of toxic units in
equation 3 will be 1. The same requirements need to be met for any other effect level.
Usually, information about effect doses is accessible through dose-response analyses of the
individual components in a mixture.
As with IA, NOAELs are strictly speaking not suited as input values for using DA, because
NOAELs represent different (but unknown) effect doses. However, unlike IA, the
measurement precision required for using the concept as the number of mixture components
increases, does not change. This feature makes DA generally easier to use in most situations.
It is obvious from equation 3 that DA represents the weighted harmonic mean of the
individual ECx values, with the weights being the fractions pi of the components in the
mixture. This has important favourable consequences for the statistical uncertainty of the DApredicted joint toxicity. As the statistical uncertainty of the DA-predicted ECx for the mixture
is the result of averaging the uncertainties of the single substance ECx-values, the stochastic
uncertainty of the DA prediction is always smaller than the highest uncertainty found in all
individual ECx-values. Perhaps contrary to intuition, the consideration of mixtures composed
of a large number of agents actually reduces the overall stochastic uncertainty. This feature
renders DA predictions quite reliable and robust.
13.2.2. Under DA, when is a mixture risk acceptable?
DA implies that every toxicant in the mixtures contributes in proportion to its toxic unit (i.e.
its concentration and individual potency) to the mixture toxicity. Whether the individual
doses are also effective on their own does not matter. Thus, combination effects should result
from toxicants at or below effect thresholds, provided sufficiently large numbers of
components sum up to a sufficiently high total dose. Unfortunately, that is often
misunderstood to mean that mixture effects will arise with any combination of agents, if the
principles of DA are fulfilled. However, this is not the case. For example, the joint effect of
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
two agents combined at 1/10 of their ADI is expected to be considerably smaller than the
effect (if any) associated with the ADI of each of the chemicals on their own. Similarly, 100
chemicals combined at 1/100 of their ADI will not produce a mixture effect greater than the
effects provoked at the ADI’s of each of the single components (see equation 3).
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.3.
The role of mode and mechanism of action information in the choice between IA
and DA
We searched and analysed several sources of guidance on mixture risk assessment for insights
into criteria relevant to the choice between similar and dissimilar action, dose addition or
independent action. An overview of the guidance from regulatory bodies is provided in Task
4 (section 14), and this section concentrates on the potential role of mode of action in mixture
risk assessment.
13.3.1. Definitions of mode and mechanism of action
Two concepts that can feature in guidance on the application of mixture concepts are mode of
action and mechanism of action. These concepts have been defined as follows:
Mode of action (MOA): a “biologically plausible sequence of key events leading to an
observed effect supported by robust experimental observations and mechanistic data” (Boobis
et al. 2006).
Mechanism of action: “a sufficient understanding of the molecular basis for an effect and its
detailed description so causation can be established in molecular terms” (Boobis et al. 2006)
or “a detailed explanation of the individual biochemical and physiological events leading to a
toxic effect” (EFSA 2008b). It has been noted that the US EPA use the term “mechanism of
action” to accompany the definition of “mode of action” given above (EFSA, 2008b). Care
must therefore be taken not to confuse these concepts when comparing international,
European and American activities.
As an example of an MOA definition, the key events making up the MOA of a DNA-reactive
chemical are:
“1. Exposure of target cells (e.g., stem cells) to ultimate DNA-reactive and mutagenic
species—in some cases this requires metabolism.
2. Reaction with DNA in target cells to produce DNA damage.
3. Misreplication on damaged DNA template or misrepair of DNA damage.
4. Mutations in critical genes in replicating target cell.
5. These mutations result in initiation of new DNA/cell replication.
6. New cell replication leads to clonal expansion of mutant cells.
7. DNA replication can lead to further mutations in critical genes.
8. Imbalanced and uncontrolled clonal growth of mutant cells may lead to preneoplastic
lesions.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
9. Progression of preneoplastic cells results in emergence of overt neoplasms, solid tumours
(which require neoangiogenesis), or leukaemia.
10. Additional mutations in critical genes as a result of uncontrolled cell division results in
malignant behaviour” (Preston and Williams 2005).
This example shows that even an MOA, compared to a full mechanism, can contain a
significant amount of detail, and illustrates the data demands that would be required to
systematically analyse multiple chemicals with multiple modes of action. A fully elucidated
mechanism of toxicity is only available for a few chemicals, one example being cyanide,
whilst more chemicals have a known MOA, with key events that are “known, measurable,
necessary and consistent” (Carmichael et al. 2011). The studies required to identify an MOA
are not currently a regulatory requirement for industrial chemicals, perhaps because such
studies have a uncertain time scale and costs, with inconsistent interpretation and regulatory
impact (Carmichael et al. 2011).
Our analysis (sections 10.1 and 10.3.1) showed that when authors of experimental studies
have made reference to mode or mechanism of action, they are not using a common
framework to make these assignments. Approaches using agreed MOAs, or at least MOAs
defined in accepted ways, could provide a common framework that, if used, would improve
comparison of results from mixture assessments and experimental studies. A number of
frameworks are being proposed and may prove useful if they achieve consensus. However,
their suitability for use in assigning mixtures to DA or IA must also be assessed.
IPCS frameworks for assessing the relevance of cancer (Boobis et al. 2006) and noncancer
(Boobis et al. 2008) modes of action have been developed. The frameworks apply a weight of
evidence approach based on the Bradford Hill criteria for causality to evaluate a proposed
MOA and its relevance to humans. Such frameworks provide a mechanism to establish
whether the MOA of a chemical has been described previously for other chemicals or whether
it is novel. Of relevance to CRA, the systematic use of such frameworks, and MOAs arising
from or validated by them, might allow the development of a set of MOAs founded on the
same principles, thus allowing them to be compared, and that can be used for more reliable
grouping or related activities. It was suggested that a database of generally accepted MOAs
and informative cases should be constructed and maintained (Boobis et al. 2006). This need
has been recognised by others, for example McCarty and Borgert considered that “the
absence of any generally accepted classification scheme for either modes/mechanisms of
toxic action or of mechanisms of toxicity interactions is problematic as it produces a cycle in
which research and policy are interdependent and mutually limiting.” (McCarty and Borgert
2006). Clearly, this issue constitutes a substantial knowledge gap.
The advantage of these approaches is that their international nature may favour international
acceptance and agreement, however are they suitable for the purpose of assigning DA or IA?
We note that the definitions of mode or mechanism of action given above, do not define when
a mode/mechanism is novel, or separate or distinct from another mode/mechanism. This is a
critical need if the mode/mechanism information is to be used to inform the selection of a
mixture assessment concept, DA or IA. If the modes/mechanisms are not independent the
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
case for IA will not be supportable, and if no reference to independence is made then the
definition will not be applicable to the choice between concepts, since independence is the
key criteria for choosing IA over DA.
13.3.2. Use of empirical evidence to infer similarity or dissimilarity
Some authors have concluded that, rather than selecting a mixture concept, DA or IA, and
using it to predict the effect of a mixture, in fact one can perform a mixture experiment and
then infer knowledge about the similarity or dissimilarity of the components of a mixture
based on whether its results are closer to the DA prediction (infer that components are
similar) or to the IA prediction (infer that the components are dissimilar).Example of this
usage are now provided:
Walter et al. stated "These findings [that experimental effect was predicted by IA] strongly
indicate, that the modes of action of mixture components are indeed dissimilar in a way, that
is sufficient to predict the mixture toxicity with the concept of independent action." (Walter et
al. 2002).
Bellas concluded that the toxicity of a mixtures of zinc pyrithione and Sea-Nine “...was
accurately predicted by the IA concept, suggesting a dissimilar mode of action of those
substances” (Bellas 2008)
Hodges et al. titled their paper” Defining the toxic mode of action of ester sulphonates using
the joint toxicity of mixtures.” and concluded that “...data indicated that ES substances exhibit
concentration addition with linear alkylbenzene sulphonate (LAS) and phenols and response
addition with alcohols. This suggests that ES behave with a similar mode of action to phenol
and LAS which are known polar narcotics and with a dissimilar mode of action to alcohols
which are known baseline narcotics.”(Hodges et al. 2006).
Escher et al. proposed an in vitro assessment of modes of toxic action, relating to
pharmaceutics in aquatic environments and used mixture experiments “as a diagnostic tool to
analyse the mode of toxic action” (Escher et al. 2005).
Lutz et al claimed that “Distinction between dose addition and response addition for the
analysis of the toxicity of mixtures may allow differentiation of the components regarding
similar versus independent mode of action” (Lutz et al. 2005).
This approach reverses the use of mixture concepts to predict effects, and may therefore be
less relevant to the needs of CRA. However, it illustrates one possible response to the
difficulties in assigning similarity and dissimilarity, namely that the assignment is not done.
This post hoc assignment of similarity or dissimilarity would have an impact in the methods
used to assess the relative impact of individual components in a mixture to identify targets for
risk management. On consequence to CRA, is that this approach would require experimental
data for the mixture of concern.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.3.3. Conclusions
Existing guidance can depend heavily on information, e.g. MOA, which is often not available
or is available in incompatible forms. Frameworks are being developed, but international
consensus is required. A clear requirement for the framework to be suitable for the choice
between DA and IA would be advantageous.
Criteria based on concepts such as MOA were not framed in order to be useful for guiding the
choice between mixture concepts, such as DA and AI; consequently they may indeed have
limited usefulness for such purposes. Known or plausible independence is the key criteria for
using IA instead of DA. We also note that, for the IA model, non-similarity is not the same as
dissimilarity. This is reflected in some guidance (e.g. (EPA 2000)) by the use of similarity vs.
independence, rather than similarity vs. dissimilarity. This is a helpful distinction.
The clear difficulties with using the available frameworks and definitions (lack of consensus,
lack of suitability for choosing (independence), high data requirements and poor data
availability for the ‘world’ of chemicals) lead to the questions: how important is it to be able
to choose between DA and IA, given that it is not easy to do so? And what is the impact of
having to assume one or other concept? These questions are addressed in detail in the
following section (section 13.4).
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.4.
Quantitative differences between DA and IA predictions
13.4.1. Problem formulation
In general, the assumption of dose additivity (DA) is considered to provide a more
conservative estimate of mixture toxicity than the alternative assumption of independent
action (IA). Synergistic effects that exceed the DA expectation are exemptions and not the
rule, at least for multi-component mixtures. In addition, data requirements for a proper
application of DA are easier to fulfil than for IA. With these three arguments, DA has been
recommended as a reasonable worst case assumption for the purpose of regulatory hazard and
risk assessments (EC 2009), in particular in situations, where modes and mechanisms of
action of mixture components are not fully independent and not fully dissimilar, or
insufficiently known or unknown. Similarly, the recently published WHO/IPCS framework
(Meek et al. 2011) suggests DA as a default tier zero assumption for all components cooccurring in an exposure scenario and potentially contributing to a common adverse health
outcome.
The main concern that may be raised against such a use of DA as a pragmatic default
assumption, irrespective of toxicants modes and mechanisms of action, is that the use of a
conceptually unsound model may potentially result in vastly over-protective mixture toxicity
assessments, not scientifically justified and conflicting with the principle of proportionality in
the regulatory management of chemicals risks.
This raises the question about the quantitative differences between independent action and
dose addition:
•
What is the maximal quantitative error that may result, if DA is applied in a situation
where in fact IA or a mixed model would provide the correct mixture toxicity
estimate?
To explore this question further, it is necessary to define a suitable measure for the
differences in mixture toxicity predictions by DA and IA. In general, toxicity may be
quantified either in terms of the strength or frequency of an effect (E) at a given dose (d) or
concentration (c), or in terms of the dose or concentration causing a specific effect level (x),
so-called effect doses (EDx) or concentrations (ECx), such as ED50 for instance. In a
regulatory context, usually the latter approach is used. Correspondingly, effect doses (or
concentrations) can be calculated for mixtures that contain a given set of toxicants in a given
dose ratio (EDxmix). To distinguish between the models used for their calculation, either DA
or IA, they are in the following denoted by EDxDA and EDxIA, respectively. Such effect doses
may potentially vary over orders of magnitude. It is therefore more convenient to describe the
differences between EDxDA and EDxIA in terms of a relative figure rather than in absolute
terms. Considering further that EDxDA is usually expected to be a lower value (i.e. the higher
mixture toxicity estimate) than EDxIA, the differences between both predictions are in the
following quantified my means of the ratio:
•
EDxIA / EDxDA
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
This quotient is 1 in the case that both concepts give identical predictions, it is >1 in the
typical case that IA predicts a lower toxicity (i.e. a higher effect dose), and conversely is
between 0 and 1 if IA predicts higher toxicity. In other words:
•
The ratio EDxIA / EDxDA gives the factor by which dose addition overestimates the
actual toxicity in a situation where independent action would in fact provide the
correct estimate.
13.4.2. Empirical evidence
In published experimental studies on multi-component mixture toxicity the quantitative
differences between both predictions, IA and DA, have been reported to be remarkably small,
at least from a regulatory perspective. For different types of mixtures with up to 20
components, predictions of EC50 or ED50 values derived from the two models differed in no
case by more than a factor of 5 (EC 2009). From these pieces of evidence the quantitative
differences between IA and DA may be assessed to be of only minor relevance in a regulatory
context. However, it may be questioned, whether the available observations do really
represent typical situations for realistic exposure and assessment situations. Their validity
may be restricted to the special mixtures, conditions and toxicity endpoints which have
actually been investigated. Therefore, a consensual acceptance of DA as a generally
justifiable default assumption would require strong arguments against this suspicion. Hence,
in the light of available experimental evidence, the question to be clarified can also be
formulated as follows:
•
Do existing experimental findings of relatively small differences between DA and IA
just represent special situations or do they reflect a general rule?
This problem cannot be solved by experimentation only but needs complementary
approaches, such as mathematical and statistical analyses and simulation studies. The basic
questions to be addressed by such approaches are:
•
Which are the factors that determine the quantitative differences between independent
action and dose addition?
•
May the ratio EDxIA / EDxDA take any value or do limiting cases exist?
•
Which ratios between both predictions may occur under realistic scenarios?
13.4.3. Limiting factors and resulting maximal differences
For calculating predictions of effect doses or concentrations of mixtures, the general
mathematical definitions of DA and IA can be transformed into the following expressions, as
explained in detail in Faust et al. (2003).
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
DA:
EDxDA
⎛ n
p ⎞
= ⎜ ∑ −1 i ⎟
⎝ i =1 Fi ( xi ) ⎠
−1
(Eq. 4)
n
x % = 1 − ∏ (1 − Fi ( pi • ( EDxIA )) )
IA:
(Eq. 5)
i =1
With these two formulae, the parameters that determine the ratio between predictions of effect
doses by IA and DA are completely defined. These are simply the variables for which input
data have to be entered into the formulae for obtaining the alternative mixture toxicity
predictions EDxDA and EDxIA. There are four crucial factors:
•
the number of mixture components n,
•
the slope of the individual dose response curves defined by functions Fi ,
•
the mixture ratio p1 : p2 : ... : pi , with pi denoting the individual dose di of component
i expressed as percentage of the total dose Σdi of a mixture (pi = di/Σdi; Σpi = 1; i = 1
to n), and
•
the effect level X under consideration.
If no restrictions apply to these parameters, there is no fixed type of relation between both
predictions of mixture toxicity. The effect dose calculated under the assumption of
independent action (EDxIA) may be larger, equal to, or even lower than the corresponding
prediction by DA (EDxDA) (Drescher and Bodeker 1995). Practical relevance has been
demonstrated for the first two situations (EC 2009) but appears to be questionable for the lastmentioned theoretical case. To our knowledge, there is no convincing experimental example,
where IA does not only predict a significantly higher toxicity than DA, but where this
prediction is also the more accurate one.
Although the ratio between both predictions is fully determined by the four parameters, it is
unfortunately not possible to express the ratio EDxIA / EDxDA as an explicit function of these
parameters. This results from the fact, that the formulation of IA as given in Eq. 5 provides
only an implicit prediction of the effect dose of a mixture, which cannot be turned into an
explicit function. Hence, the same applies to the ratio EDxIA / EDxDA.
Nevertheless, it is possible to examine the ratio EDxIA / EDxDA for the existence of limit
values that cannot be exceeded under a certain constellation of determining factors. To this
end, the only general presumption to be made is that the dose response functions Fi of
individual toxicants are monotonously increasing, and that any given effect Ei can be
definitely related to a single dose, the so-called effect dose EDx, which can only take positive
values (EDx ≥ 0). If dose response functions include lower or upper thresholds below or
above which the effect is constantly zero or 100 %, these thresholds must therefore be defined
as ED0 and ED100, respectively.
Supporting publications 2012:EN-232
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.4.3.1.The number of mixture components (n)
Given the aforementioned premises, limiting cases exist for independent action: the predicted
effect dose of the mixture can never be larger than the sum of individual effect doses of
mixture components that cause the same effect level X:
n
0 ≤ EDxIA ≤ ∑ EDxi
(Eq. 6)
i =1
As a consequence, limiting cases also exist for the ratio between both predictions,
EDxIA / EDxDA:
n
∑ EDx
i
EDxIA
i =1
0≤
≤
−1
EDxDA ⎛ n
pi ⎞
⎜ ∑ −1
⎟
⎝ i =1 Fi ( xi ) ⎠
(Eq. 7)
As was shown in (Faust 1999), this expression can be solved to
0≤
EDxIA
≤n
EDxDA
(Eq. 8)
This means, for a mixture with a given number of components n, the ratio EDxIA / EDxDA
cannot take any value, but is generally delimited by zero and n, irrespective of all other
parameter values. In other words:
•
Effect doses predicted by independent action can never exceed the corresponding
prediction by dose addition by a factor that is greater than n, i.e. the number of
mixture components.
This means, at least theoretically, that the ratio can become infinitely small. But for a given
number of components it cannot become infinitely large. For a two-compound mixture a
maximal factor of 2 applies, and for mixtures with up to ten components it can never exceed
an order of magnitude. For multi-component mixtures with very large numbers of
components, however, it may still become very large, at least theoretically if no restrictions
apply to the other determining factors.
13.4.3.2.The dose ratio of mixture components
Further analysis reveals that the maximum ratio of n can only occur under the condition that
all components of a mixture are present in equal fractions of equi-effective doses. In any other
situation the ratio will always be smaller than n.
When doses or concentrations of mixture components are expressed as fractions of equieffective doses or concentrations, the resulting dimensionless dose units have been termed
toxic units (TU) (Sprague 1970). For practical applications, mostly the 50% effect level is
used as the reference point (ED50i). In principle, however, TU values can be calculated for
any effect level X:
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TUxi =
di
EDxi
(Eq. 9)
Given these definitions, it can be shown that the maximum ratio between the predictions
based on IA and DA is defined by the ratio between the sum of toxic units and the maximal
toxic unit value of the single substances in a mixture (Junghans et al. 2006):
n
TU i
∑
EDxIA
i =1
0≤
≤
≤n
EDxDA
max {TU i }
(Eq. 10)
i∈(1,..., n )
This has important practical implications for situations where one or a few compounds
dominate a mixture in terms of toxic units. For example, if one mixture component
contributes already 50% to the total sum of toxic units, the quotient ECxIA / ECxDA can never
exceed a value of 2, no matter what the total number of components and their toxic units may
be.
13.4.3.3.The slopes of dose response curves
Independent of the limiting effect of a specific dose ratio, the slopes of individual dose
response curves of a given set of toxicants in a mixture have a general limiting effect on the
possibility range of the prediction ratio EDxIA / EDxDA. The relationship between slope and
prediction differences is visualized in Figure 6 for a hypothetical 10-compound mixture. The
ratio EDxIA / EDxDA takes the value of 1, i.e. both IA and DA give exactly the same prediction
(Figure 6, B), if the dose response curves of all mixture components can be described by the
following specific form of the Weibull function (Drescher and Bodeker 1995)
E (di ) = 1 − exp(− exp(α i + ln(10) • log(di ))) ,
(Eq. 11)
in which the general slope parameter βi has the special value of ln(10) (= 2.3025…), while αi
is a location parameter that has no effect on the ratio EDxIA / ECxDA. If all dose response
curves are steeper, DA always predicts a higher toxicity than IA (Figure 6, A) and the
maximum possible ratio EDxIA / EDxDA tends towards the maximum value of n (= number of
components) if all the curves become infinitely steep. Conversely, DA predicts a lower
mixture toxicity than IA (Figure 6, C), and the lowest possible ratio tends towards zero if the
individual dose response curves of all mixture components become infinitely flat, and if no
threshold assumptions are made (see section 14.6.3.4 below). In a mixed situation with both
relatively steep and flat curves (as compared to Eq. 11), large prediction differences are
unlikely to occur (see section 13.4.4 below) and it strongly depends on the dose ratio and the
effect level, whether DA or IA predicts the higher mixture toxicity (Figure 6, D).
The finding, that the steepness of individual dose response curves is a crucial limiting factor
for the possible quantitative differences between mixture toxicity predictions by IA and DA,
leads to the question, what are realistic scenarios for the distribution of slope values for dose
response curves? In experimental reality, dose response curves become neither infinitely steep
nor infinitely flat, but their slopes typically vary in a certain range, which unfortunately,
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
however, is neither very well understood nor documented. Therefore, this point needs further
research that goes beyond the tasks of this report. However, as an explorative step in this
direction, we performed an initial simulation study with a sample set of eco-toxicological
dose response data for algal reproduction (section 13.4.4 below).
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Individual Dose Response Curves
Mixture Toxicity Predictions
(ED50 Ratio)
A All βi = 10
100
DA
% Effect
IA
50
0
0.00001
% Effect
100
0.001
0.1
10
1000
0.00001
0.1
10
1000
B All βi = LN(10)
IA congruent with DA
50
0
0.00001
100
0.001
0.001
0.1
10
1000
0.00001
0.001
0.1
10
1000
0.1
10
1000
0.1
10
1000
C All βi = 1
DA
% Effect
IA
50
0
0.00001
0.001
0.1
10
1000
0.00001
0.001
D Different βi
100
DA
% Effect
IA
50
0
0.00001
0.001
0.1
10
Dose (Log Scale)
1000
0.00001
0.001
Total Dose (Log Scale)
Figure 6: Effect of the slope of dose response curves (DRCs) of individual substances
(left) on the ratio between predictions of mixture toxicity by IA and DA
(right). Dose response curves of 10 hypothetical mixture components are
described by the Weibull model Ei = Fi(di) = 1-exp(-exp(α+βi•log10(di)))).
Different scenarios are depicted for the slope parameter βi: steep curves (A),
intermediate steepness (B), very shallow curves (C), and a mixed situation (D).
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Resulting mixture toxicity predictions by DA and IA are shown for mixtures
containing the 10 components in the ratio of their individual ED50 values.
13.4.3.4.The effect of threshold assumptions
The situation, that a general upper limit (= n) can be defined for the ratio EDxIA / EDxDA, but
no general lower limit other than zero, results from the fact that a priori no restrictions were
assumed for the individual dose response curves other than that they are monotonously
increasing. The situation changes, when threshold assumptions are made.
If threshold doses or concentrations (ED0), at and below which the response is assumed to be
either zero or constant and indifferent from controls, are assumed to exist for individual
mixture components, than both models, IA and DA, imply that such thresholds should also
exist for mixtures. Under the assumption of IA, no mixture toxicity occurs, if doses of all
components are at or below individual threshold levels. Under the assumption of DA, in
contrast, this is only a necessary but no sufficient condition. DA means that mixture toxicity
will in no case occur, if the doses of all components are not higher than 1/n of the individual
threshold (with n being the number of components). If, in contrast, individual doses are in the
range between 1/n and 1/1 of an individual ED0, DA implies that it depends on the dose ratio
whether an effect occurs or not.
As a consequence of these different model assumptions, effect doses predicted by IA can
never be lower than a threshold for mixture toxicity calculated under the assumption of DA
(ED0,DA). Hence, threshold assumptions result in a lower limit for the generally possible range
of ratios EDxIA / EDxDA (Figure 7) which is then defined by
ED 0 DA EDxIA
≤
≤n
EDxDA EDxDA
(Eq. 12)
Hence, the theoretical possibilities for situations where IA predicts a considerably higher
mixture toxicity than DA are substantially narrowed down by the introduction of threshold
assumptions and largely confined to higher effect levels.
In addition to defining a lower boundary for the ratio the EDxIA / EDxDA, threshold
assumptions may also have a quantitative impact on the calculation of effect doses under the
assumption of IA. These predicted effect doses may become higher and consequently also the
difference between IA- and DA-based predictions may be increased. This is the consequence
of methodological constraints of statistical dose-response modelling as explained in the
following.
Application of the IA model for multi-component mixtures requires reliable statistical
estimates of low effects of individual mixture components. If a statistical methodology is used
which tends to over- (or under-) estimate low effects, then it is very likely that the overall
mixture effect that is expectable under the assumption of IA is in the same way over- (or
under-) estimated. In a strict quantitative sense, classical regression-based dose-response
models always estimate effect changes in comparison to a control or reference value,
independent of the dose range. They assume that with decreasing doses or concentrations,
effect changes are becoming microscopically small and thus negligible, but not zero (unless
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
the dose is also zero). This model assumption obviously conflicts with the threshold concept
in toxicology, which assumes that below a certain threshold dose no effect changes occur at
all. Continuous regression models still provide estimates of effect changes (although very
small) at dose ranges below the assumed threshold doses. As a result of this methodological
constraint, it is possible that a combination effect is overestimated by applying the IA model
with dose response estimates from continuous regression models, when at least one
compound in the mixture is present below its toxicological threshold.
Figure 7: The potential range of ratios between predictions of effect doses for mixtures
by IA and DA. A: Under the assumption of continuous dose response curves
(DRCs) for single substances. B: Under the assumption of thresholds for
individual DRCs.
The problem of quantifying toxicological thresholds is complex, both conceptually and
methodologically. If the term threshold is understood as a zero effect dose, its quantitative
determination from experimental data by statistical means is for various reasons impossible
(see (Scholze and Kortenkamp 2007;Slob 1999). So-called mathematical threshold models are
therefore sometimes used as a pragmatic approximation for the toxicological threshold (Cox
1987). They are derived from commonly used nonlinear regression models by additional
inclusion of a model parameter which describes the threshold dose. These threshold models
are fitted to the data by the same statistical procedures as non-threshold models and provide
an estimate of the most likely dose-response curve (Figure 8). The basic mathematical model
may be chosen from a large class of models and different methods are also available for the
estimation process. There is no model and method that is the universally most powerful and
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
most appropriate one. Consequently, the resulting threshold estimates depend crucially on the
selected model and method. Moreover, if not bound to positive ranges, the estimation process
can indeed also result in a negative threshold, simply because the model would then describe
the data best. It is also not well understood how the quality and quantity of data (such as dose
or concentration spacing, replicate number, and data variation) influences the resulting
threshold estimates; however, to some degree data uncertainty is reflected by the confidence
belts of the threshold estimates.
With the aim of developing ideas about the potential quantitative effect that the use of
threshold models may have on the ratio between alternative predictions of mixture toxicity by
IA and DA, we applied such models as part of the following simulation study with a sample
set of ecotoxicological dose response data. As can be seen in Figure 8, threshold models may
predict zero effect levels for doses that are associated with considerable effects in continuous
dose-response models.
Figure 8: Classical continuous dose-response model (black line) and threshold model
(red line), both fitted to the same set of data (grey dots)
13.4.4. Sample calculations and simulations
The mathematical considerations in the preceding sections demonstrate that, if no specific
dose ratio is fixed, the range of possible differences between IA and DA is limited by the
number of mixture components in the first place, and that it may be further restricted by the
actual slope of dose response curves of single substances in the second place. Thus, slope
turns out to be a crucial factor, but unfortunately, in the open literature, results of toxicity
testing are mostly reported in terms of effect doses or concentrations (e.g. ED50) and NOEL
or LOEL values only. Although the situation has somewhat improved in recent years,
typically, neither slope parameters nor original experimental data, allowing a re-fitting to
appropriate regression models, are documented.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The following sample calculations and simulations therefore make use of a high quality
sample set of ecotoxicological data for the inhibition of algal reproduction. These of course
have no direct relevance for human toxicology. However, according to the experience of the
authors with the statistical analysis of actual data from chronic mammalian toxicity studies
(Christiansen et al. 2008;Christiansen et al. 2009;Hass et al. 2007;Metzdorff et al. 2007), it
can be stated that the dose response curves resulting from such studies are typically not
steeper than those observed for algal reproduction. On the contrary, dose response curves for
reproductive toxicity in rodents often appear to be less steep. As explained in the preceding
sections, this would mean that quantitative differences between predictions of mixture
toxicity by IA and DA can only be expected to be typically smaller than those observed in
simulations with algal toxicity data, not larger. Thus, if relatively small differences between
mixture toxicity predictions by IA and DA are observed with the algal toxicity data, this gives
good reason to hypothesize, that the same might apply to estimates of chronic mixture toxicity
in rodents. In this sense, the sample simulations were intended to provide a proof-of-principle
type of evidence.
The data set comprised original experimental data for the algal toxicity of 106 different
chemicals, mainly pesticides, few anti-foulings, anti-biotics, and surfactants and some
industrial chemicals. The data were compiled and generated as part the EU FP5 Project
BEAM (Bridging effect assessment of mixtures to ecosystem situations and regulations,
EVK1-CT-1999-00012, 2000-2003), specifically with the aim of obtaining high quality
descriptions of dose response curves that allow statistically valid estimates of low effect
concentrations for the purpose of low dose mixture toxicity studies (Faust et al. 2001;Faust et
al. 2003;Walter et al. 2002). To this end, they were all generated under identical testing
conditions and with a considerably higher number of test concentrations than in usual
regulatory testing protocols. Summary statistics derived from these data sets have been
published in variety of papers; for the purpose of the simulations performed for this report,
however, exclusive use was made of the original experimental raw data, which were all refitted to a variety of both continuous and threshold response models.
To study the potential range of ratios between predictions of effect concentrations for the
algal toxicity of mixtures that can be generated from 106 different chemicals, computer
simulations were performed in two steps: first a deterministic simulation study on the possible
extreme values for the prediction differences was conducted, and secondly a probabilistic
simulation study on the distribution of ratios between predictions based on IA and DA was
performed, which also includes a comparison of the use of continuous dose response models
with threshold models.
13.4.4.1.Deterministic simulations of extreme prediction differences
Deterministic simulations were performed to calculate the extreme ratios between alternative
predictions of effect concentrations (ECxIA / ECxDA), both maximum and minimum, that may
occur with any possible combination of toxicants that are included in the sample data set. In
case of 106 toxicants, these are more than 1.3 ● 1030 different combinations with 2 to 106
components. With the concentration ratio of mixture components, the ratio between
predictions (ECxIA / ECxDA) varies within a limited range. The deterministic simulation
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
results in an exact definition of these limits for any possible number of mixture components
and for any preset effect level x. However, when all dose or concentration response functions
of individual substances are described by using the Weibull model, these limits become even
independent of the magnitude of the effect level x.
For this simulation step, we therefore fitted all data sets to the Weibull model. The resulting
values for the slope parameter β varied between β = 1.6 and β = 8.9 for all chemicals but one
outlier, which had an extremely steep curve with β = 14.3. A graphical impression of the
meaning of these figures can be obtained by comparison with Figure 6, which shows Weibull
sample curves with slope values between β = 1 and β = 10. In the sample data set, the mean
was β = 3.7 and 80 % of the values exceeded β = ln 10 ≈ 2,3. This means, that for most of the
possible combinations of these toxicants, DA can be expected to predict a higher mixture
toxicity than IA.
Feeding these slope data into the computer simulations of maximum possible prediction
differences yielded the following results (Figure 9):
•
For any mixture that could be generated from the specific set of 106 chemicals the
corresponding predictions of effect concentrations by IA and DA will never differ by
more than a factor of 8.3, which is considerably smaller than the theoretical maximum
value of 106.
• If mixtures are composed only from those of the 106 chemicals which have the
concentration response curves with the lowest gradient, then situations occur, where
IA predicts a higher mixture toxicity than DA. However, the ratio ECxIA / ECxDA will
in no case drop below a value of 0.4, which means that the effect concentration
predicted by IA will never be smaller than 40 % of the corresponding value predicted
by DA.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Endpoint: Algal Reproduction
IA
ECxmix / ECxmix
CA
106
Any Mixture
20
10
4
Any Mixture that can be generated
from a specific set of 106 toxicants:
Maximum ratio = 8.3
Minimum ratio = 0.4
2
1
0.5
2
20
40
60
80
106
number of mixture components
Figure 9: Potential range of ratios between predictions of effect concentrations for the
algal toxicity of mixtures that could be generated from a data set for 106
chemicals. Note that ratios between predictions ECxIA / ECxCA are plotted on
a logarithmic scale.
13.4.4.2.Probabilistic simulations of distributions of prediction differences
The results of the deterministic simulation studies are in agreement with empirical mixture
studies and hence support the hypothesis that quantitative differences between IA and DA
predictions are typically relatively small, at least in comparison to other uncertainties in
regulatory risk assessments.
However, these results are based on the use of a non-threshold regression model, and might
therefore be misleading with respect to mixture risk assessment for non-genotoxic chemicals.
For instance, if all compounds in the mixture are present at doses below their thresholds, but
are estimated to be still effective, then IA erroneously predicts a mixture effect although no
one should be expected. Hence, the question arises whether and to what extent the
consideration of dose thresholds may affect the validity of the assumption that small
quantitative differences between IA and DA predictions are the rule.
In order to verify or falsify the validity of this assumption we performed probabilistic
simulations studies on the expected differences between IA and DA with the same data set. In
contrast to the deterministic approach, these simulations did not focus on the determination of
those mixture compositions that result in maximum differences between DA and IA
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
predictions, but possible dose ratios were considered evenly. This was done to gain insight
into the actual likelihood of significant over- or under-estimations that may result from the
assumption of DA in cases where in fact independent joint action occurs.
We investigated four scenarios with mixtures composed of 10, 20, 50 and 100 compounds,
respectively. In a first step we re-analysed the toxicity data for all 106 chemicals by adopting
the best-fit approach (Scholze et al. 2001) to a class of five non-linear regression models
(weibull, logit, probit, generalized logit I and II) that were extended to include a threshold
parameter. In short, all models were separately fitted to every data set, then for every
chemical the best fitting model was selected according to statistical criteria, and only this one
was then used for the subsequent simulation studies. In the second step, a hypothetical
mixture was simulated with compounds sampled by random (without replacement) from the
pool of 106 chemicals, and with mixture ratios also generated by random, so that every
possible mixture composition had an equal chance to occur (toxic units were generated from
the uniform distribution on the interval [0,1] and normalised in such a way that the sum of the
components’ fractions was one). Finally, the corresponding ratios between effect
concentrations predicted by DA and IA were calculated on the basis of the best-fit curves
derived from the threshold models. This simulation step was repeated 10.000 times in order to
generate distributions of the ratios between predictions.
For comparison, this procedure was finally performed a second time, by not using the
threshold versions of the models, but the original versions which assume continuous dose
response curves.
As a result, we obtained probabilistic information about the likelihood of over-estimations (or
under-estimations) of algal toxicity by the assumption of DA in situations of independent
joint action for four different mixture scenarios and two different approaches to low-effect
modelling. These distributions are depicted in Figure 10 and provide the following evidence:
•
It is confirmed, that DA usually provides the more conservative mixture toxicity
estimate. The likelihood for the reverse situation, i.e. that IA predicts a higher joint
toxicity than DA, is very low and tends to zero with increasing numbers of mixture
components.
•
After 10.000 simulations we never observed a ratio between the two predictions of
mixture-EC50 values that was greater than 4.2. This is considerably lower than the
maximum possible value of 8.3 that was determined for this data set in the preceding
deterministic simulation (see section 13.4.4.1 above). The difference results from the
fact that all possible mixture compositions had the same chance to be included in the
10.000 simulations, and it thus demonstrates how unlikely the occurrence of the
possible extreme case is. An enormously high number of simulations would have
obviously been required to cover also the highest possible values.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 10: Probabilistic simulations on predictions of algal toxicity of mixtures with 10,
20, 50 and 100 components, randomly composed from a data set of 106
different chemicals. Shown are the distributions of ratios between DA- and
IA-based predictions of EC50 values of the mixtures, resulting from 10.000
simulations. Experimental data sets for individual compounds were either
fitted to continuous regression models (black) or threshold models (red).
Dotted vertical lines refer to the 95% percentiles of the distributions. Figures
in the upper left corners provide the percentages of simulations in which IA
estimated a higher joint toxicity than DA (ratio <1).
•
As expected, the analysis of data sets by means of threshold-models leads to higher
differences between the alternative predictions than the use of non-threshold models.
In fact, this can be attributed to a resulting increase of the EC50 values of the mixtures
that are predicted under the assumption of IA, because the corresponding predictions
on the basis DA were unaffected by the inclusion of a threshold parameter in the
regression modeling (data not shown). The differences between the outcomes of both
modeling approaches increase with increasing numbers of compounds. However, even
with 100 mixture components, they did not differ by more than a factor of 2.2.
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Combined actions of chemicals in food through dissimilar modes of action
In additional simulation studies we also compared predictions for EC10 values of the
mixtures instead of EC50 values. The results were similar (data not shown).
13.4.5. Conclusions
•
Existing experimental evidences, mathematical analyses and results from simulation
studies give reasons to assume that quantitative differences between predictions of
effect doses or concentrations for multi-component mixtures derived from the
alternative models of independent action and dose additivity are relatively small for
realistic assessment situations, not exceeding an order of magnitude, and typically
only differing by a factor of less than 5 for mixtures with up to 100 components.
•
Evidence for the validity of this assumption mostly results from ecotoxicological test
data. There are reasons to assume that the principle also holds with data from
mammalian toxicology, but further research on this point is required.
•
Thus, in general the current status of knowledge about quantitative prediction
differences supports the use of dose additivity as a pragmatic and precautious default
approach to the predictive hazard assessment of chemical mixtures. Any lower
protection level should be justified by specific toxicological knowledge about the
mixture of concern.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.5.
Toxicological interactions
Toxicological interactions have been defined as “any toxic responses that are greater than or
less than what is observed under an assumption of additivity.”(EPA 2000). In this context,
‘additivity’ is typically the most appropriate concept from a choice of DA or IA. Interactions
that results in an effect less than expected under additivity are referred to as antagonism, subadditivity or inhibition, and those resulting in an effect greater than predicted are described as
synergy or supra-additivity.
Consideration of interactions is important because the use of component-based approaches,
such as DA or IA, assumes the absence of interactions. A very relevant question is how much
the occurrence of an interaction could alter the mixture effect from that predicted under
additivity. Of most concern is a possible synergy, or supra-additive interaction, that would
increase the level of toxicity compared to that expected. The literature relating to this question
has been critically reviewed (Boobis et al. 2011). Boobis et al. reviewed the experimental
evidence for synergies at low doses (defined as doses close to points of departure for
individual chemicals) in mixture studies and identified 90 studies, of which only 11 studies
reported a quantitative estimate of a low-dose synergy. Three criteria were identified to make
the quantification of synergy more consistent:
•
Synergy should be defined as departure from the mixture prediction using DA
•
A uniform procedure should be developed/used to assess synergy at low doses
•
The method used to define the POD used to assess synergy should be standardised.
Only 6 studies were considered to provide a useful quantitative estimate of synergy and these
comprised three studies of binary mixtures, two studies of five component mixtures and one
study of an 18 component mixture. When the magnitude of synergy was calculated based on
the ratio of observed to predicted dose for a fixed response (“Method A”) or the ratio of
observed to predicted response for a fixed dose (“Method B”) it was found that the magnitude
of synergy at low doses did not exceed that of the prediction made using DA by greater than a
factor of 4. The number of studies identified was not great enough to allow comparison of the
effect of using method A or B on the observed synergy.
Boobis et al. noted that the role of interactions is the subject of continued debate amongst
scientists and risk assessors, and that there is incomplete agreement on the impact of
interactions following exposure to a chemical mixture. Given the results of their review,
Boobis et al. considered that “there is probably merit in the default regulatory approaches
that assume toxicological interactions are not likely to occur at the low dose permitted under
existing exposure standards” however they acknowledged that this could not be a firm
conclusion, especially for the effects of cumulative and low-level chronic exposure, and that,
although the magnitude of observed synergies appears to be low, more work is required to
determine the frequency of synergy in real world situations (Boobis et al. 2011).
The presence of significant, unpredictable synergies could have questioned the use of any
additivity concept, including DA, as a default in cumulative risk assessment. However if
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issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
significant synergies can be considered unlikely, as is indicated, then the suitability of DA as
a conservative default is unaffected.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.6.
Approaches to cumulative risk assessment methods
The evaluation of experimental data describing the combined effects of chemicals, in this
report referred to as mixture effect assessment, has to be distinguished clearly from
approaches employed for conducting cumulative risk assessment in practice, here termed
cumulative risk assessment methods.
The application of cumulative risk assessment methods requires clarity about the goal of the
assessment. The aim can be to arrive at a risk estimate, an estimation of safe levels, of
margins of exposure, or can consist of ways of prioritizing certain mixtures, for further study
or for regulatory interventions. Estimations of safe levels or margins of exposure may be
based on worst-case-assumptions, but the prioritization of mixtures (or affected sites) has to
rely on fairly accurate quantitations of risk.
Almost all cumulative risk assessment methods in current use are applications of the concept
of dose addition. These include the Hazard Index (HI), Toxic Unit Summation (TUS), Point
of Departure Index (PODI), Relative Potency Factors and the TEQ concept.
Methods explicitly derived from independent action are not developed. An implicit
application of independent action is the assumption that mixture effects will not arise when all
chemicals in question are present at levels below their ADIs, with the additional implicit
assumption that ADIs represent true zero effect levels. It should be emphasised that the
implicit application of independent action can only be used for chemicals for which ADIs
have been derived. However, this is only the case for a small minority of chemicals in current
use.
13.6.1. Approaches based on dose addition (DA)
13.6.1.1.Hazard Index
The Hazard Index (HI) (Teuschler and Hertzberg 1995) is a regulatory approach to
component-based mixture risk assessment derived from DA and which can be generally
defined by the formula
n
HI = ∑
i =1
ELi
ALi
where EL is the exposure level, AL is the acceptable level, and n is the number of chemicals
in the mixture. Various measures for exposure levels and expectable levels may be applied;
the only constraint is that EL and AL must be expressed in the same unit. Input values for AL
can be ADIs or reference doses (RfD) for specific endpoints.
If HI > 1, the total concentration (or dose) of mixture components exceeds the level
considered to be acceptable. The method offers flexibility in applying different UFs when
defining AL for the individual substances.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
An assumption implicit in the use of the HI approach, and one that derives from the principles
of the DA concept, is that the acceptable levels AL for each individual chemical represent
exposures associated with the same (small or negligible) effect. In most cases, this is not
proven in practice, and will remain unproven in the foreseeable future. For most practical
applications, however, the error in making this assumption can be considered small.
13.6.1.2.Toxic Unit Summation
The method of Toxic Unit Summation (TUS) (Sprague 1970) is a direct application of the DA
concept and defined by the formula
n
n
i =1
i =1
TUS = ∑ TU i = ∑
ci
ECxi
where ci are the actual concentrations (or doses) of the individual substances in a mixture and
ECxi denote equi-effective concentrations (or doses) of these substances if present singly (e.g.
EC50i). The quotients ci / ECxi are termed Toxic Units (TU). Toxic Units rescale absolute
concentrations (or doses) of substances to their different individual toxic potencies. They
express the concentrations (or doses) of mixture components as fractions of equi-effective
individual concentrations (or doses) ECxi. Typically, x = 50 % (EC50i) is chosen as the
reference level, but TUS can also be calculated for any other effect level x. If TUS = 1, the
mixture is expected to elicit the total effect x. If the sum of Toxic Units is smaller or larger
than 1, the mixture is expected to elicit effects smaller or larger than x, respectively.
13.6.1.3.Point of Departure Index
The Point of Departure Index (PODI) is an approach to component-based mixture risk
assessment which is similar to the HI and TUS. In contrast to the HI, however, exposure
levels (EL) of chemicals in a mixture are not expressed as fractions of individually acceptable
levels (AL) but as fractions of their respective points of departure (PODs) such as NOAELs
or benchmark concentrations or doses (BML). In this way, different uncertainty factors that
may be included in AL values (see HI) are removed from the calculation (Wilkinson et al.
2000):
n
PODI = ∑
i =1
ELi
PODi
A PODI lends itself to the estimation of margins of exposure for the mixture of interest.
Similar to the HI, there is the implicit assumption that all PODs are associated with the same
effect magnitude, a principle derived from the features of DA.
13.6.1.4.Relative Potency Factors
The Relative Potency Factor (RPF) approach is another application of the DA concept for
mixtures of chemical substances that are assumed to be toxicologically similar (EPA 2000).
The concentrations (or doses) of mixture components are scaled relatively to the
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
concentration (or dose) of an index compound, and then summed up. The scaling factor is
called RPF. The total toxicity of the mixture is assessed in terms of the toxicity of an
equivalent concentration of the index compound. In general, the mixture concentration Cm
expressed in terms of the index compound for n compounds is
n
C m = ∑ (ci ∗ RPF i )
i =1
where ci is the concentration of the ith mixture component, and RPF1 = 1, as i = 1 indicates the
index chemical.
13.6.1.5.Toxic Equivalency Factors
The Toxic Equivalence Factor (TEF) is a specific type of RPF formed through a scientific
consensus procedure (EPA 2000). Based on the assumptions of a similar mechanism of action
of structurally related chemicals and parallel concentration (or dose) response curves, they
were first developed for dioxins. The total toxicity of the mixture is assessed in terms of the
toxicity of an equivalent concentration of an index compound. The total equivalent quantity
TEQ is estimated by summation of the concentrations (or doses) of mixture components ci
multiplied by the respective TEFi:
n
TEQ = ∑ (ci ∗ TEF i )
i =1
13.6.1.6.Data requirements and applicability of the cumulative risk assessment methods
All of the above cumulative risk assessment methods require at least rudimentary doseresponse information of individual mixture components which is used to derive the input
values, be they ADIs, RfDs, POD or information about relative potencies such as RPF or
TEF. Information about exposures must also be available.
The HI sums up ratios of exposure levels and ADIs or RfDs over chemicals. These estimates
can be arrived at by utilizing different uncertainty factors (UF) for each mixture component,
in order to deal with differences in data quality and sources of uncertainty.
If this is perceived to be inadequate, the PODI method can be used. PODI is based not on
reference doses, but on points of departure (NOAELs, benchmark doses). Extrapolation issues
(e.g. animal to human) are dealt with either by using one overall UF, or by estimating margins
of exposure.
The TEQ concept is predicated on the choice of a reference chemical and requires parallel
dose-response curves for all components. Both these requirements are often not met by
chemicals, but the method has been validated for dioxins and dioxin-like substances.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.6.2.
Approaches based on independent action (IA)
In general, CRA approaches based on IA are much less available than approaches based on
DA, see section 13.6.2. Approaches such as the Hazard Index or Toxic Equivalency Factors
do not have counterparts founded on IA principles and, because of the difference in
formulation of DA and IA, similar approaches may not be conceivable.
One pragmatic application of IA is the stance that a mixture effect will not occur if each
component is present at or below its individual zero effect level. However, this rests on the
use of true zero effect levels, whose identification may be controversial, and should not be
applied when effects are present but cannot be measured (when they are below the statistical
detection limits of the assay). This issue is discussed in detail in sections 13.7 and 13.8, and
has been thoroughly reviewed (EC 2009).
Simplified IA approaches that are sometimes mooted are the notions that 1) the mixture effect
is equal to the effect of the most potent component or that 2) the mixture effect is equal to the
summation of the effects of the components. As discussed in sections 13.1.4 and 13.6.2, these
approaches appear to rely on assumptions about the correlation of susceptibility to the
mixture components, these assumptions being rarely stated and potentially hard to
substantiate.
There would appear to be no practical approach for the use of IA in CRA, other than the
assumption that mixture effects will not occur if the individual components are without effect.
As discussed here and in section 13.8, this may not be a reasonable assumption.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.6.3. Use of the TTC in CRA approaches
13.6.3.1.The TTC concept
The TTC represents a level of human intake or exposure to a chemical that is considered to be
of negligible risk despite the absence of any chemical-specific toxicity data (Munro et al.
2008). Use of the TTC is an approach to risk characterisation that balances the uncertainties
in using data for other compounds against low levels of exposure. The TTC approach is not
intended for chemicals with an established risk assessment procedure, such as food additives,
pesticides and therapeutic drugs (Munro et al. 2008).
The TTC was initially developed by the FDA for packaging migrants and used a single value,
the threshold of regulation, of 1.5 µg/day. The rationale for this was summarised by Munro et
al.; briefly, this is the level at which most carcinogens have a less than one in a million
lifetime risk, and is 200-2000 times lower than the level at which other toxic effects occur
(Munro et al. 2008). Subsequently TTCs were set by dividing the 5th percentile of the
NOAEL distribution for e.g. Cramer class I chemicals, by what was termed “the usual 100fold uncertainty factor” and multiplied by 60Kg. Use of the 5th percentile gives a 95% chance
that the NOAEL for an unknown is higher than the NOAEL assumed in the TTC. Three
structural classes (see Table 11) of chemical were assigned TTC values of 1,800, 540 and 90
µg/day, followed by a TTC of 18µg/day for organophosphates and 0.15µg/day for chemicals
with structural alerts for genotoxicity.
TTCs for Cramer classes were originally intended for food flavours, the toxicities of other
chemicals may have different NOAEL distributions that affect the intended precautionary
value of the TTCs if this is not adjusted for. The separate TTCs for organophosphates and
genotoxic structural alerts reflect this. Compounds for which TTCs are not appropriate
include proteins, non-essential metals compounds and dioxin-like chemicals (Munro et al.
2008). The TTC approach may need modification (i.e. a modified input distribution of
NOAELs or modified conversion process from NOAEL to TTC) for chemicals with specific
routes of exposure (e.g. topical application of cosmetics), unless it is validated for these
situations.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 11: TTC values
TTC, µg/person/day
1800
Cramer class
Cramer class I
540
Cramer class II
90
Cramer class III
18
0.15
Description
Simple structures, efficiently metabolised, low
potential toxicity
Less clearly innocuous than class I, no positive
indications of toxicity, no lack of data for toxicity
Structural features that preclude a strong initial
presumption of safety, or that indicate toxicity
Organophosphates
Structural alerts for genotoxicity
A recent EFSA Opinion suggested that the TTC value for Cramer class II structures was not
well supported by the underlying toxicological data, and proposed that class II structures
should be classified as if they were Class III structures (EFSA 2011a). The Opinion listed
categories of substances for which the TTC approach should not be used as:
•
High potency carcinogens (i.e. aflatoxin-like, azoxy- or N-nitroso-compounds)
•
organic substances
•
Metals
•
Proteins
•
Substances that are known or predicted to bioaccumulate
•
Substances with structures that are not adequately represented in the original
databases from which the TTC values have been derived, e.g. nanomaterials and
radioactive substances
•
Substances likely to have the potential for local effects on the gastro-intestinal tract
13.6.3.2.Use of the TTC in cumulative risk assessment
A concept such as the TTC may be valuable in cumulative risk assessment since in most cases
there will be a lack of toxicological data for some of the mixture components. The concept
may allow such gaps to be bridged with conservative assumptions, rather than causing an
assessment to founder on a lack of data.
Price et al. explored the use of the TTC in estimating mixture toxicity, and concluded that use
of the TTC led to conservative estimates that could be suitable for screening mixture
assessments (Price et al. 2009). Price et al. found that the use of the TTC could overestimate
the toxicity of a mixture if it was applied to a mixture component that was the dominant
contributor of the mixture risk, but not if it was applied to a component that was not a
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issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
dominant component. Consequently it was suggested that the impact of using the TTC should
be evaluated in each case, and if the overall risk estimate was driven by components for
which TTC values were used then the estimate should be taken with caution (Price et al.
2009).
In the unlikely, or perhaps undesirable, event that the TTC approach is used for all
components in a mixture, then risk could still be deemed acceptable on this basis, providing
exposure levels are low, but an indication of risk would indicate the need for refinement
(rather than immediate concern or action). In this case refinement would mean the collection
of toxicological data to replace the TTC. If a mixture assessment based on the TTC is used to
prioritise components for data collection, then this is equivalent to prioritising on exposure
levels (if all the TTC classifications are the same) or on exposure levels weighted by
structural features (if more than one TTC classification is included).
The use of TTCs in a mixture assessment is explored further in case study 3 (section
15.8.3.4).
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Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.7.
Cumulative risk from sub-ADI levels
A key reference point in single substances’ and mixture toxicology is the “acceptable daily
intake“(ADI). ADI stands “for an estimate of the amount of a substance in food expressed on
a body weight basis that can be ingested daily over a lifetime, without appreciable risk to any
consumer on the basis of all known facts at the time of evaluation, taking into account
sensitive groups within the population (e.g., children and the unborn).” (Regulation (EC) No
396/2005). Other reference values, such as the “tolerable daily intake” (TDI) from JECFA or
the “reference dose” from U.S.EPA (RfD) are similarly defined. ADIs are derived by
considering one substance at a time.
It is frequently argued, most recently in the opinion of the three EU Scientific Committees
SSCP, SCHER and SCENIHR on Toxicity and Assessment of Chemical Mixtures (DG
Health and Consumer Protection 2011), that combination effects are not to be expected for
dissimilarly acting substances when each chemical is present at levels around its ADI. The
Committees stated that:
“The TDIs, DNELs or equivalent values are expected to represent a value at which no effects
are produced; thus for threshold substances, the assumption is that this value is equal to or
lower than the no-effect level; thus an E(Ci)=0 should be assumed for exposures at the TDI
or DNEL level. Consequently, the co-exposure to several substances all below the estimated
TDI, DNEL or equivalent value should be assumed to be negligible if all substances have
dissimilar modes of action.” (emphasis added)
As summarized in section 13.1.3, under IA mixture effects can be ruled out with certainty
only if all components are present at doses equivalent to “zero-effect levels”. If, however,
some compounds in the mixture produce small, but statistically not significant effects, joint
effects may cumulate to significant levels, particularly when large numbers of chemicals are
present that all affect a common endpoint.
It is obvious that the principles of IA place a heavy burden on the ability to distinguish zero
effect levels from small effects that may be impossible to verify experimentally.
Thus, in connection with ADIs, three specific questions arise from the viewpoint of mixture
risk assessment and mixture toxicology:
•
•
Is it reasonable to associate an ADI with a “zero-effect level” in all cases?
Are there effects below ADIs, which have no impact on health after exposure to single
substances but may be relevant in the case of exposure to mixtures?
• Is it valid to extrapolate observations made with single substances or their mixtures in
the dose range equivalent to “no observed adverse effect levels” (NOAELs) to arrive
at risk estimates regarding sub-ADI exposure levels?
These considerations are of principal interest with regard to the assessment of mixture
toxicity:
•
The principles of mixture toxicity (DA, IA or interaction) are generic and apply to all
endpoints, not only to adverse health effects. Therefore, the question of possible
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Combined actions of chemicals in food through dissimilar modes of action
effects at levels below ADIs, here referred to as “sub-ADI effects”, has to be extended
to non-adverse effects.
A discussion of these issues is presented in the following sections. We will first deal with the
question as to whether ADIs and TDIs can with certainty be associated with zero-effect
levels, before turning to the issue of effects at sub-ADI levels, so-called precursor effects.
13.7.1. Extrapolation from NOAEL or Benchmark Dose to ADI
13.7.1.1.10 x 10 Assessment Factors
To derive an ADI, it is customary to use a NOAEL from experimental animal studies for
endpoints representative of long-term toxicity. The NOAEL is divided by an assessment
factor (also referred to as an uncertainty factor) to arrive at an ADI. A default assessment
factor of 100 is normally used. Instead of NOAELs, benchmark doses can also form the basis
of ADIs. In this case, the lower confidence limit of a benchmark dose is combined with an
assessment factor.
The default factor of 100 that is used in these assessments is the composite of two separate
assessment factors:
•
•
a factor of 10 for interspecies extrapolation, and
a factor of 10 for intraspecies extrapolation.
For the purposes of this report, we disregard further assessment factors for “LOAEL to
NOAEL” extrapolations, acute to chronic or for “route-to-route” extrapolation. EFSA
recently initiated a project to re-evaluate and, potentially update, these assessment factors in
the future. In specific cases, where data of high quality are present, assessment factors with
lower numerical values can be used, although this has only been done in rare instances, as
evidenced by numerous PRAPeR documents.
NOAELs are not necessarily doses without adverse health effects. Especially because of the
small number of animals used per dose group and the resulting limited statistical power,
NOAEL may be associated with effects. EFSA have stated: “The size of the effect at the
NOAEL is, on average, over a number of studies, close to 10% (quantal responses) or 5%
(continuous responses)”(EFSA 2009a).
13.7.1.2.Interspecies Assessment Factor
The interspecies assessment factor of 10 is the composite of two factors, one for allometric
scaling to take account of different body sizes between species, the other to adjust for
differences in toxicokinetics. For allometric scaling from the rat to other species, a factor of 4
is generally used, and this is well supported by numerous evaluations (Kalberlah and
Schneider 1998;Schneider et al. 2004). If the experimental studies are performed with mice, a
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issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
factor of 7 would be more appropriate. Conversely, a smaller factor of 1.7 is justified in the
case of studies with dogs.
Interspecies differences are not always fully addressed by the allometric scaling factor.
Although justified in most cases, there may be deviations (towards smaller or larger species
differences) due to toxicokinetic and/or toxicodynamic reasons. For this reason, an additional
factor of 2.5 is introduced, which, however, is less well supported by experimental evidence.
It may have been chosen for reasons of convenience: the product of 4 x 2.5 yields the
traditional interspecies factor of 10.
13.7.1.3.Intraspecies Assessment Factor
To cover differences in sensitivity between human subjects, an intraspecies variability factor
of 10 is used. There is only limited empirical evidence to support the numerical value of this
factor.
WHO proposed to separate this factor into a toxicokinetic and a toxicodynamic component,
with two subfactors of 3.2 (WHO 1999).
Although not demonstrated on the basis of large datasets, it is generally assumed that among
healthy human subjects there is greater inter-individual variation in susceptibility towards
toxic substances than in experimental animals. The reason for this is that highly inbred,
genetically homogenous strains of laboratory animals (mostly rodent species) are used for
toxicological investigations, and these strains show a considerably smaller variability in
susceptibility than human populations.
A second reason for the higher inter-individual variability in susceptibility among humans is
the presence of vulnerable groups in the general population such as the elderly, children
(although for certain effects children should not generally be considered to be more
susceptible than adults), or individuals with compromised health.
The quantitative assessment of this variability is complicated. Hattis and colleagues have
compiled a large database of individual studies, which investigated differences in effectrelated observations or toxicokinetic parameters in humans (Hattis et al. 1999a;Hattis et al.
1999b;Hattis and Anderson 1999). As a first attempt to use such data to put extrapolations on
a sound footing, the database was utilized by Schneider et al. to derive distributions to be used
in a probabilistic model for effect assessment (Schneider et al. 2006). For the data sets about
differences in effect-related doses in adults retrieved from the Hattis database these
distributions describe the ratios between the group mean effective dose and the dose causing
the effect in a specified (low) quantile of the examined population.
The following ratios of the medians of distributions were obtained by Schneider et al. from
the Hattis-database (Schneider et al. 2006):
•
•
•
dose ratio average susceptible (median) / 10% quantile of groups investigated: 3.31
dose ratio average susceptible (median) / 5% quantile of groups investigated: 4.82
dose ratio average susceptible (median) / 1% quantile of groups investigated: 9.96.
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Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
This should be considered a proof of concept – further efforts are needed to substantiate the
numerical values by expanding and differentiating the database.
WHO recommended that the intraspecies factor of 10, if possible, should be replaced by a
data-derived factor for a particular substance (WHO 1994). It is further mentioned that in
specific cases this factor of 10 might not be sufficiently protective. Reference is made to the
case of genetic polymorphisms, which may cause large differences in the internal dose
between population subgroups that carry different alleles of enzymes responsible for
xenobiotic metabolism.
In a recent paper, Dorne reviewed data on the inter-individual variability due to toxicokinetic
factors (but omitting considerations of toxicodynamic differences between individuals which
should also be taken into account) (Dorne 2010). When evaluating pharmacokinetic data the
highest variability was observed with substances which are metabolised via pathways
including polymorphically expressed xenobiotica-metabolising enzymes. According to this
analysis, factors of up to 4.7 (99th percentile, for CYP2D6-dependent pathways) would be
necessary to consider resulting differences in internal doses. Relevant variability could also
be caused by age (neonates with immature metabolic capacity for some specific pathways,
and elderly people).
In the subsequent paragraphs we discuss the level of protection which can be attributed to an
ADI or equivalent value. In this context, effect data which are measured on a continuous,
numerical scale (e.g. body weights, protein concentration in urine, etc.) have to be
distinguished from dichotomous variables (categorical or quantal) which provide information
about incidences, e.g. the number of affected subjects in a group of exposed individuals.
13.7.1.4.Continuous Responses
In the context of risk assessments, a certain level has to be defined as the threshold for
adversity for continuous effect variables (e.g. activity of transaminases in blood, protein
concentration in urine, etc.). These critical effect levels have to be determined individually for
each effect, as adversity depends on the nature of the effect, physiological background ranges,
etc.
In the hypothetical example in Figure 11 the critical effect level for excretion of protein x in
urine was determined as a urinary concentration of 5 µg/l. Higher urinary protein excretion
would be considered an adverse effect. The dose group showing a protein concentration of 5
µg/l (group mean) in urine was considered to represent the NOAEL. An ADI can be derived
by application of assessment factors with the implicit assumption that in susceptible humans
this concentration would also not be exceeded.
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Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 11: Schematic presentation of ADI derivation from continuous variables from
experimental animal data (dose-response curve on the right): POD is the dose
at which the critical effect size (here: 5 µg/l) is not exceeded. By application of
assessment factors this level of protection is transferred to the average human
population and the subpopulation of susceptible humans
Figure 12 translates this into a probability distribution showing the incidence for exceeding
the critical effect level of 5 µg/l in the general population. Theoretically, at each dose level a
distribution of urinary concentrations in the population can be assumed, with the percentage
lying above the critical effect level decreasing with decreasing dose. The percentage of the
population that exceeds this level is shown in Figure 12 as an incidence curve. In this figure
the assessment factors applied are large enough to ensure that also for the most susceptible
individuals the critical effect level is not exceeded at the ADI (incidence zero or close to zero
%).
In Figure 13 the population dose-response curve is moved to the left, with the consequence
that, starting from the POD, the assessment factors are not large enough to cover also the
most vulnerable individuals.
The dose-response curve in the general population is rarely known.
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Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 12: Schematic presentation of ADI derivation from continuously measured
experimental animal data (dose-response curve on the right): Hypothetical
case A: assessment factor sufficient to prevent exceeding a critical effect size
in susceptible individuals
Figure 13: Schematic presentation of ADI derivation from continuously measured
experimental animal data (dose-response curve on the right): Hypothetical
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
case B: assessment factor insufficient to prevent exceeding a critical effect
size in most susceptible individuals
With continuous data, an increase of the number of animals per group will not automatically
alter the group mean values measured. But larger experimental groups will have an influence
on the uncertainty of the measured values (e.g. expressed by the standard deviations to the
mean values). Thus, if the critical effect level is defined as absolute value (example:
concentration of methaemoglobin not to exceed 5%) larger experimental groups will reduce
the overall uncertainty of the measurement data, but will not tend to change the mean dose
designated as POD.
13.7.1.5.Quantal (Dichotomous) Responses
With quantal variables, the number of affected individuals within an exposure group is
counted. The aim is to identify a dose group in which the frequency of the affected animals is
significantly higher than with the next lower dose. The highest dose without significant
effects is called NOAEL. With results from a standard repeated-dose experiment (e.g. a 90day oral rodent study according to OECD Test Guideline 408, with 10 animals per sex and
dose group) it can be assumed that effects in the 10% incidence range might be detectable,
depending on the type of effect, background rate and other factors.
Alternatively, by dose-response modeling a dose associated with a pre-determined low effect
level (e.g. 10% incidence above background) can be estimated as benchmark dose (BMD; for
a 10% effect level: BMD10). The lower confidence limit of the BMD is called BMDL.
The following figures schematically present the ADI derivation for categorical data.
Figure 14 exemplifies the situation where assessment factors lead to an ADI below a
population threshold (incidence at ADI: 0%). In Figure 15 application of assessment factors
results in an ADI connected with a relevant incidence for the adverse effect under
investigation (approx. 5% in this hypothetical example).
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 14: Schematic presentation of ADI derivation for categorical experimental
animal data (dose-response curve on the right): Hypothetical case A:
assessment factor sufficient to include most susceptible individuals in general
population
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 15: Schematic presentation of ADI derivation for categorical experimental
animal data (dose-response curve on the right): Hypothetical case B:
assessment factor insufficient to include most susceptible individuals in
general population
As the dose-response curve of the human population is unknown, the actual level of
protection provided by the ADI is also indeterminate. If the assessment factors applied are
higher than that needed to take account of inter- and intraspecies differences, the
corresponding incidence level in the human population will be well below the 10% level (in
case of a BMD10 used as POD). If the NOAEL is associated with a lower effect size, the ADI
may well be equivalent to a zero effect level. In other cases, where the animal model used for
a specific substance and effect is insensitive (which might lead to overlooking an effect) or
where there is an especially sensitive population subgroup (e.g. with a yet unknown enzyme
polymorphism), then the effect incidence in the sensitive population group (or – depending on
the size of this sensitive group – in the general population) may even be higher than that
observed in the experimental animal group at the POD.
It should be mentioned that in the case of categorical data, those animals showing effects at
the POD will be the more susceptible ones. Even when it is agreed that the variability in inbred experimental animals is much lower than that between human individuals it must be
recognised that by applying an intraspecies factor to a dose derived from a categorical
NOAEL, intraspecies variability is counted twice
At the lower end of the dose-response curve the uncertainty in determining the dose-response
relationship increases sharply (due to the intrinsic difficulty to determine low-level effects),
with the consequence that even for the best investigated substances the dose-response
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issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
relationship in this area and the existence or non-existence of population thresholds becomes
indeterminate.
Primarily for gross apical endpoints, thresholds (i.e. dose level associated with 0 increased
incidence) might also be observed at a population level. But in many cases even for
substances with an assumed threshold, due to poorly understood toxicity mechanisms and
lacking information on inter-individual differences (which, in case of a threshold mechanism
means lacking knowledge on individual thresholds) dose- and effect estimates in the low-dose
area will become uncertain.
13.7.1.6.Conclusion
The quantification of residual risks associated with ADIs, especially for sensitive subgroups
in the general population, is impossible. Major uncertainties arise from the (generally
unknown) inter-individual variability in the human population.
The assumption that the usually applied assessment factor of 100 sufficiently accounts for
interspecies differences in vulnerability, as well as intraspecies variability to yield zero effect
levels in all cases is not supported by empirical evidence, and largely unverifiable.
The uncertainties associated with ADIs and TDIs are recognized by risk assessors. The three
EU Scientific Committees (DG Health and Consumer Protection 2011) pointed out that “there
is obvious uncertainty in setting the TDI or DNEL.” Furthermore, risk assessors do not
exclude some residual health risk if exposures to doses equivalent to ADIs occur. There is
regulatory guidance which advises as follows: “when the critical effect is judged of particular
significance, such as developmental neurotoxic or immunotoxic effects, an increased margin
of safety shall be considered” (Regulation (EC) No 1107/2009).
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.8.
The issue of non-adverse effects at levels below ADIs
This section deals with evidence for first biological changes that might occur at dosages
below ADIs. Such effects are not necessarily to be classed as adverse, but they too might sum
up to significant levels when there is exposure to multiple chemicals, with consequences that
are largely unexplored.
The following aspects are of relevance:
•
•
•
•
13.8.1.
Effects below a “critical effect size” defined to separate adverse responses from those
assumed to be of no harm, for continuous response variables.
Issues arising from the dichotomisation of continuous variables into quantal variables
that define individuals as “affected” or “not affected”.
The possibility that upstream events leading to overt disease might cumulate to
produce adverse effects, although the upstream events themselves, so-called precursor
effects, may not be adverse.
Biochemical alterations that occur in low dose ranges.
Effects below a critical effect size
From a biological point of view, an effect may be irrelevant even though it is not strictly zero.
Biological systems are capable of correcting certain disturbances provoked by exposure to
chemicals, however the challenge lies in establishing a relevant effect size that defines the
borderline between “effect” and “no-effect” and “adversity“ and “lack of harm“ in a
biological or toxicological, and not a statistical sense.
Several approaches exist to defining an effect of relevance quantitatively. In criterionreferenced evaluations, the importance of an effect is judged in relation to a clear biological
or clinical criterion. An example would be the procedure for defining a critical sperm count
below which fertility experts recommend assisted fertilisation. This is based on information
about correlations between sperm count and fertilisation success in human populations. The
criterion used was the point below which fertilisation rates began to decrease with lower
sperm counts (Jorgensen et al. 2006).
In many cases, however, straightforward biological or clinical criteria are not available and in
these situations effects of relevance are defined by norm-referenced evaluations. This
involves establishing a critical effect by considering the variance of the effect parameter in
the population under investigation. There are numerous examples that follow this approach.
Cut-off points for elevated cholesterol levels, low birth weights or late onset of walking in
children are all derived by determining certain percentiles – often the 95th - of cumulative
population frequency distributions of the selected effect variable. In mutagenicity testing,
critical mutation frequencies are defined in terms of multiples of standard deviations of
background mutation rates.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
If a “critical effect size” is used for deriving an ADI by defining a dose threshold for the
experimental data there will always be a “sub-critical” effect size above zero, which is classed
as not adverse and which is associated with doses below the chosen NOAEL or BMDL.
A pertinent example is the critical effect size used by the Dutch RIVM for white blood cell
counts during the assessment of mycotoxins (T-2/HT-2). Reductions of white blood cell
counts of more than 5% were classed as critical. Conversely, reductions in counts of less than
4% are interpreted as non-adverse effects, and doses associated with these smaller effects are
assessed as presenting no concern. Similarly, decreases of acetylcholine esterase activity of
more than 20% are regarded as adverse (Bokkers et al. 2009).
While these procedures may make sense when exposure to single chemicals is considered,
problems might occur with exposures to multiple chemicals that also affect the endpoint
under investigation. In such cases, effects below the critical effect size may well sum up to a
combination effects above the cut-off effect sizes used for the assessment of single chemicals.
13.8.2. Issues arising from the dichotomization of continuous effect variables
Issues equivalent to those discussed in connection with critical effect sizes may arise when
continuous effect variables are dichotomized to classify individuals as “affected” or “not
affected”, and when dose thresholds are subsequently defined on the basis of these
classifications. In such cases, biochemical or precursor effects that might be present in
individuals assigned as “not affect” might also build up to effects above the line regarded as
“affected”, when exposure is to multiple chemicals.
13.8.3. Precursor Effects
Biological events that precede overt adverse outcomes measured in chronic toxicity assays are
often referred to as precursor effects. With regard to certain disease states, the scientific
understanding of the molecular events, precursor effects, that advance pathological processes
to a level where they become manifest as disease or adverse effect has improved
considerably. It has been recognized that there is great potential in using these precursor
effects, instead of adverse outcomes themselves, as tools for the screening of chemicals
(Woodruff et al. 2008). In certain instances, it has been suggested to regard the precursor
effects themselves as adverse. They occur at doses lower than those required to elicit overt
effects in long-term animal studies. Examples for this stem from advancing knowledge about
thyroid disruption, and from disruption of the function of male sex hormones.
A prominent example is thyroid disruption. It is well established that thyroid hormone
insufficiency during pregnancy can cause lasting neuro-developmental deficits in the affected
child. Thyroid hormone levels are determined through a complex interplay between several
factors, including uptake of iodine from dietary sources which is required for the synthesis of
the hormone, transport of iodine to the thyroid gland, synthesis of the precursor of active
thyroid hormone, transport of the precursor to tissues and deiodinisation of the precursor to
the active thyroid hormone. Feedback processes exist to ensure that declining thyroid
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
hormone levels in the blood are compensated through increased synthesis in the thyroid
gland. Thyroid hormone levels can serve as an early indicator of downstream adverse effects
on neurodevelopment, and it has been suggested to regard diminutions of thyroid hormone as
an adverse effect in itself, although it is distinct from the induction of overt
neurodevelopmental effects (Woodruff et al. 2008).
Suppression of thyroid hormone levels in experimental animals occur at lower doses than
those necessary to produce overt toxicity. For example, perchlorate inhibits iodide uptake in
the thyroid at rather low concentrations. Much higher levels of perchlorate are required to
induce adverse effects on development or reproduction, secondary to a decline in serum
thyroid hormone levels and an increase in thyroid stimulating hormone. This may lead to
thyroid hypertrophy, hypothyroidism and finally to reproductive (prenatal or postnatal)
effects. However, compensatory mechanisms may prevent an effect on circulating hormone
levels. The NAS stated that one would need a 75% inhibition in iodide uptake for the
perchlorate effect to be adverse (Murray 2005).
There is good evidence that several chemicals can work together to induce reductions in
thyroid hormone levels at doses where the individual chemicals are ineffective (Crofton et al.
2005). (Crofton 2008) demonstrated that chemical substances may interact with the thyroid
hormone pathways at different sites and via different mechanisms.
Woodruff et al. reported from a workshop in 2007, that the participants agreed that early
biological perturbations and precursor effects (like hormonal changes) should be regarded as
adverse outcomes in experimental studies (Woodruff et al. 2008). However, in current risk
assessment practice, usually only the overt outcome, but not the precursor effect, is
considered adverse, and the ADI (or TDI/RfD) is usually derived from such a late
“downstream end point”. To illustrate the point, Woodruff et al. demonstrated a continuum
between an adverse and a non-adverse outcome with precursor effects which may occur well
below the apparent adverse effect “threshold”, which is the ADI/TDI after (interspecies- and
intraspecies-) transformation as shown in Figure 16 (Woodruff et al. 2008).
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 16: Distribution of a typical physiological parameter within the population and
how that may vary depending on the influence of chemical and biological
background (adopted from Woodruff et al.(Woodruff et al. 2008) and
reprinted with kind permission of the editor)
13.8.4. Biochemical Alterations
Biochemical changes are specific precursor effects at the molecular level. A “no observed
effect level” for many biochemical changes may be well below the doses required for the
induction of adverse effects which are used to derive ADIs. For example, the “no observed
transcription effect level” (NOTEL) is frequently observed well below NOAELs for toxicity.
Even if some substances in a mixture act via dissimilar modes of action, some of the isolated
biochemical changes may be identical at very low concentrations.
Zarbl et al. described the NOTEL as a much more sensitive indicator of (not necessarily
adverse) effects and demonstrated the relationship in a schematic way (see Figure 17) (Zarbl
et al. 2010).
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Linear extrapolation
e
s
n
o
p
s
e
R
NOAEL
NOEL
NOTEL
Experimentally observed
Log dose
Figure 17: Hypothetical dose-response curve as applied to risk assessment. The solid line
indicates a typical response curve that shows a threshold for the
experimentally measured endpoint. These data are then used to determine a
NOAEL and/or a NOAEL. Data are then used to determine a reference
dose…. The use of genomics provides the opportunity to define much more
sensitive NOTEL (indicated on graph), NOAEL or a mechanistically based
benchmark dose … for use in risk assessment”. Citation from, and graph
redrawn from, (Zarbl et al. 2010).
On the basis of empirical data it is not always straightforward to demonstrate relevant
biochemical changes at very low concentrations below a NOAEL or even below an ADI.
Below, some examples from in vivo and in vitro studies are provided.
Zheng et al. studied the hepatotoxic potential of four different substances with regard to
biochemical marker proteins and gene expression (Zheng et al. 2011). The substances acted
through different modes of toxic action. Dose ranges up to 1000-fold below those needed to
affect conventional toxicity endpoints were chosen for this test. Effects on changes of gene
expression of cytochrome P450 enzymes were observed.
Judson et al. analysed perturbations of a biological pathway as an early event leading to
adverse (apical effects) typically observed at higher concentrations, which they referred to as
a “biological pathway altering dose” (BPAD) (Judson et al. 2011). Below a BPAD an
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
adaptive effect is assumed. Although derived from in vitro-studies results were transformed
into an in vivo-dose estimate for precursor effects also covering variability and uncertainty of
this estimate. Figure 18 (adopted from this publication) clearly points to BPADs well below
the apical No Effect Level (NEL) and the Low Effect Level (LEL) for liver hypertrophy. The
estimated BPADLs (red circles) are located below the NEL/100 (red triangle), which would
correspond to an ADI (NEL/100), for 7 out of 14 examples for pesticides (conazoles).
In the case of cumulative exposures some substances may well lead to identical perturbations
of pathways which might affect apical endpoints if critical levels are reached.
These examples provide additional indications for the possibility of effects below ADI for the
single substances (in this case: pesticides), which should be considered in the assessment of
combined exposures to mixtures. There are further examples of biochemical changes at very
low concentrations (Abril et al. 2011;Ludwig et al. 2011;Schulpen et al. 2011;Thomas et al.
2011;Zheng et al. 2011).
Figure 18: Comparison of “high throughput chemical risk assessments with LEL and
NEL values for liver hypertrophy from animal studies on the 14 conazole
fungicides in Phase 1 of ToxCast. …For each chemical, the black box gives
the population-variability-derived (1%,99%) confidence intervals about the
median BPAD. The whiskers indicate uncertainty-derived 95% confidence
intervals about the extremes of the variability confidence interval. The
BPADL99 is indicated by a red circle: the LEL by a blue box: the NEL by a
grey triangle and the NEL/100 by a red triangle. Estimated chronic exposure
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
levels from food residues are indicated by vertical red lines. All values are in
mg/kg/day” (cited from Judson et al. (2011); for further explanation see
description in text). Reprinted with permission from (Judson et al. 2011)
Copyright (2011) American Chemical Society.
13.8.5. Conclusions
The observations and issues presented in this chapter show that biochemical changes and
precursor effects have been reported in dose ranges below those associated with ADIs or
TDIs. Together with the uncertainties discussed in section 13.7, these reports cast further
doubt on the notion that ADIs are true zero effects levels in all cases.
The uncertainty is introduced because the dose-response relationship is rarely known for an
adverse effect in the sensitive human population. This means that the ADI may not be a zeroadverse effect level for the critical effect observed above NOAEL. Extrapolations to the subADI range do not necessarily mean a principle change from observations at/above NOAELlevel or a switch from ‘high dose’ to ‘low dose’ considerations and assumptions.
The issues examined in this chapter support the possibility of mixture effects including those
driven by IA at a sub-ADI exposure level. The principles of IA may also hold true for
nonadverse effects, which may cumulate to exceed homoeostasis and become adverse in case
of exposure to mixtures.
In conclusion, simultaneous exposure to multiple chemicals may result in cumulative effects
even when the chemicals are present at levels below ADIs.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.9.
Data requirements in legislation and availability of such data
For (cumulative) risk assessment both hazard assessment and exposure data are needed. The
complexity and quality of these data determines the quality of the risk assessment. A
comprehensive legal framework exists in the EU which sets out the data requirements for
toxicological and exposure data for substances in food. The following section summarises
which data are legally required for hazard and exposure assessment in the EU and discusses
the consequences for risk assessment.
13.9.1. Toxicology data available to regulators
Food, whether from vegetable or animal origin, may contain ingredients, added intentionally
or unintentionally, apart from its intrinsic constituent parts. It is EFSAs remit to perform
scientific risk assessment for food and feed which form the basis for risk management
decisions which should protect consumers from health risks from the food chain. In the
following section we will analyse the possible differences in the amount and quality of data
available on health risks associated with intentionally or unintentionally added food
ingredients.
Ingredients added intentionally to food should improve its properties such as taste, odour,
stability or impart a certain property to food. These food improvement agents are
•
food additives
•
food flavourings
•
food enzymes
Their use is only permitted as far as no health risk is associated with their use or a special
beneficial effect is achieved.
As with food, some substances, so called feed additives, are added intentionally also to feed
to improve its properties. These might be consumed by humans who eat food from animal
origin.
Furthermore, substances may be added for nutritional purposes like vitamins, minerals or
certain amino acids in e.g. foods for particular nutritional uses or as ingredients in food
supplements.
Another group of food ingredients are substances which are used intentionally during the
production or packaging of food. Their presence in food is usually not wanted but cannot
totally be avoided. Therefore, concentration limits which should be without appreciable risk
for the consumers have been established for these substances
•
food contact materials
•
residues of plant protection products
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
•
residues of veterinary drugs
•
residues of biocidal products used in animal husbandry
Other substances may be detectable in food or feed, although their presence is not wanted but
cannot be avoided due to e.g. their overall presence in the environment:
•
food contaminants
•
feed contaminants (“undesirable substances in animal nutrition”)
13.9.1.1. Legal requirements for the provision of toxicity data for food improvement agents
Food additives, food enzymes and food flavourings make up the group of the so called food
improvement agents. A common authorisation procedure has been laid down for these food
improvement agents. A central part of the authorisation process is the safety evaluation of
these substances to show that these substances do not pose a safety concern to consumers.
The legal framework encompasses Commission Regulation (EU) No 234/2011 (EU, 2011)
and Regulation (EC) No 1331/2008 (EU, 2008a) on a common procedure for evaluation and
authorization of these substances, Regulation (EC) No 1332/2008 (EU, 2008g) on food
enzymes, Regulation (EC) No 1333/2008 (EU, 2008f) on food additives, and Regulation (EC)
No 1334/2008 (EU, 2008e) flavourings.
Specific data requirements for risk assessment of food additives, food enzymes and
flavourings are listed in Commission Regulation (EU) No 234/2011 and are summarised in
the table below.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 12: Specific data requirements (core data set) for risk assessment of food
improvement agents
Food additives
Toxicokinetics
Subchronic toxicity
Genotoxicity
Chronic
toxicity
carcinogenicity
Reproductive
developmental toxicity
Food enzymes
Food flavourings
examination for structural/metabolic
similarity to flavouring substances in an
existing flavouring group evaluation
(FGE)
Subchronic toxicity
Genotoxicity b
Subchronic toxicity a
Genotoxicity b
Chronic toxicity and carcinogenicity a
/
and
Developmental toxicity a
a: where applicable
b: specified in the guidance (see below): information on the ability to induce gene mutations and structural and numerical
chromosomal aberrations
Additional guidance on the evaluation of the food improvement agents are presented in three
guidance documents (EFSA, 2009a; b; 2010). The guidance on food additives from 2009,
which is currently under re-evaluation (finalisation announced for summer 2011), does not
provide a specification of the requirements on toxicological data additionally to those
requirements already addressed in the regulation. But in the former guidance of the Scientific
Committee on Food (Guidance on submission for food additive evaluations;
http://ec.europa.eu/food/fs/sc/scf/out98_en.pdf) a detailed list of toxicological data necessary
for the evaluation of food additives was presented:
metabolism / toxicokinetics
subchronic toxicity
genotoxicity (gene mutations in bacteria, gene mutations in mammalian cells in vitro,
(preferably the mouse lymphoma tk assay), induction of chromosomal aberrations in
mammalian cells in vitro; positive results in any of the above in vitro tests will normally
require further assessment of genotoxicity in vivo.
chronic toxicity and carcinogenicity in two species
reproduction and developmental toxicity (at least a two generation study in one species and
developmental toxicity data in two species)
Additional studies (e.g. immunotoxicity, allergenicity, food intolerance, neurotoxicity, human
volunteer studies, in vitro studies as alternatives to in vivo studies, special studies, acute
toxicity, skin and eye irritation and skin sensitisation) may be indicated if necessary.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The guidance on flavourings specified the needs for data on genotoxicity (information on the
ability to induce gene mutations and structural and numerical chromosomal aberrations in
vitro), developmental toxicity (a developmental study in rodents) and repeated dose toxicity
(a 90-day feeding study in rodents, preferably in rats). Further toxicity tests may be requested
by the Panel if a need for additional testing would arise from the submitted data. Also the
guidance on enzymes specified the needs for data on genotoxicity (information on the ability
to induce gene mutations and structural and numerical chromosomal aberrations in vitro,
verification of positive results in vitro with in vivo tests) and an oral subchronic toxicity test.
Additional studies to the core set may be requested. But it might also be possible to reduce or
completely wave toxicological test, e.g. in case the safe use of the enzymes is documented.
In summary, comprehensive toxicity data have to be submitted in the context of the
authorization process of food additives. For food additives data on toxicokinetics, subchronic
and chronic toxicity have to be generated as well as data on specific endpoints like
mutagenicity, carcinogenicity or reproductive toxicity. Usually information on the NOAEL
and LOAEL for all these endpoints will be available from these studies. The core set of
toxicological requirements for food enzymes and food flavourings encompasses much less
studies. Only if there are concerns about the toxicological hazards of these substances more
data may be requested. This highlights the difficulties in cumulative risk assessment. If for
example the chronic cumulative exposure to some food additives and some flavourings should
be assessed, there might be the difficulty that for the food additives NOAEL-values from
chronic studies are available, but only NOAEL from sub-chronic studies may be available for
the flavourings. Due to the different study designs, these NOAEL values are not directly
comparable to each other. For further discussion see section 13.9.1.9.
13.9.1.2. Legal requirements for the provision of toxicity data for substances for nutritional
purposes
This group encompasses substances added for nutritional purposes in
•
foods for particular nutritional uses,
•
fortified foods or as
•
ingredients in food supplements.
Food for particular nutritional uses is used as a synonym for “dietetic foods” or “dietary
foods”, i.e. these are foodstuffs for e.g. infants or young children in good health, or for
persons whose digestive processes or metabolism are disturbed (Directive 2009/39/EC (EU,
2009)).
“Fortified foods” means food to which micronutrients like vitamins or minerals have been
added. The legal framework for addition of vitamins, minerals or certain other substances is
described in Regulation (EC) No 1925/2006 (EU, 2006a).
Food Supplements are defined as „concentrated sources of nutrients or other substances with
a nutritional or physiological effect whose purpose is to supplement the normal diet. They are
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
marketed 'in dose' form i.e. as pills, tablets, capsules, liquids in measured doses etc.”
(http://ec.europa.eu/food/food/labellingnutrition/supplements/index_en.htm).
“Nutrients”
stands for vitamins and minerals (Directive 2002/46/EC (EU, 2002b)).
Activities of EFSA have mainly concentrated on the regulation of minerals and vitamins. At
the moment, the substances that can be added for nutritional purposes are controlled through
positive lists (see “Administrative guidance on submission for safety evaluation of substances
added for specific nutritional purposes in the manufacture of foods”
(http://ec.europa.eu/food/food/labellingnutrition/nutritional/adm_guidance_safety_substances
_en.pdf). Before any substance is added to such a positive list an application for authorization
has to be performed.
Thus far, no formal procedure for the toxicological risk assessment of these substances for
nutritional purposes has been published. For vitamins and minerals present in food
supplements upper safe limits for the daily intake as well as minimum amounts per daily
portion of consumption have to be established. Furthermore, reference intakes for the
population (so called recommended daily allowances; Directive 2008/100/EC (EU, 2008d))
are requested (Article 5 of Directive 2002/46/EC). The maximum amounts should be based on
scientific risk assessment based on generally accepted scientific data. Regarding the scientific
opinions for the vitamins and minerals it becomes obvious that toxicological effects have
been investigated comprehensively, i.e. whole data set from acute to chronic toxicity as well
as reproductive toxicity are available for nearly all substances under consideration.
13.9.1.3. Legal requirements for the provision of toxicity data for food contact materials
Food contact materials are defined as materials and articles intended to come into contact
with foods such as packaging materials, cutlery and dishes, processing machines, containers,
materials and articles in contact with water for human consumption. The legislative
framework for food contact materials is laid down in Regulation (EC) No 1935/2004 (EU,
2004a). Food contact materials should under normal or foreseeable conditions of use not
transfer their constituents to food in quantities which could endanger human health.
According to Regulation (EC) No 1935/2004 Article 5 an authorisation procedure is required
for:
•
substances for use in food contact materials and articles, especially substances used in
plastic materials and articles, substances used for active or intelligent functions in
active and intelligent materials, and substances used in regenerated cellulose films
•
materials and the articles themselves
•
manufacturing processes of food contact materials and articles specifically chemically
recycling processes for plastics to be used in plastic food contact materials
Toxicological requirements during the authorisation process are described in the “Guidance
document on the submission of a dossier on a substance to be used in Food Contact Materials
for evaluation by EFSA by the Panel on additives, flavourings, processing aids and materials
in contact with food (AFC)” (http://www.efsa.europa.eu/en/efsajournal/doc/21r.pdf). The
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
amount of toxicological data which have to be submitted for authorisation depends on the
quantity of the substance which might migrate into the food:
•
high migration (5 - 60 mg/kg/food): extensive data set
•
migration between 0.05 – 5 mg/kg food: reduced data set
•
low migration (<0.05 mg/kg food): limited data set
“Core extensive data set”
- 3 mutagenicity studies in vitro
i) test for induction of gene mutations in bacteria
ii) test for induction of gene mutations in mammalian cells in vitro (preferably the
mouse lymphoma tk assay)
iii) test for induction of chromosomal aberrations in mammalian
cells in vitro
- 90-day oral toxicity studies, normally in two species
- studies on absorption, distribution, metabolism and excretion
- studies on reproduction in one species, and developmental toxicity,
normally in two species
- studies on long-term toxicity/carcinogenicity, normally in two species
“Reduced core set”
- 3 mutagenicity tests
- a 90-day oral toxicity study
- Data to demonstrate the absence of potential for accumulation in man
“Limited data set”
- 3 mutagenicity tests
Additional studies or special investigations may be necessary in case if prior knowledge or
structural considerations reveals that these studies are indicated.
To prevent a risk to human health migration limits have been established for some
components of food contact materials. For example for plastic materials an overall migration
limit of 10 mg of substances/dm2 of the food contact surface for all substances that can
migrate from food contact materials to foods exists. For individual authorized substances
specific migration limits (SML) have been fixed on the basis of a toxicological evaluation, i.e.
with regard to the ADI or TDI of a specific substance.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.9.1.4. Legal requirements for the provision of toxicity data for novel foods and novel food
ingredients
Novel foods and novel food ingredients are foods and food ingredients that have not been
used for human consumption to a significant degree in the EU before 15 May 1997
(http://ec.europa.eu/food/food/biotechnology/novelfood/index_en.htm). The legal background
is laid down in Regulation (EC) No 258/97. Novel foods and novel food ingredients have to
be notified before they can be placed onto the market. A risk assessment has to be performed
for the notification. If there are any concerns or objections an authorisation process is
initiated. Currently there are no obligatory recommendations on the toxicological data to be
submitted for the risk assessment (see Recommendation 97/618/EC (EU, 1997b)).
13.9.1.5. Legal requirements for the provision of toxicity data for residues
Food from animal or plant origin may contain residues of
•
plant protection products
•
pharmacological active ingredients including pharmacological active biocidal
products used in animal husbandry.
Those residues should not be harmful to the consumer. Plant protection products,
pharmacological active ingredients and biocidal products have to be authorised before they
can be placed on the market. Comprehensive toxicological investigations and risk assessment
is a central part of the authorisation procedure. If necessary, maximum residue levels (MRL)
are established or the application of the substances is prohibited. According to Annex II of
Council Directive 91/414/EEC (EU, 1991) the following toxicological tests are stipulated for
the authorisation procedure of plant protection products: data on
•
metabolism and toxicokinetics
•
acute toxicity (at least after oral and dermal exposure)
•
skin and eye irritation
•
skin sensitisation
•
toxicity after repeated exposure (90-day study in rats and dogs)
•
genotoxicity (gene mutation in bacteria, in vitro mammalian cell gene test, in vitro
mammalian cytogenicity test plus additionally in vivo testing if necessary)
•
long term oral toxicity in the rat
•
carcinogenicity test in the rat and mouse
•
two generation study in rats
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
•
study on developmental toxicity in rats and rabbits
•
additional studies e.g. delayed neurotoxicity can be performed if necessary
Testing procedures for pharmacological active ingredients and biocidal products are
comparable to those of plant protection products, i.e. comprehensive toxicological data are
available for these groups but are not reported here in detail.
13.9.1.6.Legal requirements for the provision of toxicity data for contaminants
Food contaminants have not been added intentionally to food. These substances, like
mycotoxins, heavy metals, polycyclic aromatic hydrocarbons or acrylamide may be present in
food as a consequence of the production procedure, storage conditions, transport or
packaging. Another large group of contaminants cannot be avoided in food due to their
overall presence in the environment. Food contaminants shall be kept as low as can be
reasonably achieved following good working practices. Maximum levels have been set for
certain
food
contaminants
in
order
to
protect
human
health
(http://ec.europa.eu/food/food/chemicalsafety/contaminants/index_en.htm). These maximum
levels have been set on basis of toxicological risk assessments as well as considering socioeconomic and risk management considerations. There are no formal requirements for the
toxicological investigations to be performed. But, for most of these substances large numbers
of toxicological studies exist.
If sufficient toxicological data are available Tolerable Weekly Intakes (TWI) are derived for
food contaminants. The procedure for derivation of TWIs is basically comparable to the
procedure for setting Acceptable Daily Intakes (ADI). These values are expressed as weekly
intake, because the daily intake may show great variations in contrast to the weekly intake. To
indicate that contaminants are not really wanted in food the guidance values are called
“tolerable” instead of “acceptable”.
13.9.1.7. Legal requirements for the provision of toxicity data for undesirable substances in
animal nutrition –
Undesirable substances in animal nutrition are the counterparts to food contaminants.
Undesirable substances are “any substance or product … in and/or on the product intended for
animal feed which presents a potential danger to human health, animal health or the
environment
or
do
not
adversely
affect
livestock
production”
(http://ec.europa.eu/food/food/animalnutrition/contaminants/index_en.htm). Like for food
contaminants maximum limits for e.g. heavy metals, dioxin, aflatoxin, certain pesticides have
been established for feed materials, feed additives and feedingstuffs (see Directive
2002/32/EC (EU, 2002a) and its amendments Commission Directive 2003/57/EC (EU,
2003b) and Commission Directive 2003/100/EC (EU, 2003c)).
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.9.1.8. Legal requirements for the provision of toxicity data for feed additives
Feed additives should improve the quality of feed and the quality of food from animal origin,
or
improve
the
animals’
performance
and
health
(http://ec.europa.eu/food/food/animalnutrition/feedadditives/index_en.htm). Feed additives
have been classified in the following categories:
•
technological additives (e.g. preservatives, antioxidants, emulsifiers)
•
sensory additives (e.g. flavours, colorants)
•
nutritional additives (e.g. vitamins, minerals, amino acids, trace elements)
•
zootechnical additives (e.g. digestibility enhancers, gut flora stabilizers)
•
coccidostats and histomonostats.
The rules for authorisation, supervision and labeling of feed additives are set out in
Regulation (EC) No 1831/2003 (EU, 2003a).
Detailed rules for the implementation of Regulation (EC) No 1831/2003 of the European
Parliament and of the Council are laid down in Commission Regulation (EC) No 429/2008
(EU, 2008b). As a basic requirement for the authorisation of feed additives studies concerning
the safety of its use for the target animal and for the consumer are required. Based on the
provided studies the safety for the consumer shall be evaluated and potential residues of the
additive or its metabolites in food derived from animals given feed or water containing or
treated with the additive shall be established. The aim of the safety evaluation is to establish
an ADI. Toxicological data generally required are guideline studies on
•
toxicokinetics;
•
acute toxicity;
•
genotoxicity (mutagenicity, clastogenicity; at least gene mutations in bacteria and/or
in mammalian cells, induction of chromosomal aberrations in mammalian cells, in
vivo test in mammalian species);
•
sub-chronic oral toxicity;
•
chronic oral toxicity/carcinogenicity;
•
reproduction toxicity including
developmental toxicity study);
teratogenicity
(two
generation
study
and
This set of toxicological studies can be supplemented by other studies e.g. on immunotoxicity
or neurotoxicity as far as necessary.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.9.1.9. Summary and conclusions
The legal data requirements for the different substance groups are summarised in the
following table.
Table 13: Summary of the legal requirements for toxicological data for different
substances added to food in the European Union (further details are
presented in the text)
Toxicokinetics/metabolism
Acute toxicity
Skin and eye irritation
Skin sensitisation
Genotoxicity a
Subchronic toxicity
Chronic
toxicity/carcinogenicity
Reproductive
toxicity,
endpoint fertility
Reproductive
toxicity,
endpoint developmental
toxicity
Additional studies
X
X
X
X
X
X
X
X
X
X
X
X
X
(XX
)
(XX
)
X
X
(XX
)
(X)
(X)
X
X
(X)
(X)
(X)
X
X
X
X
X
(X)
X
X
X
(X)
X
X
(XX
)
(XX
)
(XX
)
X
X
X
(XX
)
X
X
X
X
X
X
X
X
(XX
)
(X
X)
(X): test not formally required but available in most cases
(XX): further tests may be requested if scientifically justified
a: usually a core set of in vitro tests is required which may be supplemented with additional tests if necessary
b: no formal procedure for the toxicological risk assessment have been published
c: substance not added intentionally to food or feed, no formal requirements on toxicological data
d: “high migration”: 5 - 60 mg/kg/food; „medium migration: 0.05 – 5 mg/kg food; „low migration: < 0.05 mg/kg food
There is a substantial difference between substances added intentionally or unintentionally to
food or feed. Substances added intentionally to food or feed have to fulfill certain defined
data requirements which should enable the authority to assess the potential toxicological risk
associated with the uptake of these substances. Besides the core set of data additional data can
be requested in case there are any concerns about the safety of these substances.
Supporting publications 2012:EN-232
Residues from plant
protection products
Novel foods and novel
food ingredients b
Food contact materials
with low migration d
Food contact materials
with medium
Food contact materials
with high migration d
Undesirable substances
in animal nutrition c
Food contaminants c
Food flavourings
Food enzymes
Food additives
Feed additives
Study type
Substances for
nutritional purposes b
Substance type
126
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Data requirements may differ substantially between the different substance groups. Complete
data sets are required for example for pesticides, nearly complete datasets for food and feed
additives and food contact materials with high migration. But only rudimental toxicological
data may be available for example for food contact materials with low migration if there are
no further concerns.
Up to now formal data requirements are missing for novel food and novel food ingredients.
There are also no formal data requirements for food or feed contaminants, because their
presence in food or feed is not wanted but cannot be avoided e.g. due to the overall presence
of in the environment or their formation during the manufacturing process. But there is no
manufacturer or importer who is responsible for these substances and who can be forced to
perform toxicological studies.
These potential differences in data availability may limit the options of cumulative risk
assessment of chemical substances in food. Cumulative risk assessment may be performed
with respect to acute or chronic toxicity or regarding certain endpoints like reproductive
toxicity. At a higher CRA tier, endpoint specific points of departure (e.g. NOAELs, BMDLs)
may be used. Table 13 obviously demonstrates the limits of such an approach. Based on the
legal requirements it cannot be expected that for all substances present in food comparable
toxicity data are available. For example, data on acute or chronic toxicity are not requested
for all substance groups, even data on subchronic toxicity are not available for all groups.
The problem for the risk assessor arising from this situation may be that no information on the
NOAEL or LOAEL for a certain endpoint such as hepatotoxicity is available or that only
information from toxicity studies with a different design is available, e.g.: For substance A a
NOAEL for hepatotoxicity from a chronic toxicity study with rats is available and for
substance B a NOAEL (or even only a LOAEL) from a subchronic study with dogs is
available. For certain contaminants it is even likely that there are human toxicity data which
have to be compared to toxic effects in experimental animals. The NOAELs or LOAELs from
these different studies are not directly comparable to each other, and this may compromise
efforts to aggregate such heterogeneous data by using methods for cumulative risk
assessment. It would need further expert judgement to transfer these data by regarding species
differences and differences in exposure time to comparable reference values.
This illustrates the basic difficulties in cumulative risk assessment. Comparable data (e.g.
NOAELs from toxicity studies with identical design) should be available for compounds from
one substance group. But even this is not guaranteed, because certain studies have only be
performed if there is a specific concern. But such studies are not legally required, which
means that even within one substance group the quality and nature of the available data may
be very heterogeneous.
13.9.2. Exposure data available to regulators
Exposure assessment is the second important factor besides hazard characterization for risk
assessment. Generally, exposure via diet (“dietary exposure” or “dietary intake”) can be
calculated by multiplication of the concentration of a chemical in food with the food
consumption (WHO, 2009).
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Dietary exposure =
As in hazard assessment an acute and a chronic dietary exposure can be calculated, depending
on whether an acute or chronic risk shall be evaluated. In the following section we will
discuss the basic principles and the legal background for dietary exposure assessment mainly
focusing on pesticide regulations. The other sections will only shortly be addressed.
13.9.2.1. Legal requirements for the provision of exposure data for pesticide residues
Obviously, for a detailed exposure assessment sufficient and qualified data on the
concentration of chemicals in food as well as on food consumption are necessary. The general
legal basis for the collection of food exposure data in the EU is laid down in Regulation (EC)
No 178/2002 (EU, 2002c). Regulation (EC) No 882/2004 (EU, 2004b) established the general
legal provisions for food inspections and monitoring on official controls performed to ensure
the verification of compliance with feed and food law, animal health and animal welfare.
Regulation (EC) No 882/2004 applies to all food ingredients.
In the context of plant protection products the preparation of annual reports on pesticide
residues is based on the legal provisions laid down in Regulation (EC) No 396/2005 (EU,
2005), Chapter 5: “Member states are required to establish national control programmes and
to carry out regular official controls on pesticide residues in food commodities in order to
check compliance with the MRL for pesticide residues and to assess the consumers exposure.
The information has to be submitted to the European Commission, to EFSA and the other
Member States”.
Apart from the national control/monitoring programmes (designed by each country) a second
programme, a coordinate European programme, has been established (Regulation (EC) No
1213/2008 (EU, 2008c)) for pesticide residues. It is the aim of the EU coordinated
programme to provide statistically representative data regarding pesticide residues in food
available to European consumers. In November 2011 the “2009 EU Report on Pesticide
Residues” has been published by EFSA (EFSA, 2011). The report delivers a summary of the
national monitoring data and inter alia
•
•
•
•
an analysis of the results of the monitoring data on pesticide residues provided;
some ideas about the reasons why certain MRLs were exceeded;
an analysis of chronic and acute risks to the health of consumers from pesticide
residues;
an assessment of consumer exposure to pesticide residues based on the information
provided under the first bullet point and any other relevant information available,
including reports submitted under Directive 96/23/EC.
In 2009 ten food commodities have been investigated in the EU coordinated programme. In a
three year cycle investigating about 10 different commodities every year, the major
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
components of the European diet, which are represented by 20-30 products, can be
investigated by this procedure (the commodities analysed in the three year cycle represent
between 39% and 95% of the diets of the different EU nations). These data will provide a
representative basis for the estimation of the exposure to pesticide residues in food of
European consumers (EFSA, 2011). In 2009 food from plant origin was analysed for 120
different pesticides (100 of them mandatory) and food from animal origin for 32 pesticides
(29 of them mandatory).
For risk assessment purposes the data on pesticide residues in the different food commodities
were combined with the data on food consumption as presented in the EFSA Pesticide
Residue Intake Model (PRIMo). The consumption data of PRIMo are based e.g. on national
food surveys which collected long term and short term consumption data for the whole
population or more differentiated for different age groups. Apart from the national food
consumption data also data from the WHO cluster diets are incorporated in PRIMo which
allows the consideration of different food patterns in the individual EU member states or a
calculation for larger than national groups.
Food consumption data for the EU member states are also available from the “Concise
European Food Consumption Database in Exposure Assessment”
(http://www.efsa.europa.eu/en/datexfoodcdb/datexfooddb.htm). However, this database only
provides consumption data on a limited number (15) of broad food categories, which are only
suitable for a preliminary exposure assessment.
Apart from PRIMo which is based on national and international data on food consumption,
national databases and models also exist. For example, the German Federal Institute for Risk
Assessment (BfR) first developed the VELS model in 2005, which is based on the
consumption data of children aged 2-5 years, which are considered to be the most sensitive
subgroup due to the relative high food intake in comparison to body weight. In 2011 a new
model for the calculation of dietary exposure has been established, which also includes the
food consumption data of adults (aged 14-80 years) and the subgroup of women in the child
bearing age (aged 14-50 years). These data are based on the results of a national food survey
performed in 2006 (BfR, 2011). In the Netherlands a probabilistic model has been developed
to calculate the acute or chronic intake of pesticide residues, the so called MCRA (Monte
Carlo Risk Assessment; https://mcra.rivm.nl/) (Pieters et al., 2005). Probabilistic models are
also available from Ireland (CREMe 2), United Kingdom (CSL), the University of Bremen,
Germany, and the USA (DEEM/Calendex, CARES, Lifeline and SHEDS) (EFSA, 2008).
Discussion
Monitoring programmes deliver numerous data on pesticide residue concentrations in food.
Apart from the data from “surveillance samples” which are collected without any particular
suspicion there are also “enforcement samples”. These are taken if there is suspicion about the
safety on non-compliance with the legal limits (i.e. these data are not representative for the
food available on the EU market). They are collected as part of the national food monitoring
programmes and compiled by EFSA for the EU member states. In 2009 about 120 pesticides
have been monitored in 10 commodities in the EU coordinated programme. Within the 3 year
circle of the EU coordinated programme data on the most relevant 30 food commodities are
collected. The EU coordinated and national data on chemical concentrations in food are
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
combined with food consumption data using excellent models which reflect different food
patterns from several European countries. Thus, dietary intake of pesticide residues in the EU
is very well documented due to the legal requirements in the EU.
Nevertheless, there are some shortcomings in this system. Currently about 500 pesticides are
authorized in the EU, and probably 1000 worldwide (EFSA, 2011). Although the EU the
three-year-monitoring programme delivers an enormous data base, not all pesticides on the
market can be covered due to costs and efficiency reasons. Furthermore, not all food
commodities can be considered: MRLs have been set for about 400 food commodities. These
are partially covered by the national monitoring programmes, but data gaps still remain,
because not all residue data in one commodity can be transferred or extrapolated to another.
An additional difficulty in pesticide monitoring is that MRLs are established for Raw
Agricultural Commodities (RAC) of plant or animal origin. In some cases the MRL refer not
only to the edible parts of the plant but also comprise inedible parts (e.g. bananas with peel).
Food monitoring data are established in a way that the compliance with the legal requirements
can be checked. But this does not necessarily mean that these data deliver information of the
real pesticide residue concentration taken up by the processed food.
13.9.2.2. Legal requirements for the provision of exposure data for substances in live animals
and products of animal origin
The legal requirements for substances in live animals and products of animal origin are
comparable to those for pesticide residues: The general legal basis for the collection of food
exposure data is laid down in Regulation (EC) No 178/2002 and Regulation (EC) No
882/2004. The requirements for substance control planning for live animals or products of
animal origin are laid down in Council Directive 96/23/EC (EU, 1996). This is amended by
Commission Decision 97/747/EC (EU, 1997a) which establishes levels and frequencies of
sampling for certain animal products. As for pesticide residues, EFSA generates an annual
summary report on the results of residue monitoring in food of animal monitoring in the
Member States which comprises not only results on residues from veterinary products but
also other substances (e.g. pesticides) and contaminants (EC, 2010). The underlying data on
substance concentrations in the different food stuffs can together with the existing data on
food consumption be used for a calculation of the dietary intake of substances from live
animals and products of animal origin.
13.9.2.3.Legal requirements for the provision of exposure data for food improvement agents
Intake of food additives and flavourings has to be monitored by the member states which have
to report their findings to the Commission and Authority (Article 27 of Regulation (EC) No
1333/2008; Article 20 of Regulation (EC) No 1334/2008). There are no specific requirements
for enzymes regarding food monitoring.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.9.2.4. Legal requirements for the provision of exposure data for food supplements
Food supplements are provided in a dosed form which contains a recommendation for the
daily intake and a warning not to exceed the stated recommended dose. Effective monitoring
should be guaranteed (see Directive 2002/46/EC). Maximum levels and minimum effective
doses have also been set for vitamins and minerals added to foodstuff. But no monitoring
programme like for the other food ingredients has been established.
13.9.2.5. Legal requirements for the provision of exposure data for food contact materials
During e.g. processing, manufacturing, or packaging food comes into contact with numerous
materials (plastics, metals, ceramics, etc.). To ensure that these materials do no liberate
substances into food in an amount that would impair food safety official controls have to be
performed in order to perform compliance with Regulation (EC) No 1935/2004. The
European Reference Laboratory for Food Contact Materials (EURL-FCM) has been
established in 2003 which is responsible for the production and dissemination of
internationally accepted quality assurance tools, including validated methods, reference
materials, reference measurements, interlaboratory comparisons and training. The EURLFCM cooperates with a supporting network of European National Reference Laboratories.
13.9.2.6. Legal requirements for the provision of exposure data for food contaminants
Data on contaminant concentrations in food are collected in Europe on basis of Regulation
(EC) No 882/2004 and Regulation (EC) No 1881/2006 (EU, 2006b) which are amended by
several other regulations like e.g. Commission Regulation (EC) No 333/2007 (EU, 2007)
where the methods of sampling and analysis for certain food contaminants are described.
Coordinated data from Member States can be collected within a scientific cooperation
(SCOOP) task. The legal basis for this procedure is Council Directive 93/5/EEC (EC, 1993).
This directive lays down a procedure whereby Member States can focus their scientific
resources in a co-ordinated manner on problems in the area of food. Pooled data on particular
issues of concern regarding food safety can be collected for different substance groups.
Beyond these EU wide activities, the WHO Global Environment Monitoring System – Food
Contamination Monitoring and Assessment Programme (GEMS/Food) contributes to a
relevant amount to the measurement of contaminant concentrations in food worldwide. But
GEMS/Food also established important data on the food consumption worldwide
(GEMS/Food cluster diets).
13.9.2.7.Summary and conclusions
Monitoring programmes deliver numerous data on the concentration of pesticide residues,
food contaminants and other ingredients of food. Apart from the data from “surveillance
samples” which are collected without any particular suspicion, also “enforcement samples”
which are taken if there is suspicion about the safety on non-compliance with the legal limits
(i.e. these data are not representative for the food available on the EU market) are collected in
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
the food monitoring programmes. These data have to be submitted to EFSA and many of
these data are published by EFSA.
Apart from the data on substance concentrations in food in the last years several programmes
became publicly available which provide information on food consumption and can be used
for the calculation of dietary intake. While qualified monitoring data are necessary for a
skilled and realistic exposure assessment, the existing models also allow a worst case
consideration: Based on the existing food consumption data and the legal MRL for the
individual commodities maximal intakes can be calculated for a lower TIER in a tiered risk
assessment approach.
Although the existing monitoring system provides large datasets on dietary exposure there are
still some relevant shortcomings in the assessment of chemicals via dietary intake. For
example monitoring programmes can never encompass all substances possibly contained in
food and will never cover all food commodities. Data on substance concentrations as
collected by food monitoring must not necessarily represent the substance uptake by food due
to further processing of the food, i.e. they usually represent a worst case. This means that an
uncertainty analysis of the data, whether the calculated intake is rather an over- or
underestimation of the real intake, should be performed.
Today generally qualified monitoring data are available which can be used for a skilled
exposure assessment. But due to the fact that monitoring is always limited due to cost and
effectiveness reasons there will always be data gaps as not all substances potentially added to
food (un-intentionally or intentionally) can be monitored in all commodities.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
13.10.
Cumulative exposure assessment (CEA)
The data requirements for use of IA or DA were considered in the relevant sections on each
additivity concept; however these sections concentrated on the requirements on effect data,
rather than the requirements on the corresponding exposure data that is also required for a risk
assessment. We therefore considered whether the choice of IA or DA alters the data
requirements for cumulative exposure assessment (CEA) or if the exposure assessment is
essentially independent of the model. Consideration of the mathematical and practical
features of the concepts, sections 13.1 and 13.2, suggests that the latter situation applies, in
that a risk assessment using either IA or DA concepts requires exposure data but the choice of
model does not alter the type or amount of information required.
The availability of data for CEA is currently a major knowledge gap in that most studies do
not measure exposure to multiple chemicals simultaneously and therefore assumptions must
be made as to how the exposure to multiple chemicals should best be estimated when the
information for each chemical is derived from different samples and, often, different studies.
Knowledge of variations in cumulative exposure within populations is also lacking, for
example whether certain members or sections of the population are highly exposed to
multiple chemicals or if exposure is more balanced.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TASK 3: GROUPING CRITERIA
The literature areas relevant to this task were the definition of dissimilar action, mode and
mechanism of action; the literature was collected and summarised in Tasks 1 and 2, and
presented in sections 10.1, 11 and 13.3.
The insights from these activities are directly relevant to the proposed approach (Task 5) and,
to avoid duplication, further description has been incorporated into that section of this report
(section 15.7).
A 2008 report “Cumulative risk assessment for phthalates - the tasks ahead" (NRC 2008)
contains several insights into the grouping of chemicals with varying mechanism of action
which now are reviewed here.
The report noted that there are ambiguities in the criteria that are proposed for grouping
chemicals in CRA, and that the use of grouping criteria that are inappropriately narrow may
prevent chemicals that do indeed cause a common toxic effect from being assessed together.
The report strongly recommends a physiologically based approach to grouping in which
chemicals that cause any or all of the end effects attributed to a selected physiological process
would be grouped together (NRC 2008). For example, during development the action of
androgens drives male sexual differentiation, and this can be disrupted through a variety of
molecular mechanisms, such as androgen receptor antagonism and reduced testosterone
synthesis, producing end effects in males such as nipple retention and hypospadias. A
physiologically based approach would group all chemicals that caused either end effect,
nipple retention or hypospadias, together for assessment.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TASK 4: ASSESS APPROACHES TO CRA FOR DISSIMILARLY ACTING CHEMICALS, EVALUATE THE
DOSE ADDITIVITY APPROACH
We have assessed the available approaches and guidance for risk assessment of mixtures,
including European approaches ((EFSA 2008b;EFSA 2009c), American (EPA 1986;EPA
2000) and international (IPCS/WHO 2009;Meek et al. 2011) approaches. We have
concentrated on areas of the guidance relevant to a dissimilar mode of action. These
approaches are described below.
The use of DA as a conservative default assumption was reviewed by (EC 2009) and is
discussed in this report in sections 13.15, 13.4.1, 13.5 and 14.3. The situations where the DA
approach might not be sufficiently conservative, i.e. if IA was the more appropriate model
and DA under-predicted risk, were reviewed in section 13.4. To summarise, an experimental
situation in which DA produced a less conservative mixture effect prediction than IA was not
identified in our systematic literature search (section 13.1.5).
14.
Existing approaches to CRA for dissimilarly acting chemicals
Opinion of the Scientific Panel on Plant Protection products and their Residues to
evaluate the suitability of existing methodologies and, if appropriate, the identification
of new approaches to assess cumulative and synergistic risks from pesticides to human
health with a view to set MRLs for those pesticides in the frame of Regulation (EC)
396/2005 (EFSA 2008b).
The Panel limited their considerations to plant protection products for pragmatic reasons (the
general lack of data for other products) and also limited their considerations to the impact of
dose addition, deliberately omitting response addition (IA) and interactions.
The Panel proposed criteria for including chemicals in a common assessment group (CAG)
and highlighted the possibility of refining the group in a stepwise fashion. Criteria for
grouping included general criteria (chemical structure, mechanism of pesticidal action) or
more refined criteria (common toxic effect, toxic mode of action). The approaches considered
for CRA were, in order of increasing complexity and refinement,
•
the hazard index,
•
the reference point index,
•
the relative potency factor method and
•
physiological based toxicokinetic (PBTK) modelling.
Specific Panel recommendations of relevance to this project task were:
•
To use a tiered approach to toxicological evaluation and intake estimation
•
To evaluate assumptions and uncertainties at least qualitatively, with quantitative
examination of those factors potentially critical to the outcome
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
•
To increase the usefulness of exposure data for CRA by, for example, designing
monitoring programs and establishing reporting levels with the needs of CRA in
mind
Scientific Opinion on Risk Assessment for a Selected Group of Pesticides from the
Triazole Group to Test Possible Methodologies to Assess Cumulative Effects from
Exposure through Food from these Pesticides on Human Health (EFSA 2009c).
The 2009 Opinion carried out a cumulative risk assessment for triazole fungicides using the
methods proposed by the earlier opinion (EFSA 2008b).
As a result of this experience, the panel proposed a simplification of the tiered approach, as
follows:
•
From an early stage, the CAG should be as refined as the data allows.
•
Exposure assessments should be limited to one deterministic and one probabilistic tier
The panel concluded that “although a tiered approach is an appropriate way to address
cumulative dietary risk assessment it cannot yet be applied on a routine basis”
The PPR Panel identified the following issues:
•
the basis for and establishment of CAGs on a European level,
•
definition and agreement on desired levels of protection,
•
improvement of the robustness of methodologies of cumulative exposure assessment
and development of guidance on their appropriate use.
The panel also noted that:
•
“when assessment of a CAG based on relatively broad criteria, due the absence of
information on mode or mechanism of action for the common toxicological effect,
fails to give adequate reassurance, this may serve as a trigger for further research, to
enable the assessment to be completed.
•
The establishment of relevant CAGs is the starting point for all cumulative risk
assessments.
•
Consensus should be reached at an international level on the criteria and compounds
that should be used to create a CAG, to avoid differences between national cumulative
risk assessments.
•
An important issue is that a first tier should be more conservative compared to the
next tiers.
•
In itself, the hazard assessment tiers are clear and could be performed for any CAG.”
An important lesson from the case study could be that a variety of methods are now available
but that data on exposure and toxicological effects are not readily available for use with the
methods. The greater availability of data, on standardised platforms, would actually assist in
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
the comparison of methods, since there could be less focus on compromises that tolerate data
gaps and the risk estimates from different methods can only be compared when the method is
able to run through to completion.
EPA: Guidelines for the Health Risk Assessment of Chemical Mixtures (EPA, 1986) and
Supplementary Guidance (EPA, 2000).
EPA guidance has several areas of relevance. If a component based approach is selected, the
guidance indicates a choice between “toxicologically similar” mixtures, with assessment by
the hazard index or relative potency factor approaches, and “toxicologically independent”
mixtures to be assessed by response addition (i.e. IA). (Figure 2-1, EPA, 2000). The Guidance
notes that this choice may rely on data that is not readily available or may require scientific
judgement. It is also noted that “the true toxicologic mechanism of action...is rarely known for
a given mixture or even for most of its components” and “the judgements that are made of
toxicologically similar action or independence of action, for example, will be uncertain”. The
Guidance proposes that the assessor should deal with this by implementing several of the
assessment approaches, and evaluating the range of health risk estimates that are produced.
The Guidance also states that:
•
response addition (IA) is the default approach when “the component chemicals are
functionally independent”.
•
“dose addition and response addition ... represent default approaches for
toxicologically similar and toxicologically independent chemicals, respectively”.
Guidance section titled “Criteria for Dose Addition [DA] vs. Response Addition [IA]”
•
“The key criterion for choosing [DA or IA] is the similarity or independence among
the chemicals in the mixture”.
Thus, in the EPA Guidance the overall distinction is between similarity and independence, for
use of DA and IA respectively. The concept of dissimilarity is not emphasised.
WHO/IPCS: framework for the risk assessment of combined exposure to multiple
chemicals (Meek et al. 2011)
The WHO/IPCS framework is described in more detail in section 14.3. The framework
presents dose additivity as the default assumption for estimating risk in all tiers (Meek et al.
2011) and notes that “The use of dose additivity is considered conservative based on analysis
of empirical results for effects of combined exposure including to chemicals that induce
critical effects by different modes of action (US EPA, 2007; EFSA, 2008; European
Commission, 2010).” The use of independent action is not considered in any tier of this
framework, consequently no guidance is given, or needed, to address the question of similar
and dissimilar chemicals.
However in higher tiers, use of MOA information is proposed but not qualified by
consideration of the use of independent action to cumulate refined groups. Tier 2 allows for
the refinement of the assessment group “through consideration of increasingly more specific
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
information on mode of action or other factors on which to base the grouping (e.g., molecular
modeling). For some substances (e.g., cholinesterase inhibitors), this may be at the level of a
molecular target.” or the use of relative potency factors based on a selected index chemical.
Tier 3 can incorporate “increasingly refined information on mode of action, including both
kinetic and dynamic aspects. These can include both physiologically based pharmacokinetic
(PBPK) and biologically based dose–response (BBDR) models”
If the refinement in Tier 2 or 3 results in one group being replaced by two or more subgroups,
it becomes possible for risk to be underestimated unless there is a means of aggregating the
risk of the subgroups. If the risk is aggregated by DA, then the use of subgrouping becomes
trivial since DA would be used within AND between the subgroups and would be
mathematically equivalent to the use of DA on the whole group. IA could be used between
subgroups, with DA used within the subgroup, if there are experimental or theoretical grounds
to expect independence of effects of each subgroup, but the methods for doing this within
CRA are not currently used or generally available.
This chance of underestimating risk would not occur when risk estimates are refined through
refined exposure assessments, consequently we consider that exposure assessments should be
as refined as possible before grouping on the basis of mode/mechanism information is
introduced. In this way the risk estimate may become acceptable before reliance on
contentious subgrouping becomes necessary.
14.1.
Commonalities of approaches
Several commonalities emerge from consideration of the various approaches to risk
assessment:
•
Use of a tiered approach to deal with data availability, and a recognition that data
availability is a potentially limiting factor. The use of tiered approaches is discussed in
detail in section 14.3.
•
The need for a default assumption to allow assessment to proceed in the absence of
detailed mechanistic data
•
Agreement on the choice of additivity concepts: DA and IA
14.2.
Practical implications for CRA of chemicals with diverse uses and properties
and present in foods
Practical mixture assessment approaches have generally been formulated for pesticides rather
than other chemical groups. The need to extend methods developed with pesticides in mind to
other groups of chemicals in order to examine many, or perhaps all, types of chemical within
one CRA could be affected by the difference between pesticides, which are relatively well
studied and have a clear regulatory approach, and other groups of chemicals that are less well
studied. Issues include assumptions about typical exposure levels that may be different for
pesticides than for, for example, food additives which are deliberately added to food and food
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
contaminants which arise from environmental contamination. Environmental contaminants
may be less responsive to regulation and harder to manage, and may exceed their regulatory
limits, if they exist, more often than pesticides. Their chemical properties are also likely to
differ from chemicals designed for a purpose and intended to have a low human hazard, for
example the persistent organic pollutants (POPs) compared to modern pesticides, with rapid
human metabolism. The data available for pesticides on e.g. mode of toxic action, although by
no means complete, is also likely to be more substantial than that for other chemical groups.
Finally, although pesticides constitute a diverse group of chemicals, the inclusion of nonpesticides in an assessment is likely to increase diversity on most metrics, for example
chemical properties, uses, human exposure and toxicological effects etc.
The inclusion of all relevant chemicals in a CRA is scientifically important, since a division
of chemicals according to usage (e.g. pesticide, food additive etc) is unlikely to mirror a
division in toxicological risk. Thus, the scientific credibility of a CRA will be increased by
the inclusion of all relevant chemicals (those to which human exposure occurs) irrespective of
regulatory divisions.
14.3.
Assessment frameworks and tiering
Several ways of dealing with mixtures in chemicals risk assessment have recently been
proposed and discussed (IPCS 2009;Kortenkamp and Hass 2009;NRC 2008). Depending on
the quality of the data and the level of detail available for mixtures risk assessment (data poor
or data rich), tiering methods are useful for exploring the problem to be assessed. When
appropriate, more sophisticated models and associated supporting data can be used.
As a representative example for the approaches discussed in the literature we describe the
framework analysis for cumulative risk assessment proposed by (IPCS 2009).
The IPCS framework analysis begins by considering the nature of the exposures in question
and assesses whether the key mixture components that make up combined exposures are
actually known. This first step of the analysis may already expose knowledge gaps that
preclude continuation of a framework analysis.
Next, IPCS proposes to analyse whether exposure to specific chemicals is likely in the setting
relevant to the assessment. In some cases, chemicals may not be released into the
environment, or degradation is known to be very rapid, thus minimising the chances of
exposure.
If exposure is judged to be likely, the analysis can proceed by considering whether there are
opportunities for co-exposure within a relevant timeframe, and which substances are likely to
occur together. If the chances of co-exposure are negligible, the analysis may be terminated.
In cases where combined exposures are anticipated to take place, the next logical question to
be answered concerns the identity of chemicals that should be considered together in
cumulative risk assessment. At this stage, there are several options for proceeding with the
analysis: Some chemicals may be subjected to cumulative risk assessment because they occur
together in relevant exposure scenarios. In other cases it may be appropriate to select only
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
those that affect a common endpoint. The analysis can be further narrowed by restriction to
chemicals that affect a common endpoint by a common mode of action.
To avoid that unnecessary resources are invested to reach decisions about the risk assessment
issue under investigation, the IPCS framework has adopted a hierarchical structure, with
several tiers, depending on data quality, data demand and level of detail that is accessible. At
the lower tiers, relatively crude assumptions are made about exposures and potencies of the
chemicals that make up the mixture to be assessed. If the lower tiers already indicate
negligible risks, no further efforts are investigated to refine the analysis, and the assessment is
terminated.
At the lowest tier, Tier 0, crude and semi-quantitative exposure estimates may be used for the
development of a HI. Accordingly, the ADIs entered into the calculation of a HI may not
necessarily be comparable, having been derived from a variety of different endpoints, or by
applying UFs in inconsistent ways. At this stage, it may also be assumed that all chemicals in
the mixture are as potent as the most potent chemical present, if more appropriate data are not
available. Although these simplifications and inconsistencies may not be in accord with the
assumptions of the DA concept, they reflect the realities of chemical safety testing which is
geared towards identifying the most sensitive toxicities of a chemical. As a result, potency
information in relation to the same adverse outcome, as required by DA, is often not available
and this may limit the analysis. The procedure is considered to be sufficiently conservative
and protective, given that the input values are based on critical effects that occurred at the
lowest dose for each chemical. In a risk characterisation step, the margin between estimated
exposure and hazard is considered as a decision basis for determining whether a more refined
analysis is required.
If the Tier 0 HI is judged to be too large, the analysis can proceed to Tier 1, with the aim of
introducing refinements wherever possible. At this tier, the assumptions on exposures may
still be deterministic and may reflect worst case assumptions. For hazard assessments, the
IPCS document judges it permissible to assume that chemicals have the same potency as the
most potent known chemical present in the mixture, should better data not be available.
Where possible, more accurate estimates of potency may be incorporated, such as benchmark
doses. Alternatively, PODIs may be calculated. The risk characterisation step determines
whether further refinements of analysis should be conducted.
In Tier 2 analyses, deterministic exposure assessments are further refined, by incorporation of
measured data, additional parameters and more realistic scenarios. Hazard assessments may
focus on more restrictive assumptions about chemicals to be included in the analysis, e.g. by
including more information about modes of action, or the nature of endpoints that are affected
by the mixture components. Only PODs in relation to common adverse outcomes may be
included in Tier 2, or an analysis based on RPFs, with an index chemical, may be attempted.
IPCS suggests adopting the PODI method at this stage, with a consideration of the margin
between exposure and hazard as a basis for decisions as to whether higher tier assessment are
required.
As an example for a Tier 3 analysis, IPCS proposes the use of probabilistic exposure
assessments. Tier 3 assessments for hazard incorporate increasingly refined information about
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
the mode of action of the chemicals in question. According to the IPCS proposal, Tier 3 risk
characterisation steps still only decide whether a refined analysis is required.
The stage where risk management measures, rather than further refinements of the analysis,
should be envisaged is left open in the IPCS document.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TASK5: PROPOSE AN APPROACH TO CRA (DISSIMILARITY)
15.
A tiered framework analysis for combinations of dissimilarly acting chemicals
In fulfillment of Task 5 of the project, the purpose of this section is to propose a sciencebased approach for performing cumulative risk assessments of chemicals in food that act
through dissimilar modes of action. The science base, and our analysis, of it was presented in
the preceding tasks.
The proposal is based on three elements:
•
•
•
Cumulative risk assessment methods derived from DA,
A tiered approach following the broad principles that underpin the IPCS framework
analysis (IPCS 2009), and
An orientation towards criteria for the grouping of dissimilarly acting chemicals that
are based on common adverse outcomes.
The application of cumulative risk assessment methods derived from DA to combinations
of chemicals with dissimilar modes of action is essentially driven by pragmatism. As shown
in chapter 13.4, the predictions for combination effects that can be made by using DA and IA
do not differ significantly in most cases encountered in practice, and the factors that drive
these prediction differences are well understood. DA also generally yields the more
conservative mixture effect predictions, and can be regarded as sufficiently protective in most
situations. Examples where IA was more conservative, and produced an accurate prediction,
could not be located in the scientific literature (see section 13.1.5). Furthermore, risk
assessment methods based on IA are currently not in use and are yet to be developed (see
section 13.6.2). This, together with the fact that the data requirements for applying IA are
hard to meet in practice, argues for an extension of DA based cumulative risk assessment
methods also to mixtures composed of chemicals with dissimilar modes of action.
Tiered approaches to cumulative risk assessment can avoid unnecessary expenditure of
resources by offering the possibility of discontinuing the analysis on the basis of crude and
simple assumptions about exposures and hazards when margins of safety (or exposure) are
judged to be sufficient. In this way, lengthy, but largely unproductive efforts of refining the
analysis can be avoided.
It is clear that mixture toxicology cannot aggregate disparate effects. It is impossible to define
the joint effect of e.g. a liver toxicant and a pulmonary toxicant, if both these toxicities do not
find expression in a common adverse outcome, even though the adverse outcome may arise
through very different mechanisms. For this reason, cumulative risk assessments for
dissimilarly acting combinations of chemicals should be based on common adverse
outcomes as the organising principle of grouping for the purpose of defining common
assessment groups. At lower tiers of the analysis, chemicals with ADIs derived from disparate
effect endpoints may be considered together, as suggested by IPCS (2009). But as the tiers
become more refined, the mixture components should be grouped according to common
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
adverse outcomes, as much as possible. This may lead to the exclusion of some substances as
the analysis moves to the next higher tier.
15.1.
A unified approach to cumulative risk assessment for similarly and dissimilarly
acting chemicals
Our proposed application of cumulative risk assessment methods derived from DA also to
combinations of dissimilarly acting chemicals opens up the possibility of adopting a unified
approach to analysing combinations of chemicals, irrespective of their mode of action,
especially at lower tiers of the analysis. This can successfully overcome difficulties that may
prevent continuation of the analysis in situations where uncertainties about the modes of
action of chemicals block further efforts.
15.2.
Principal assumptions, simplifications and requirements
In common with the cumulative risk assessment approaches applied in regulatory practice
(EPA 2000), and the IPCS framework analysis (IPCS 2009), our proposal is based on a
number of assumptions which we make explicit, as follows:
1. The possibility of synergisms or antagonisms is disregarded. This assumption is
the direct consequence of the fact that the degree of synergism or antagonism cannot
be predicted quantitatively on the basis of the toxicity of the mixture components. All
mixture effect prediction methods and accordingly, all cumulative risk assessment
methods, assume additivity. Considering that the likelihood of synergisms is relatively
small (Boobis et al. 2011;EC 2009), the disregard for toxic interactions may be
regarded as sufficiently protective.
2. Simultaneous exposure to multiple chemicals is assumed. In numerous settings
encountered by humans there is simultaneous exposure to multiple chemicals. For
example, there is consumption of single food items that contain multiple chemicals
and, even when food items are consumed sequentially, the subsequent exposure of
body tissues to the chemicals contained within the items may be simultaneous. Strictly
sequential exposures are also a reality, but the risk assessment methods available for
cumulative risk assessment are not applicable to sequential exposure to multiple
chemicals. In theory and concept, methods for sequential exposures have yet to be
developed.
3. Exposures from non-food sources are not taken into account. It is estimated that
exposure via food accounts for the majority of chemical exposures currently
experienced by humans. However, exposures from non-food sources, including air,
consumer items, personal care products and pharmaceuticals also play a role. In view
of EFSA’s mission, the analysis can be restricted to food items as the source of
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
4.
5.
6.
7.
exposure, but may be extended to other exposure routes and sources, if deemed
necessary.
Potency estimates for mixture components are considered together at low tier
analyses, although they may be derived from different endpoints. Application of
DA (and IA) requires the use of potency estimates for the same adverse outcome as
input values. However, such input values are often not available because chemical
safety testing is geared towards identifying critical toxic effects. In practice, this
means that toxicity information for chemicals that occur together in mixtures often
derives from disparate endpoints. To enable assessments of cumulative risks, the
demand for potency estimates for the same endpoints is therefore relaxed, especially
for analyses at lower tiers. This simplification is in line with the principles of the
framework analysis suggested by (IPCS 2009).
It is assumed that the potency estimates entered into cumulative risk assessment
methods (e.g. ADIs, POD) describe doses associated with the same effect
magnitude. As discussed in section 13.2, the equations for DA are based on single
chemical effect doses for identical effect magnitudes. When applied to the PODs that
enter the mathematical expressions used in many cumulative risk assessment methods
such as HI or PODI, this means that all PODs should describe effect doses for the
same effect levels. In practice however, this demand cannot always be met, except in
the case of benchmark doses which are defined in relation to the same effect levels.
The effects associated with NOAELs that form the basis for ADIs by combination
with UFs are normally not known. To make cumulative risk assessment methods
workable despite these knowledge gaps, ADIs and PODs are taken as if they described
effect doses for the same effect magnitude.
Potency estimates can be derived from different tests, performed under different
conditions. In the interest of consistency, the evaluation of experimental mixture
effects by using the concepts of DA or IA should utilise effect data for all the mixture
components that were gathered under the same experimental conditions, with the same
animal strains. If this condition is not fulfilled, a bias may be introduced into the
analysis, leading to erroneous determinations of mixture effects in terms of additivity,
synergy or antagonism. Cumulative risk assessment however has to rely on data that
were produced in the context of single chemical testing, under widely varying
experimental conditions, even when the same strains were used, so that the demand of
consistency of data cannot be realised in practice. To allow continuation of cumulative
risk assessments, this demand is therefore relaxed.
Data on exposures and potency must be recorded by using the same dose metric.
The same dose metric (e.g. intake and potency as mg/kg d) must be used to allow
utilization of the formula for HI or PODI.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.3.
Rules for step-wise refinements as the analysis moves to the next higher tier
Cumulative risk assessment is based on at least three components: (1) selection of a defined
number of chemicals, on the basis of criteria derived from the exposure setting under
investigation or other grouping criteria, (2) data about exposures of the chemicals in the
mixture to be assessed, and (3) estimates about their potency. Should application of a
cumulative risk assessment approach indicate risks that are deemed to be unacceptable, the
analysis is refined in a step-wise fashion. As a general rule, it is proposed that only one of the
above three components is refined at a time, as the analysis moves to the next higher tier. The
most practical component should be refined first, for example the component that will require
the fewest pragmatic assumptions.
15.4.
Elements of cumulative risk assessment in a tiered framework analysis
In common with single chemical risk assessment, there are three elements of cumulative risk
assessment: Exposure assessment, hazard assessment and risk characterization.
Exposure assessment compiles exposure information about all the chemicals considered
together. Depending on the refinement of the analysis, this can be based on quite crude
assessments at lower tiers or on probabilistic data at higher tiers.
Hazard assessment is based on potency estimates of the chemicals in question. According to
the level of detail possible, this will rely on ADIs, or on PODs for specific endpoints,
depending on the aim of the analysis and the grouping criteria for the common assessment
group.
The risk characterization step ends with a decision about continuation of the analysis, with
further refinement. The decision criterion is whether the HI exceeds 1, or whether the margin
between hazard and exposure is judged to be sufficient. At some stage of the analysis the risk
characterization step should yield a decision criterion for risk reduction measures (see below).
15.5.
Framework analysis: initial considerations
As suggested in the IPCS framework (IPCS 2009), the analysis begins with considerations of
the nature of the exposures in question and assesses whether the key mixture components that
make up combined exposures are actually known. This first step may already expose
knowledge gaps that preclude continuation of the analysis.Risk assessment approaches that
start from known exposures may have limitations for use in the setting of regulatory limits,
which may need to be set without knowledge of exposures. In particular, although the
regulatory limits for single agents may be set by ‘reversing’ the risk assessment procedure,
the same is not true for the regulation of mixtures. Approaches to this issue have been
discussed elsewhere (EFSA 2009c).
Continuation of the framework analysis should be driven by the likelihood with which coexposures to chemicals can occur. Accordingly, the analysis should proceed by considering
which chemicals may occur together in the setting under investigation, initially without
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
consideration of their toxicological profile. It is conceivable that the analysis reveals that the
complete array of chemicals of concern present in e.g. a food item is not known. In such
cases, the analysis may proceed on the basis of known chemicals, but the uncertainty
introduced by knowledge gaps may have an impact on the magnitude of margins of exposure
that are deemed to be acceptable.
15.5.1. Tier 0
At the lowest tier, Tier 0, all chemicals that occur together in the exposure setting under
investigation are considered, irrespective of the effects they elicit.
Commensurate with the (low) quality of data that find entry in the analysis at this stage, it is
suggested that a hazard index (HI) should be constructed.
Crude and semi-quantitative exposure estimates may be used for the development of a HI.
Similarly, the potency estimates entered into the calculation of a HI may also be quite basic.
For example, in line with the IPCS (2009) proposal, it may be assumed that all chemicals in
the mixture are as potent as the most potent constituent, if more appropriate data are lacking.
Thresholds of Toxicological Concern (TTC) may also be used at this stage, with the aim of
bridging data gaps. If available, ADIs may be entered into the calculation of a HI, but there is
no need to enter ADIs consistently for all chemicals considered in Tier 0. The ADI values
may be derived from a variety of different endpoints and species, and may include different
UFs.
During the risk characterisation step, the margin between estimated exposure and hazard is
considered as the decision basis for determining whether a more refined analysis is required.
If the HI exceeds 1, the analysis should proceed to Tier 1.
15.5.2. Tier 1
In agreement with the proposed IPCS framework analysis, all chemicals relevant to the
exposure scenario under investigation are considered in Tier 1, irrespective of the effects they
produce, and without consideration of the modes of action involved. No chemical that is a
component part of the exposure scenario under investigation is excluded from the analysis at
this stage.
The assumptions on exposures may still be deterministic and may reflect worst case
assumptions, but they should rely on measured values as much as possible.
For hazard assessments, estimates of potency for each chemical are incorporated, such as
ADIs or benchmark doses. Simplifying assumptions, such as TTC or potencies similar to the
most toxic known substance present in the mixture should be abandoned. The potency
estimates may be for a variety of different endpoints, and can be derived from studies with a
variety of different test species. Alternatively, and if possible, hazard assessments may be
based on PODs for all the chemicals considered, with application of the PODI in the risk
characterisation step (for further details see below).
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The risk characterisation step determines whether further refinements of analysis should be
conducted. This should be pursued if the HI (or PODI) is larger than 1, or if the margin of
safety is judged to be inappropriate.
15.5.3. Tier 2
In the preceding tiers of the analysis, no chemical present in the mixture under consideration
was excluded from the analysis. In Tier 2, this restriction may be relaxed, by considering for
the first time the effect profile of chemicals, with the intention of creating assessment groups
of chemicals. Since this step of the analysis is by no means trivial, and may be hampered
severely by data gaps, we propose to deal with this topic initially by excluding those
chemicals that are known not to produce a chosen common adverse outcome (see case study
3, section15.8.3). Consequently, the assessment group may include chemicals where there is a
degree of uncertainty as to whether they can contribute to a common effect. This is to avoid a
situation in which the analysis lacks conservatism by defining too narrow common
assessment groups on the basis of positive effect criteria.
As before, the exposure assessment should rely on measured data. The hazard assessment
may utilise ADIs (or, alternatively PODs) that were derived for common endpoints,
irrespective of any consideration of mode of action.
If the risk estimates exceed acceptable levels, the analysis may be refined and proceed to Tier
3.
15.5.4. Tier 3
At this stage, the analysis may adopt more restrictive criteria about common adverse
outcomes, and may define groupings of chemicals for assessment on the basis of
phenomenological effect criteria. As is commensurate with an analysis for dissimilarly acting
chemicals, these groupings will still be made irrespective of any modes of action or
mechanisms that may underlie the induction of the effects of interest.
The exposure assessment element may utilize probabilistic data, if available.
Tier 3 assessments for hazard may incorporate increasingly refined potency estimates for the
specific endpoints that form the basis for defining groups of chemicals for assessment. At this
stage, PODs may be used, with the aim of constructing a PODI. In the interest of consistency
of analysis, the PODs should derive from the same animal species. In this way, the analysis
approaches a level of detail similar to that exercised during the evaluation of experimental
mixture studies, where all single chemicals and the mixture were studied under similar
conditions. Nevertheless, the PODs may still reflect some differences in terms of data quality
and experimental standards.
In Tier 3, construction of a PODI, rather than a HI, may be regarded as more appropriate, for
the following reasons:
It is assumed that the PODs that form the basis of the analysis are of similar quality. In this
case, it is appropriate that the aggregation for mixture effects should be conducted at the level
of experimental data, by calculating a PODI. This practice achieves a high level of
Supporting publications 2012:EN-232
147
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
consistency that comes nearer to the application of DA as a mixture assessment concept in
experimental mixture studies. It realises a high level of transparency, by avoiding the
introduction of too many assumptions (e.g. use of different uncertainty factors in inconsistent
ways).
In the risk characterisation stage of the analysis, an uncertainty factor can be used for the
entire group of chemicals, or, alternatively, a margin of exposure can be determined.
15.6.
When should the risk characterisation step result in risk management
measures?
As described in section 14.3, the IPCS framework analysis does not define the stage where
the risk characterisation step should result in proposing risk management measures, rather
than further refinements of the analysis.
In our proposed framework analysis for combinations of dissimilarly acting chemicals, the
only option for introducing further refinements of the grouping of chemicals beyond Tier 3 is
in considering the modes of action of the mixture components under consideration. However,
such refinements are called for when the analysis is for combinations of similarly acting
chemicals, and would not be appropriate for dissimilarly acting chemicals. For this reason, it
is proposed to discontinue the analysis at Tier 3, with a recommendation for risk management
measures if the margin of exposure or the size of the PODI is deemed unacceptable. By
default, that should be the case with margins of exposure smaller than 100, or with a PODI
larger than 1 (if constructed by using the default UF of 100).
However, in view of the data gaps that might preclude proceeding to Tier 3, it is a point of
serious consideration whether risk management measures should also be envisaged at the end
of Tier 2. This may be justified in cases where further refinements of analysis are not possible
in the foreseeable future, because existing data gaps are unlikely to be filled.
15.7.
Criteria for the grouping of chemicals in CRA at higher tiers
Key features of our proposed unified approach are that
•
•
all chemicals relevant to the exposure scenario under investigation are considered
together at lower tiers of the analysis, irrespective of their presumed mode of action,
and that
the grouping of chemicals according to their ability to affect common adverse
endpoints, with exclusion of substances judged to be irrelevant, only takes place at
higher tiers.
This raises the question as to how such groupings should be conducted, and whether the data
necessary to achieve well-founded groupings are available.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Here, we approach this issue by first considering debates and proposals about groupings for
similarly acting mixtures in the context of tiered approaches to CRA. Several case studies
have been published where the applicability of the IPCS framework analysis was investigated.
These studies have considered chemicals with quite similar structural features, e.g.
polybrominated biphenyl ethers or carbamates (IPCS 2009). EFSA has published a document
about grouping criteria for triazoles (EFSA 2009c).
It has been noted that cumulative risk assessment based on quite narrow criteria of chemical
similarity may underestimate risks by excluding other substances from the analysis that might
nevertheless contribute to cumulative effects, even by differing modes of action (EC
2009;NRC 2008). There is experimental evidence that the concept of DA produces reliable
estimations of combination effects with mixtures composed of chemicals with diverse modes
of action (Christiansen et al. 2009;Crofton et al. 2005;Rider et al. 2010).
For this reason, broader grouping criteria, based on similarity of effect in a phenomenological
sense, have been proposed for similarly acting combinations, e.g. for the assessment of antiandrogenic substances (NRC 2008). Such grouping criteria can also be utilised for
dissimilarly acting chemicals, e.g. from Tier 2 onwards. However, the development of
phenomenological grouping criteria based on common adverse outcomes has to be done with
due consideration of physiological processes.
As was discussed in the NRC report, phenomenological grouping criteria should be based on
a sound understanding of the physiological processes that lead to a common adverse outcome.
In the case of combinations of antiandrogenic substances, the decisive factor is disruption of
androgen action in fetal life, which can occur by different mechanisms and modes of action.
Blocking of the androgen receptor, suppression of androgen synthesis and disruption of
steroid-metabolising enzyme systems are processes of relevance, which lend themselves as
criteria for the grouping of chemicals that should be considered together for an assessment of
cumulative anti-androgenic effects. Accordingly, substances shown to
•
•
•
•
antagonise the androgen receptor,
suppress androgen synthesis,
interfere with steroid-converting enzymes, and
suppress androgen receptor expression
should be considered as candidates for a common assessment group for chemicals affecting
male sexual differentiation. The identification of candidate chemicals can be based on both in
vitro and/or in vivo data, and suitable assays are available. While these are relatively clear
criteria for the identification of candidate chemicals that should be subjected to CRA, the
implementation of CRA in practice has to rely on the provision of further data about
exposures and potency estimates. Both these elements currently present bottlenecks, as
discussed by (Kortenkamp and Faust 2010). Exposure estimates for many chemicals
identified as in vitro androgen receptor antagonists are rudimentary, or not available at all.
The input data for the hazard assessment step of CRA should be derived from in vivo studies
where the ability of candidate chemicals to affect the above anti-androgenic endpoints has
been assessed. Such data are currently available only for a very limited set of chemicals.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
A comparable level of understanding is currently achieved perhaps with thyroid-disrupting
chemicals, but the bottlenecks that prevent meaningful application of CRA are similar.
Thyroid hormone levels are determined through a complex interplay between several factors,
including uptake of iodine from dietary sources which is required for the synthesis of the
hormone, transport of iodine to the thyroid gland, synthesis of the precursor of active thyroid
hormone, transport of the precursor to tissues and deiodinisation of the precursor to the active
thyroid hormone. Suppression of thyroid hormone levels by disruption of any of these
processes is regarded as the decisive factor. There is experimental evidence that chemicals
capable of interfering with these processes through multiple mechanisms elicit combined
effects well predicted by DA (Crofton et al. 2005;Crofton 2008). Accordingly, elements of
these physiological steps can serve as criteria for the inclusion of chemicals in a common
assessment group. Thus, substances shown to
•
•
•
•
Interfere with iodine uptake,
Suppress synthesis of the precursor hormone,
Block deiodinisation of the precursor, or
Antagonize hormone action at the receptor level
are candidates for inclusion in a common assessment group, and suitable in vitro and in vivo
assays for the identification of such chemicals are available. However, as with antiandrogenic chemicals, the implementation of CRA is hampered by a lack of data for the
critical input values, i.e. exposure estimates and potency estimates for the above thyroidrelevant endpoints.
The physiological processes that lead to other common effects are comparatively poorly
understood. As a way of dealing with these knowledge gaps in the context of cumulative risk
assessment, discussions are underway to group chemicals together according to the effects
they exert on target organs, or on systems, such as the reproductive system. Such grouping
strategies have to be applied judiciously and with great care. For example, all carcinogens,
irrespective of the tissues in which they induce tumours, could be considered together. While
such a grouping is sensible at lower tiers of the analysis, a better understanding of the
physiological processes involved will have to drive the grouping at higher tiers. Similarly, all
substances known to produce reproductive and developmental toxicity could be considered
together at lower tiers. However, reproductive toxicity encompasses a broad range of different
effects, ranging from reductions in fertility to malformations of organs. At higher tiers, such
diverse effects will have to be differentiated.
It is beyond the scope of this project to define detailed phenomenological grouping criteria.
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.
Case studies
To illustrate various aspects of the proposed approach to cumulative risk assessment we
provide three case studies in this section. Each study is intended to illustrate the approach and
should not be considered as an attempted cumulative risk assessment.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.1.
Case study 1: pesticides and contaminants in lettuce
In this case study, we explore the application of CRA to the analysis of specific food items.
In a monitoring program in the year 2007 the German Federal Agency for Risk Assessment
(BfR) investigated pesticide residues and environmental contaminants such as heavy metals
and nitrate in various food items including salad. Butterhead lettuce was analysed for
potential residues of 510 different pesticides. Most of the samples were from Germany.
However, some samples came from other European countries. A total of 63 samples were
taken. There are no published data on specific substances and their levels that can be
attributed to a single sample. Arithmetic mean levels (MW) of pesticides and contaminants in
lettuce are documented and it is indicated, in how many samples (absolute/ percentage of all)
the levels were below, at, or above the analytical detection limit.
For an assessment of potential health effects due to the residues and contaminants, we
compared the levels in lettuce with the substance specific reference values (ADI for pesticides
and TDI for contaminants). ADI-values were mostly reported by EFSA (EFSA 2011b).
Where necessary, those were supplemented by earlier listings from BfR (BfR 2007). For lead
the former “provisionally tolerable weekly intake” (PTWI) of 25 µg/kg x w was used (3.5
µg/kg x d), not accounting for the uncertainties as documented by EFSA (EFSA 2010b).
Similarly, for cadmium the “tolerable weekly intake” (TWI) of 2.5 µg/kg x d was used and
transformed into a daily dose of 0.4 µg/kg x d (EFSA 2009b). For copper there is an
“acceptable range of oral intake” (AROI) with the upper limit of “more than 2-3 mg per adult
and day” (WHO 1998), which was accounted for by a reference value of 5 mg/d x 60 kg =
0.08 mg/kg x d. For nitrate, an ADI was available (EFSA 2008a)
Table 14 provides a list of the residues and contaminants in lettuce, the number of samples,
where those substances were found above detection limit, the content as arithmetic means
(MW) and the respective reference values (ADI).
We used these survey results to analyse the data for the impact of mixture effects as assessed
according to the tiered approach proposed in this report (section 15).
Three assessment scenarios (A, B, C) were assumed alternatively:
Assessment
scenario
Exposure characterization
Scenario A
Isolated analysis of lettuce consumption; no other sources for identified
“Lettuce only” residues and contaminants in food considered; reference point: ADI
Analysis of lettuce consumption assuming that also other sources for all
“Lettuce plus substances within the mixture contribute to intake; EFSAs data on pesticide
uptake via different food commodities (expressed as % of ADI and
food
calculated by using mean pesticide concentrations and mean intake values
background”
for each commodity) were used as background values; reference point:
flexible (background dependent) fraction of ADI
Scenario B
Scenario C
Analysis of lettuce consumption assuming that only a fixed minor fraction
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
“Lettuce
allocated
fraction”
of the ADIs for all mixture components is allocated to this food commodity
as permissible maximum intake; reference point: fixed fraction of ADI
(quote of ADI)
For all scenarios we assumed that the average consumption of salad in Germany is 23 grams
per day (EFSA 2008a).
15.8.1.1.Assessment Scenario A
In this scenario the analysis focuses exclusively on lettuce as the sole source of intake
(butterhead lettuce; “lettuce only”). Other food items are not included in the analysis. Tier 1
of the proposed framework analysis is applied. The results are shown in Table 15.
10 pesticides or nitrate or metal contaminants were found with a HQ of ≥ 0.1% (shaded in
Table 15), with the assumption of a mean consumption of lettuce 23 grams/person/day and a
body weight of 60 kg). The maximum number of pesticides identified in one and the same
sample was 13, but the identity of the substances was not reported in the original report (BVL
2008a). Under the (conservative) assumption that all of the 53 different substances are present
in a single sample, and using the arithmetic mean of the concentrations [mg/kg] found from
all 63 samples for each of the substances, a HI of 28% can be calculated. The main
contribution is from nitrate, with a HQ of 22.8%. About 2% of the HI may be attributed to
pesticide residues.
Some of the substances were not analysed in all of the 63 samples. The data for pirimicarb
were not included as only 10 samples were analysed for this pesticide. As is appropriate for a
low tier analysis, there was no grouping of the pesticides and contaminants in terms of
common adverse outcomes. It may be assumed that the number of contaminants and
pesticides present in one and the same lettuce sample is well below the maximum number of
13 found in one sample, and this will reduce the hazard index accordingly. However, certain
substances may have been present in single samples with concentrations well above the
arithmetic mean, on which the above estimate was based. In any case, the analysis
demonstrates that most of the HI is due to a single substance exposure (nitrate). All other
analysed pesticides and contaminants contribute only to a minor degree to the HI. Therefore,
in this case no higher tier analysis is necessary. However, it may be argued that scenario A is
insufficiently conservative, and that the risk characterization step leads to a false negative
decision as it does not cover pesticides’ exposure due to other food items.
15.8.1.2.Assessment Scenario B
To deal with the shortcomings of the previous scenario A, it was now assumed that the
pesticides and contaminants found in lettuce are also taken up via other food items (“lettuce
plus food background”). To account for uptake via other food sources (for details see next
paragraph), the ADIs (TDIs) for each substance were corrected accordingly. The HQ were
calculated by using lower ADIs, corrected for the intake from other food items (ADI x (1calculated exposure via other food items in %). As the amount of each residue and
contaminant in other food items differed, substance specific allocations were calculated. For
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
example, the ADI for dimethoate was lowered by 14.8% because total uptake of dimethoate
via different food commodities was 14.8% of the ADI as calculated by EFSA (EFSA 2011b)
and accordingly, the allocated ADI therefore was 85.2% of the original ADI. Similarly, 72%
auf the TDI for cadmium was attributed to other sources with a remaining allocated TDI of
28% of the original TDI. Table 16 shows the results of this calculation.
The data on the fraction of ADI assigned to exposure to the respective pesticides via other
food commodities was taken from “The 2009 European Union Report on Pesticide Residues
in Food”(EFSA 2011b). EFSA has performed an analysis on the total uptake of pesticide
residues via food based on monitoring data from the Member States and consumption data
from the PRIMo database. These data were transformed into a figure representing a fraction
of the respective ADI. We used the data for GEMS/Food Cluster E diet for further analysis.
Note that this fractional ADI includes exposure to all food items, also including lettuce.
Therefore, there is some double counting: the fraction of the ADI available for salad exposure
depends on the fraction for all “other” food items, which again includes salad. Within this
analysis it was not possible to adjust for this imprecision. However, the bias introduced by
this omission is negligible.
For lead, an average uptake via food of 0.34-1.24 µg/kg x w is reported (EFSA 2010a)
corresponding to up to 35% of the PTWI. Cadmium exposure via food may be very high and
may already reach or exceed the TWI (EFSA 2009b). A fraction of 72% as a minimum
assumption was assigned to other food items but salad. For copper, 1-2 mg/adult intake is via
drinking water and food (WHO 1998), which corresponds to about 40% of the AROI reported
above. For nitrate, a high percentage of the ADI is taken up via drinking water and cured meat
(close to 20%). More than 66% of the ADI has to be assigned to fruits and vegetables, in this
case after exclusion of salad (calculated from (EFSA 2008a)). From this analysis we assigned
a fraction of 86.4% of the ADI to other food items.
Calculation of the HI with the modified ADIs or TDIs gave a value of 180%, clearly
exceeding 100%. The majority of the HI can be attributed to nitrate (167%), 10% to cadmium
and the pesticide residues only make up 2% of the HI.
For scenario B pesticide uptakes via other commodities were taken from the 2009 European
Union report on pesticide residues in food (EFSA 2011b). The individual allocations of the
ADI of each pesticide with respect to GEMS/Food cluster diet E was regarded in this
calculation. Again, nitrate was the most relevant single substance in this scenario B as it was
already in scenario A. Nitrate uptake via other fruits and vegetables besides lettuce and the
uptake via drinking water and cured meat accounts for 86.4% of ADI and only 13.6% of the
ADI remain for lettuce in scenario B. Based on the mean concentration of nitrate in lettuce
and a mean consumption of 23 g/d the respective hazard quotient for nitrate (167.4%) clearly
exceeded 100%. Because the high hazard index is driven by one single substance, no higher
tier analysis was conducted. Increases of residues or contaminants in other food will
simultaneously decrease the available fraction for lettuce and would therefore change the
hazard index.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.1.3.Assessment Scenario C
This analysis is based on lettuce consumption assuming that only a fixed minor fraction of the
ADIs for all mixture components is allocated to this food commodity as permissible
maximum intake. This fixed fraction of ADI is used as reference point. For the calculations in
Scenario C we assumed that 5% of the ADI of each individual pesticide or contaminant is
allocated for the uptake via lettuce. This percentage is based on the average lettuce
consumption (23 g) in relation to consumption of vegetables and fruits, which may contain
residues of pesticides (about 400g/d). Table 17 provides the results of this assessment
including the hazard ratios.
The hazard index with 5% of the respective ADIs as reference points clearly exceeds 100%
and reaches a value of 559% or 5.59. However, as in scenarios A and B, only one substance
(nitrate) contributes to most of this load. After excluding nitrate and the metal contaminants a
HI of only 40% can be calculated, based on the pesticides alone.
Because of the disproportionate influence of nitrate the result would not change significantly
if the analysis proceeded to a higher tier assessment. The lettuce scenario indicates that there
is an issue with one single, or perhaps two substances, nitrate and cadmium. The procedure
adopted in this scenario has been used for drinking water, where the WHO proposes 10% of
the ADI for the respective substances as a reference point.
15.8.1.4.Higher Tier Assessments
In this case study we have presented variations of a Tier 1 assessment for lettuce as food item.
Despite the fact that there are only small incentives to proceed to higher tier analyses, we
would like to sketch out some elements that might be useful for Tier 2 analyses. In Table 18
we illustrate a first grouping step. To this end, we selected randomly a small subset of 5
pesticides, with the aim of exploring methodological aspects of the approach.
We grouped the pesticides and contaminants according to common adverse outcomes by
considering potential target organs of the substances within the mixture. As an example for
the proposed procedure we analysed the dossiers for the 5 chosen pesticides (PRAPeR, DAR,
EFSA substance specific scientific opinions) with the aim of deriving further information
about common adverse outcomes and their points of departure. The points of departure and
the corresponding endpoints that formed the basis for the estimation of an ADI were listed,
and the ADI then taken for the calculation of HI (see Table 18). For toxicities that were not
critical for the estimation of ADIs, reference doses (RfD) were estimated as follows:
Bifenthrin is a clear neurotoxicant (tremors in dogs and rats at elevated doses) and the
NOAEL for this effect was used to establish the ADI. Bifenthrin also shows carcinogenic
effects in animals at much higher doses by a nongenotoxic mode of action and is classified as
a low potency suspected carcinogen (classified Canc. Cat. 2, CLP). The carcinogenicity is
observed at doses more than 10 times higher than those associated with neurotoxicity.
Accordingly, a RfD for carcinogenicity was assumed to be 10 times higher than the ADI,
derived by multiplying the ADI with a factor of 10. These factors are listed in Table 18 for
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
other toxicities. If, for a specific organ, no toxicity was detected, the multiplier was set to
zero.
As is apparent from Table 18, there are 3 neurotoxicants and 3 agents leading to sensitisation
among the selected 5 pesticides. At higher tiers of the analysis, these target organ effects
would have to be considered in more detail. For nephrotoxicity, glomerular and tubular
nephrotoxic substances should be distinguished if this distinction is clearly supported by the
data.
15.8.1.5.Conclusions
The main findings of this case study can be summarized as follows:
The number of substances occurring together in one food item and therefore contributing to a
mixture effect is often limited (in this case the maximum number of pesticides in one lettuce
sample was 13).However, some substances might have been missed, due to inappropriate
analytical detection limits. On the basis of more sensitive detection limits, the number of
substances to be considered in CRA would increase accordingly, but the relevance of this in
terms of CRA is unclear, because only relatively small HQ would result, with limited impact
on the HI.
A few single substances (such as nitrate or cadmium) with widespread environmental
occurrence may constitute the majority of a HI and are of health concern on their own,
independent of any combination effects. Reducing exposures to these substances will have a
strong impact on the overall risk estimate.
A higher tier approach in mixture risk assessment is not warranted, if one single substance
contributes disproportionately to the HI.
We demonstrated the feasibility in principle of a tiered approach by considering a small
subset of substances of this case study. In fact, PRAPeR- and DAR-documents provide useful
quantitative information to be used in higher tiers mixture risk assessment. However, we did
not pursue such an analysis here, because the HI could be explained in terms of a few single
chemicals. Nevertheless, some detailed guidance should be established in the future on how to
derive RfD for specific common adverse outcomes different from those that form the basis for
estimating ADIs.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 14: Residues and contaminants in butterhead lettuce (BVL 2008a;BVL 2008b)
% of samples
above detect
limit
n > detect.
Limit
Content
Arithmetic mean
MW [mg/kg]
EFSA 2011 ADI *)
[mg/kg bw x d]
Acetamiprid
2
4.17
0.00265
0.07
Azoxystrobin
1
1.59
0.00067
0.2
Bifenthrin
4
6.35
0.00271
0.015
Boscalid (Nicobifen)
20
41.67
0.27896
0.04
Methyl bromide
47
74.60
6.03575
1
Folpet
2
3.17
0.00938
0.1
Carbendazim
1
1.59
0.00032
0.02
Chlorpyrifos-methyl
2
3.17
0.00041
0.01
Chlorpyrifos
2
3.17
0.00016
0.01
Clothianidin
2
8.00
0.00156
0.02
Cypermethrin [ISO]
1
1.59
0.00008
0.015
12
19.05
0.01425
0.03
Deltamethrin
3
4.76
0.00173
0.01
Diethofencarb
1
1.59
0.00019
0.43
Dimethoate
2
3.17
0.00056
0.0003
Dimethomorph
3
4.76
0.00048
0.05
24
38.10
0.27632
0.05
Endosulfan (-A, -B, sulfate)
1
1.59
0.00003
0.006
Fenhexamide
1
1.59
0.0119
0.2
Fludioxonil
7
11.11
0.01852
0.37
Indoxacarb
2
3.17
0.00008
0.006
Imidacloprid
2
4.17
0.00027
0.06
35
55.56
0.60378
0.06
1
2.08
0.00004
0.015
11
17.46
0.01865
0.005
Metalaxyl-M
7
11.11
0.00663
0.08
Linuron
1
2.08
0.00002
0.003
Methomyl
2
4.17
0.00619
0.0025
Myclobutanil
1
1.59
0.00005
0.025
Oxadixyl
1
1.59
0.00016
0.05
Cyprodinil
Thiram**)
Iprodione
Iprovalicarb
Cyhalothrin, l-
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Pendimethalin
3
4.76
0.00078
0.125
Pirimicarb
3
4.76
0.00051
0.035
Procymidone
13
20.63
0.01335
0.0028
Propamocarb
18
37.50
0.86969
0.29
Propyzamide
7
11.11
0.00138
0.085
Pymetrozine
2
4.17
0.00067
0.03
11
22.92
0.07048
0.03
Pyrimethanil
1
1.59
0.00365
0.17
Tebufenpyrad
7
11.11
0.00408
0.01
Terbuthylazine
1
1.82
0.00022
0.002
Thiamethoxam
4
8.33
0.01148
0.026
Tolylfluanid
7
11.11
0.02421
0.1
Tolclofos-Methyl
8
12.70
0.02935
0.064
Trifluralin
1
1.59
0.00021
0.015
Vinclozolin
2
3.17
0.00087
0.005
Pencycuron
2
5.26
0.00353
0.0200
Spinosad
1
2.08
0.01838
0.0240
Spinosyn A
1
3.70
0.02767
0.0200
Spinosyn D
1
3.70
0.005
0.0200
Lead
5
33.30
0.008
0.0036
Cadmium
15
100.00
0.02633
0.0004
Copper***)
14
93.30
0.61
0.0800
Nitrate
59
93.65
2197
3.7000
Pyraclostrobin
*) for some substances EFSA-ADIs were not available. In those cases other ADI or TDI reported in the
literature were used. See text for further references.
**) Only “Dithiocarbamate” listed in report - Thiram used as Dithiocarbamate with lowest ADI from those
permitted for use in Germany
***) further contaminants (Thallium, Zinc) were excluded because of low concentrations and no adequate
reference values (TDI) available
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 15: “Lettuce only” assessment scenario (tier 0). Hazard Index with no influence of
exposure to other food items
MW
[mg/kg]
ADI
Hazard quotient % (MWx23g/d) /
(mg/kg bw) (ADI*60kg)
Acetamiprid
0.00265
0.07
0.00145
Azoxystrobin
0.00067
0.2
0.00013
Bifenthrin
0.00271
0.015
0.00693
Boscalid (Nicobifen)
0.27896
0.04
0.26734
Methyl bromide
6.03575
1
0.23137
Folpet
0.00938
0.1
0.00360
Carbendazim
0.00032
0.02
0.00061
Chlorpyrifos-methyl
0.00041
0.01
0.00157
Chlorpyrifos
0.00016
0.01
0.00061
Clothianidin
0.00156
0.02
0.00299
Cypermethrin [ISO]
0.00008
0.015
0.00020
Cyprodinil
0.01425
0.03
0.01821
Deltamethrin
0.00173
0.01
0.00663
Diethofencarb
0.00019
0.43
0.00002
Dimethoate
0.00056
0.0003
0.07156
Dimethomorph
0.00048
0.05
0.00037
Thiram
0.27632
0.05
0.21185
Endosulfan (-A, -B, sulfate)
0.00003
0.006
0.00019
0.0119
0.2
0.00228
Fludioxonil
0.01852
0.37
0.00192
Indoxacarb
0.00008
0.006
0.00051
Imidacloprid
0.00027
0.06
0.00017
Iprodione
0.60378
0.06
0.38575
Iprovalicarb
0.00004
0.015
0.00010
Cyhalothrin, l-
0.01865
0.005
0.14298
Metalaxyl-M
0.00663
0.08
0.00318
Linuron
0.00002
0.003
0.00026
Methomyl
0.00619
0.0025
0.09491
Fenhexamide
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Myclobutanil
0.00005
0.025
0.00008
Oxadixyl
0.00016
0.05
0.00012
Pendimethalin
0.00078
0.125
0.00024
Pirimicarb
0.00051
0.035
0.00056
Procymidone
0.01335
0.0028
0.18277
Propamocarb
0.86969
0.29
0.11496
Propyzamide
0.00138
0.085
0.00062
Pymetrozine
0.00067
0.03
0.00086
Pyraclostrobin
0.07048
0.03
0.09006
Pyrimethanil
0.00365
0.17
0.00082
Tebufenpyrad
0.00408
0.01
0.01564
Terbuthylazine
0.00022
0.002
0.00422
Thiamethoxam
0.01148
0.026
0.01693
Tolylfluanid
0.02421
0.1
0.00928
Tolclofos-Methyl
0.02935
0.064
0.01758
Trifluralin
0.00021
0.015
0.00054
Vinclozolin
0.00087
0.005
0.00667
Pencycuron
0.00353
0.0200
0.00677
Spinosad
0.01838
0.0240
0.02936
Spinosyn A
0.02767
0.0200
0.05303
Spinosyn D
0.005
0.0200
0.00958
Lead
0.008
0.0036
0.08519
0.02633
0.0004
2.80366
Copper
0.61
0.0800
0.29229
Nitrate
2197
3.7000
22.76171
HI
27.96%
Cadmium
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 16: Assessment Scenario B: “Lettuce plus food background”. Calculation of
Hazard Index (tier 0)
MW
[mg/kg]
Exposure
via all food
commodities
analysed
(%ADI,
WHO Food
Cluster E)*
ADI
(mg/kg
bw)
HQ %
(MWx23g/d
)/
(ADI*60kg) HQ % based on adjusted ADI
Acetamiprid
0.00265
0.07
0.10
0.00145
0.0015
Azoxystrobin
0.00067
0.2
0.10
0.00013
0.0001
Bifenthrin
0.00271
0.015
1.20
0.00693
0.0070
Boscalid
(Nicobifen)
0.27896
0.04
0.50
0.26734
0.2687
Methyl bromide
6.03575
1
0.00
0.23137
0.2314
Folpet
0.00938
0.1
0.20
0.00360
0.0036
Carbendazim
0.00032
0.02
0.50
0.00061
0.0006
Chlorpyrifos-methyl
0.00041
0.01
1.40
0.00157
0.0016
Chlorpyrifos
0.00016
0.01
2.20
0.00061
0.0006
Clothianidin
0.00156
0.02
0.00
0.00299
0.0030
Cypermethrin [ISO]
0.00008
0.015
1.20
0.00020
0.0002
Cyprodinil
0.01425
0.03
0.40
0.01821
0.0183
Deltamethrin
0.00173
0.01
2.50
0.00663
0.0068
Diethofencarb
0.00019
0.43
0.00
0.00002
0.0000
Dimethoate
0.00056
0.0003
14.80
0.07156
0.0840
Dimethomorph
0.00048
0.05
0.20
0.00037
0.0004
Thiram
0.27632
0.05
3.50
0.21185
0.2195
Endosulfan (-A, -B,
sulfate)
0.00003
0.006
1.90
0.00019
0.0002
0.0119
0.2
0.10
0.00228
0.0023
Fludioxonil
0.01852
0.37
0.00
0.00192
0.0019
Indoxacarb
0.00008
0.006
0.90
0.00051
0.0005
Fenhexamide
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a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Imidacloprid
0.00027
0.06
0.20
0.00017
0.0002
Iprodione
0.60378
0.06
0.50
0.38575
0.3877
Iprovalicarb
0.00004
0.015
0.10
0.00010
0.0001
Cyhalothrin, l-
0.01865
0.005
2.10
0.14298
0.1461
Metalaxyl-M
0.00663
0.08
0.10
0.00318
0.0032
Linuron
0.00002
0.003
0.40
0.00026
0.0003
Methomyl
0.00619
0.0025
1.20
0.09491
0.0961
Myclobutanil
0.00005
0.025
0.30
0.00008
0.0001
Oxadixyl
0.00016
0.05
0.00
0.00012
0.0001
Pendimethalin
0.00078
0.125
0.00
0.00024
0.0002
Pirimicarb
0.00051
0.035
0.20
0.00056
0.0006
Procymidone
0.01335
0.0028
1.80
0.18277
0.1861
Propamocarb
0.86969
0.29
0.00
0.11496
0.1150
Propyzamide
0.00138
0.085
0.00
0.00062
0.0006
Pymetrozine
0.00067
0.03
0.00
0.00086
0.0009
Pyraclostrobin
0.07048
0.03
0.00
0.09006
0.0901
Pyrimethanil
0.00365
0.17
0.10
0.00082
0.0008
Tebufenpyrad
0.00408
0.01
0.40
0.01564
0.0157
Terbuthylazine
0.00022
0.002
0.00
0.00422
0.0042
Thiamethoxam
0.01148
0.026
0.00
0.01693
0.0169
Tolylfluanid
0.02421
0.1
0.10
0.00928
0.0093
Tolclofos-Methyl
0.02935
0.064
0.20
0.01758
0.0176
Trifluralin
0.00021
0.015
0.00
0.00054
0.0005
Vinclozolin
0.00087
0.005
0.90
0.00667
0.0067
Pencycuron
0.00353
0.0200
0.00
0.00677
0.0068
Spinosad
0.01838
0.0240
0.00
0.02936
0.0294
Spinosyn A
0.02767
0.0200
0.00
0.05303
0.0530
Spinosyn D
0.005
0.0200
0.00
0.00958
0.0096
Lead
0.008
0.0036
35.00
0.08519
0.1311
0.02633
0.0004
72.00
2.80366
10.0131
Copper
0.61
0.0800
40.00
0.29229
0.4872
Nitrate
2197
3.7000
86.40
22.76171
167.3655
HI
27.96%
180.05%
Cadmium
* Data from (EFSA 2011b)
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 17: Assessment Scenario C (fixed fraction”). Calculation of hazard index (tier 0)
MW
[mg/kg]
ADI
(mg/kg bw) HQ % (MWx23g/d) / (0,05*ADI*60kg)
Acetamiprid
0.00265
0.07
0.02902
Azoxystrobin
0.00067
0.2
0.00257
Bifenthrin
0.00271
0.015
0.13851
Boscalid (Nicobifen)
0.27896
0.04
5.34673
Methyl bromide
6.03575
1
4.62741
Folpet
0.00938
0.1
0.07191
Carbendazim
0.00032
0.02
0.01227
Chlorpyrifos-methyl
0.00041
0.01
0.03143
Chlorpyrifos
0.00016
0.01
0.01227
Clothianidin
0.00156
0.02
0.05980
Cypermethrin [ISO]
0.00008
0.015
0.00409
Cyprodinil
0.01425
0.03
0.36417
Deltamethrin
0.00173
0.01
0.13263
Diethofencarb
0.00019
0.43
0.00034
Dimethoate
0.00056
0.0003
1.43111
Dimethomorph
0.00048
0.05
0.00736
Thiram
0.27632
0.05
4.23691
Endosulfan (-A, -B, sulfate)
0.00003
0.006
0.00383
0.0119
0.2
0.04562
Fludioxonil
0.01852
0.37
0.03837
Indoxacarb
0.00008
0.006
0.01022
Imidacloprid
0.00027
0.06
0.00345
Iprodione
0.60378
0.06
7.71497
Iprovalicarb
0.00004
0.015
0.00204
Cyhalothrin, l-
0.01865
0.005
2.85967
Metalaxyl-M
0.00663
0.08
0.06354
Linuron
0.00002
0.003
0.00511
Methomyl
0.00619
0.0025
1.89827
Myclobutanil
0.00005
0.025
0.00153
Oxadixyl
0.00016
0.05
0.00245
Fenhexamide
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Pendimethalin
0.00078
0.125
0.00478
Pirimicarb
0.00051
0.035
0.01117
Procymidone
0.01335
0.0028
3.65536
Propamocarb
0.86969
0.29
2.29918
Propyzamide
0.00138
0.085
0.01245
Pymetrozine
0.00067
0.03
0.01712
Pyraclostrobin
0.07048
0.03
1.80116
Pyrimethanil
0.00365
0.17
0.01646
Tebufenpyrad
0.00408
0.01
0.31280
Terbuthylazine
0.00022
0.002
0.08433
Thiamethoxam
0.01148
0.026
0.33851
Tolylfluanid
0.02421
0.1
0.18561
Tolclofos-Methyl
0.02935
0.064
0.35159
Trifluralin
0.00021
0.015
0.01073
Vinclozolin
0.00087
0.005
0.13340
Pencycuron
0.00353
0.0200
0.13532
Spinosad
0.01838
0.0240
0.58714
Spinosyn A
0.02767
0.0200
1.06068
Spinosyn D
0.005
0.0200
0.19167
Lead
0.008
0.0036
1.70370
0.02633
0.0004
56.07315
Copper
0.61
0.0800
5.84583
Nitrate
2197
3.7000
455.23423
HI
559.22%
Cadmium
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded following
a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety
Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the
issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Bifenthrin
82657-04-3
Fenhexamid
126833-17-8
Fludioxonil
131341-86-1
Dimethoate
60-51-5
Cypermethrin
52315-07-8
TIER 2
0
0
0
0
0
0
TIER 3
1
0
0
1
1
3
1
0
0
1
1
3
0
0
1
0
0
1
4
10
1
0
0
3
0
0
0
0
0
0
4
1
0
1
0
3
0
0
0
0
2
1
2
5
1
2
1
5
ADI [mg/kg x d] Cancer
Reproductive
Developmental
Hematotox
Bones
Nephrotox
Hepatotox
Neurotox
Sensitisation
Respiratory
CAS No
Substance
Name
Table 18: Higher tier mixture risk assessment – example: subset of pesticide data
10
0
0
0
0
0.015 0.2 0.37 0.001 0.04 1
n
of
endpoint
specific
relevant
substances
Calculate hazard ratio and hazard index only for those substances, for which the value in the respective column is > 0. Use
original ADIs as reference points (e.g. for kidney effects three substances are regarded as relevant). Highest hazard index
characterises the mixture effect
Calculate hazard quotients and hazard index only for those substances, for which the value in the respective column is > 0
(relevant endpoint; see TIER 2). Use ADI x value given in the respective matrix field above as your reference point (e.g., use 4
x ADI(Bifenthrin) = 4 x 0.015 = 0.6 for kidney effects of Bifenthrin)
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Authority and the author(s), awarded following a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety Authority is subject. It may not be
considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.2. Case study 2: Mycotoxins in food commodities
Mycotoxins are toxic secondary metabolites produced from several species of fungi. They are
widely distributed food and feed contaminants which pose a serious problem in food and feed
safety. Several mycotoxins may occur simultaneously in food and feed which results in a
combined intake of mycotoxins by humans and animals. Maximum levels for mycotoxins in
food have only been derived for single mycotoxins or mycotoxins belonging to the same
group like aflatoxins B1, B2, G1, G2, fumonisins B1 and B2 or T2-toxin and HT-2 toxins
(see http://ec.europa.eu/food/food/chemicalsafety/contaminants/index_en.htm). Until now,
risk assessment focuses on single substances or closely related mycotoxins such as T2-toxin
and its metabolite HT-2 toxin. Whether the combined intake of numerous mycotoxins from
structurally similar (e.g. trichothecenes) or different groups may possibly result in an
increased risk to human health shall be investigated in the following case study.
While the previous case study on pesticides and contaminants present in lettuce focused on
one single food item, this case study will investigate the cumulative exposure to several
mycotoxins from different commodities.
In the past several scientific cooperation (SCOOP) tasks have focused on the collection of
data about the levels of mycotoxins in food and on the assessment of dietary intake of these
substances. For this case study the SCOOP data on fusarium toxins, patulin and ochratoxin A
(EC 2002a;EC 2002b;EC 2003) have been evaluated. Data reported from the United
Kingdom (UK) on the dietary mycotoxin concentrations, food consumption and dietary
intake have been selected for this case study due to the comprehensive measurements and
detailed analysis of consumption data for several groups of the population.
Food commodities were analysed for the following trichothecene mycotoxins:
Group A trichothecenes
T-2 toxin
HT-2 toxin
T2-triol
Diacetoxyscirpenol
Neosolaniol
Monoacetoxyscirpenol
Verrucarol
Group B trichothecenes
Deoxynivalenol
Nivalenol
3-Acetyldeoxynivalenol
15-Acetyldeoxynivalenol
Fusarenon-X
Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin and HT-2 toxin are the most relevant
substances from this group. They are included in the further analysis because they are the
only members from this group for which reference values exist (see below).
Additionally the fusarium toxins zearalenone and fumonisin B1 and B2 as well as ochratoxin
A and patulin were considered for cumulative risk assessment.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The tolerable daily intakes (TDIs) as derived by the Scientific Committee on Food (SCF), the
Joint FAO/WHO Expert Committee on Food Additives (JECFA) or EFSA are reported in
Table 19.
In Table 20 the total dietary intake of DON, NIV, T-2 toxin, HT-2 toxin, zearalenone, patulin
and OTA for adults and toddlers as calculated by UK in the SCOOP reports are summarized
(EC 2002a;EC 2002b;EC 2003). All food commodities monitored for mycotoxins were used
for this calculation (for details see SCOOP reports). Total dietary intake was calculated using
consumption data of the whole population differentiated by age groups and not only of
consumers. For this example the adults (16-64 years) and toddlers (1.5-4.5 years) were
selected, the UK analysis was much more differentiated. Further, mean values for mycotoxin
concentrations in food commodities have been used. The following principles have been
applied for the calculation of the mean mycotoxin concentrations in food:
•
•
•
All data provided were used.
If limit of detection (LOD) and limit of quantification (LOQ) were available, LOD/2
were used for results lower than the LOD. For results between LOD and LOQ,
numerical values, if available, were used.
If only LOQ was available, or if numerical values between LOD and LOQ were not
available, LOQ/6 was used for values below the LOQ.
Here, we provide the results of a Tier 1 analysis for mycotoxins. In this analysis, TDIs
derived from a variety of endpoints, in a variety of species were used (see Table 19). The
total dietary intake values were then compared with the TDIs for each mycotoxin to calculate
HQ for the individual mycotoxins. The HQs were then summed up to the hazard indices (HI)
for each age group regarding all mycotoxins. Whereas the HQs were all < 1 for the individual
mycotoxins, a HI of 0.84 or 1.01 was calculated for females or males, respectively. Total
intake data for fumonisin B1 and B2 are missing in this table, because the UK did not report
these data to SCOOP. But it was stated in the SCOOP report that fumonisin intake can reach
up to 14% of the TDI in adults (regarding the results from the other countries), which would
result in HIs exceeding 1.
Even without taking account of fumonisin, a HI of 2.5 was calculated for toddlers. Had it
been possible to consider fumonisin, the HI would have been even larger, because fumonisin
intake may reach 22% of the TDI according to SCOOP. The trichothecenes DON, T-2 toxin
and HT-2 toxin contribute strongly to the HI.
There is therefore the need to refine the analysis further by proceeding to Tier 2. In Tier 2, it
would be necessary to group mycotoxins according to common effects or common target
organs. As shown in Table 21, all selected mycotoxins cause reproductive and immunotoxic
effects. Additional endpoints affected by these mycotoxins are e.g. growth retardation,
haematotoxicity, neuro-, renal- and liver toxicity. These are the most relevant endpoints
affected by the mycotoxins but do not include all toxicological effects elicited by these
substance. For further information see (EFSA 2006;SCF 2000a;SCF 2000b;SCF 2002;WHO
1995). Regarding the two apical endpoints reproductive toxicity and immunotoxicity it
becomes obvious that none of these mycotoxins can per se be excluded from the cumulative
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
risk assessment. Calculation of a HI by using ADI with respect to immunotoxic or
reproductive toxic effects in Tier 2 would end up with the identical result as obtained in Tier
1.
A further refinement of this CRA would be desirable, e.g. by using NOAELs or BMDs for
immune- or reproductive toxicity. For a better comparison POD for the same reproductive or
immunotoxic effects should be selected. As these values are not easily available from the
toxicity reviews without further evaluation of the scientific literature we stopped the
elaboration of this example at this point.
This example demonstrates that the proposed CRA method can also be applied to situations
where cumulative exposure from several commodities takes place. It may be a useful tool to
investigate possible risks which might be lost in single substance evaluations.
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 19: Tolerable daily intakes for mycotoxins (source: (EC 2002a;EC 2002b;EC
2003)
TDI (µg/kg
Key study
bw)
Deoxynivalenol a
Most sensitive endpoint
Point of
departure
Extrapolation
factor
1 µg/kg bw
2-year feeding study in
mice
reduced growth
0.1 mg/kg
bw/day
(NOAEL)
100
Long term dietary
study in mice
reduced growth and
haematotoxic effects
0.7 mg/kg
bw/day
(LOAEL)
1000
haematotoxic effects
0.03 mg/kg
bw/day
(LOAEL)
500
hormonal effects
40 µg/kg bw
(NOEL)
200
Nivalenol a
0.7 µg/kg bw
T-2 toxin/HT-2 toxin a
0.06 µg/kg bw
3-week feeding study
with T-2 toxin in swine
Zearalenone a
0.2 µg/kg bw c
15-day study in pigs
Fumonisin B1+B2 a
2 µg/kg bw
chronic
Effects in rats and equine
0.2 mg FB1/kg
100
toxicity/carcinogenicit
leukoencephalomalacia
bw/day
y in rats
(NOAEL)
b
Patulin
0.4 µg/kg bw d
combined reproductive Not specified in SCOOP
43 µg/kg bw/day 100
toxicity, long term
report
(NOEL)
toxicity/carcinogenicit
y study in rats
Ochratoxin A e
90-day feeding study
Nephrotoxicity
8 µg/kg bw/day
450
0.017 µg/kg
in female pigs
(LOAEL)
bw/day e
Notes. a: evaluation by the SCF; b: evaluation by JECFA; c: temporary TDI; d: PMTDI: provisional maximum
TDI; e: evaluation by EFSA (EFSA 2006); f: calculated on basis of a tolerable weekly intake (TWI) of 0.120
µg/kg bw/week
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 20:
Mycotoxin uptake in the population of the United Kingdom (further
explanations in the text) and cumulative risk assessment
Age Group
Average male adult
(16-64 years)
Average female
adult (16-64 years)
Average toddler
(1.5-4.5 years)
Mycotoxin
Total dietary intake
(ng/kg bw/day)
TDI
(ng/kg bw/day)
HQ (Intake/TDI)
DON
NIV
T-2 + HT-2 toxin
Zearalenone
Patulin
OTA
276
40
33
14.06
11
0.53
1000
700
60
200
400
17
0.276
0.057
0.55
0.0703
0.0275
0.0312
HI: 1.012
DON
NIV
T-2 + HT-2 toxin
Zearalenone
Patulin
OTA
195
27
29
12.98
11
0.53
1000
700
60
200
400
17
0.195
0.0386
0.4833
0.0649
0.0275
0.0312
HI: 0.840
DON
NIV
T-2 + HT-2 toxin
Zearalenone
Patulin
OTA
796
119
54
53.85
120
1.42
1000
700
60
200
400
17
0.796
0.17
0.9
0.269
0.3
0.0835
HI: 2.52
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
x
x
x
x
X
X
X
x
(x)
x
x
x
x
x
x
Supporting publications 2012:EN-232
Liver toxicity
Haematotoxicity
x
x
x
x
Renal toxicity
Growth
retardation
x
x
x
x
Neurotoxicity
Immunotoxicity
DON
NIV
T-2 toxin
HT-2
toxin
ZEA
FB1+2
PAT
OTA
Reproductive
toxicity
Table 21: Most relevant apical toxicological effects affected by the selected mycotoxins
x
x
x
x
x
x
x
171
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.3. Case study 3: dietary exposure to pesticides
15.8.3.1.Description of dataset
For this case study we illustrate the stages of a cumulative risk assessment using the exposure
and risk data provided for 34 pesticides in JMPR reports from 2009 and 2010 (JMPR
2009;JMPR
2010),
available
online
http://www.fao.org/agriculture/crops/core‐
themes/theme/pests/pm/jmpr/jmpr‐rep/en/. Rather than providing realistic risk estimates
for exposure scenarios involving pesticides, the aim of this case study is to explore whether,
and to what degree, lack of data might hamper continuation of the analysis. Accordingly, the
chemicals included in this case study were selected on practical grounds, that data about them
were available in either of the two most recent JMPR reports. This does not indicate that
chemicals that were not included in this analysis should not be considered, nor that the
included chemicals are considered priorities for assessment. The guiding approach used is the
hazard index (HI). Annex 3 of each JMPR report contains international estimated daily intakes (IEDI) which are
expressed as the percentage of the respective ADI, for the 13 cluster diets in the Global
Environment Monitoring System-Food contamination and assessment programme
(GEMS/Food, http://www.who.int/foodsafety/chem/gems/en/index1.html). These numbers
are equivalent to the hazard quotients (HQ) required for the calculation of a hazard index
(HI), see section 13.6.1.1, but depicted on a percentage scale so that the range from 0 to 1 is
expressed as 0 to 100%. The compiled data is presented in Table 22 and has been rescaled so
that the HQ and HI are on a unitary scale, not percentage, in which values greater than 1
indicate a greater than acceptable risk for the single chemical (HQ) or mixture (HI).
15.8.3.2.Tier 1: HI analysis (all ADIs)
A hazard index (HI) analysis of the full dataset is presented in Table 23. In line with a low
tier analysis, all available ADIs, irrespective of the endpoints they are based on, are
considered together.
In the main body of the table cells highlighted in red indicate when a single substance has a
HQ in excess of 1, indicating a greater than acceptable risk for that chemical alone. HQ
values exceeded one in two cases (1.1, 1.4), both for chlorpyrifos methyl. The JMPR note
that values above 1 (or 100, when expressed at percentages) should not necessarily be
interpreted as giving risk to a health concern because of the conservative assumptions used in
the assessments (JMPR 2009). However, for CRA it is important to note that if any single
chemical has a HQ of >1 then the HI will exceed 1 irrespective of the contribution of any
other chemicals.
The final row of the table shows the HI, the sum of all individual HQs. This row is coloured
according to the extent by which the HI exceeds 1. The table shows that the HI exceeds 1 for
all thirteen GEMS diets, and ranged from 1.3 to 5.2. In no case was the HI solely driven by a
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
single chemical exceeding a HQ of 1. In low tiers, this analysis would indicate the need for
further refinement and precludes the conclusion of acceptable risk.
Further approaches to interpreting the HI are shown in the remainder of the case study, which
use the data subset for GEMS diet E, which includes Austria, Belgium, Croatia, Czech
Republic, Denmark, France, Germany, Hungary, Ireland, Luxembourg, Malta, Netherlands,
Poland, Slovakia, Slovenia, Switzerland and United Kingdom of Great Britain and Northern
Ireland.
15.8.3.3.Interpretation of HI and HQ values
Table 23 shows the ADI, exposure (JMPR IEDI) and HQ of each chemical in the GEMS diet
E. In this case the HI, the sum of all HQs, is 2.9, indicating that an acceptable level of risk
cannot be concluded and that refinement of the assessment is required.
This data is shown graphically in Figure 19. The graph shows the distribution of exposure
(IEDI, panel A) and risk (ADI, panel B) values that produce the HQ distribution (panel C and
panel D, log scale). The graphs are ordered along the x-axis according to decreasing HQ
values, and thus the cumulative HQ graph (panel E) shows the build up of contributions
towards the HI of 2.88. The HI is indicated by a solid vertical line.
The cumulative HQ graph (panel E) can be used to explore the amount of refinement that
would be required to enable the HI to attain values below 1 (indicating acceptable risk). The
graph provides access to the two extreme approaches to the refinement, in the best case the
chemicals that contribute most to the estimate would be refined first, and the dotted line at
1.88 indicates the chemicals (all those with bars below and the first bar to exceed the value of
1.88) that would need to be refined for the HI to drop below 1. In this case, the first seven
chemicals would need to be refined and if their individual HQs still contributed significantly
after refinement then additional chemicals would need refinement. Conversely in the worst,
most inefficient, case, if refinement was directed at those chemicals not making a significant
contribution (those on the right of the graphs) then the dotted line at 1 indicates the chemicals
(all those chemicals with bars crossing the dotted line) that would need to be refined for the
HI to be below 1. In this worst case, 32 chemicals out of 34 would need to be refined,
indicating that if the first two chemicals in the graphs, haloxyfop and chlorothalonil, are not
refined then the HI is very likely to exceed 1.
Consideration of the HQ spectrum can reveal where refinement could have the most effect
and is likely to be more efficient than refining all chemicals or taking a random approach.
When the spectrum is skewed (as can be seen in panel C) then focused refinement is even
more likely to be more efficient than other approaches, since a few chemicals are
disproportionately contributing to the risk estimate, and should be targeted by refinement. An
understanding of the extent to which refinement might be effective could be used to decide if
refinement of this approach is practical (for example how many chemicals must undergo risk
refinement) or if an exposure refinement might have more impact.
Supporting publications 2012:EN-232
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.3.4.HI analysis using TTC values (pseudo tier 0 analysis)
The preceding HI analysis was possible because all of the included chemicals have been
assigned ADIs. If a chemical has not been assigned an ADI it has been suggested that the
TTC approach (see section 13.6.3) could be applied. We have therefore applied the TTC
approach to the data set used in this case study to explore its impact. We note that the TTC
concept is not intended for use when chemicals have been assigned ADIs, as is true here;
however we are using the approach here to assess its impact rather than to perform an
assessment. We have also used the extreme case when TTC values are used for all of the
chemicals in the assessment.
SMILES codes for each pesticide were retrieved from the PPDB (pesticide properties
database, http://sitem.herts.ac.uk/aeru/footprint/index2.htm) and ToxTree software (version
2.5.0, Ideaconsult Ltd.; http://sourceforge.net/projects/toxtree/) was used to assign chemicals
to the appropriate Cramer class based on their SMILES code. All of the chemicals were
assigned to Cramer class III, which receives a TTC value of 90 ug/person per day. To
perform a HI assessment using TTC values, abbreviated to HI (TTC), HQs are calculated by
dividing IEDI values by TTC values rather than ADIs, and are termed here HQ (TTC). This
analysis is presented in Table 23 which compares the HI values calculated using ADIs
(previous section) or TTC values (this section).
Impact of TTC values on the risk estimate
The HI for GEMS diet E was 2.9 when calculated using the ADI, use of the TTC values
results in a HI (TTC) of 37.8. The TTC might typically be applied to only a few components
in a mixture assessment, and the impact of the use of TTC values should be assessed
whenever they are used. This is because risk estimates driven by the use of TTC values are
clear candidates for refinement before action is taken. In this case study, the mean HQ (TTC)
was 1.1, indicating that if a TTC value was applied to a single chemical, with the remainder
assessed using their ADIs, then on average, the HI would be expected to exceed one in all
cases. HQ (TTC) values actually ranged from 0.001 to 5.6 and the impact of smaller values
would not greatly affect an overall HI, whilst larger values could be fully responsible for a
risk estimate exceeding 1. The distribution of these values would only apply generally if
other chemicals follow the same risk (ADI) and exposure distribution of the pesticides used
in this case study.
Use of the conservative TTC approach to bridge data gaps in higher tiers may be impractical
when the exposure level of a mixture component is close to the TTC and it may be that either
refinement of the exposure assessment or the acquisition of toxicological data is preferred
over the propagation of TTC values though to higher tiers.
The interpretation of HQ (TTC) values is less valuable than interpretation of HQ (ADI)
values because the TTC does not represent a measure of risk in the same way that the ADI
does. Consequently HI (TTC) and HQ (TTC) are a measure of exposure with an
approximation of risk based solely on chemical structure. HQ and HI values derived from
ADI or TTC are also not proportional to each other, see Figure 20. Therefore HQ (TTC)
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
values should not be strictly used for targeting refinement since it is probably the TTC values
themselves that require refinement, i.e., they would ideally be replaced by ADIs once a
suitable toxicity data set was available.
15.8.3.5.PODI (NOAELs)
The use of a point of departure index (PODI) approach is shown in Table 24. Table 24 lists
the NOAEL value for each chemical together with supporting information, including
uncertainty factor (UF) values. Ideally a PODI approach would use values for the point of
departure (POD) for the same endpoint, however in this example we have applied the
approach irrespective of the endpoint for illustration purposes.
For this case study PODI = 0.0305 (Table 24). Interpretation of a PODI requires
multiplication by a group uncertainty factor (UFG), which for this case study might be 100,
since it was the UF applied to 33 out of 34 of the pesticides. However the assignment of a
group uncertainty factor may require expert judgement and further deliberation rather than a
crude ‘averaging’ approach, as used in this case study for pragmatic reasons. Using an
uncertainty factor of 100, gives a PODI x UFG value of 3.05, which exceeds 1 and would be
considered to indicate a greater than acceptable risk.
The PODI x UFG value of 3.05 is greater than the value of the HI (2.88) because one
pesticide, triazephos, had a single chemical UF of 10 (because human toxicology data was
available), consequently the use of an UF of 100 in the PODI approach increases the
contribution of triazephos. This effect illustrates the inability to apply chemical specific UFs
in the PODI approach, which might be considered a limitation. However some commentators
consider that the ability to remove policy- driven UFs is in fact an advantage, allowing
subjective uncertainty considerations to be dealt with by the selection and application of a
UFG, and separating these calculations from calculations dealing with data-based potency
values, PODs (Wilkinson et al. 2000).
Interpretation of a PODI value can also be done through determination of a combined margin
of exposure (MOET). For this case study the MOET is 32.78 (the reciprocal of the PODI
value), well below a value of 100 or more which might be required for an acceptable MOET
in this situation, considering that the UF applied to 33 out of 34 of the pesticides was 100.
In low tiers, when data may not be available on specific endpoints there may be little
advantage in using a PODI approach. In cases when the chemicals being considered have the
same UF, the HI, PODI and MOET approaches should produce numerically identical results
(Wilkinson et al. 2000); and in cases when the UF differs there may need to be an expert
judgment as to the appropriate group UF to apply.
Supporting publications 2012:EN-232
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
15.8.3.6.Tier 2 (consideration of effects)
In the previous tier, all HQs were summed despite the fact that the ADI on which the HQ is
based derive from different endpoints and not all of the chemicals cause every possible toxic
effect. It seems logical that the next level of refinement would be to break down the HI so as
to only include chemicals that affect a common adverse outcome, but irrespective of their
mode of action. The implicit assumption made at this stage is that lack of consideration of
mode of action will capture those chemicals that exert their effects through a variety of
mechanisms.
However, realisation of this level of detail is less trivial than it initially seems and has
substantial data requirements that may be virtually impossible to satisfy. This is now
illustrated by reference to the dataset considered in this case study.
The data in Table 24 shows that the NOAEL for each chemical was derived from a different
endpoint. Unless the NOAEL was assigned for the same endpoint then further refinement
will require substantially more data. If the same endpoint was used then the risk would be
calculated using the ADI, the most sensitive endpoints, and the risk from any other endpoint,
occurring by definition at higher doses, could be disregarded because the risk would always
be less than that for the most sensitive endpoint. When different effects lead to NOAELs, the
data for all endpoints becomes required as chemicals can only be excluded from further
calculations if they are shown not to cause an effect. This is because if, e.g. all 34 pesticides
in this case study had a different effect responsible for the NOAEL then a number of the
chemicals could still share a common toxic effect at a dose only slightly above the dose
producing the NOAEL. If this were the case for a sufficient number of chemicals then the
cumulative risk of that effect might be unacceptable but would have been undetected if the HI
was broken down using groups based on the effect leading to NOAEL.
For this case study the data that is required would be the highest dose tested without effect
for every endpoint of toxicological interest. This is a substantial data requirement, both in the
experimental work required and the effort involved in compilation of the data. It was not
feasible to perform this data collection for this case study, since the data, if available, are
present in free text or tables in reports with an often inconsistent structure and published in
pdf format. None of these features facilitate data extraction and compilation.
This case study shows that a database of toxicological endpoints without summarisation to
the most sensitive end point is required for meaningful cumulative risk assessments.
Failure of the CRA
Since compilation of the data was not feasible, the assessment of this case study would fail at
this stage, with low tiers having failed to conclude acceptable risk (because HI > 1) and with
this tier being unable to complete due to data requirements. The data requirement of this
stage is sufficiently high that many assessments could fail at this stage. It is a characteristic of
our approach that this analysis should not feature in a very early stage of a CRA, since this
would predispose to early failure in many cases. Positioning of this large data requirement
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
allows a CRA to conclude acceptable risk if that can be achieved in low tiers with
conservative assumptions. The CRA will only fail in higher tiers when more data are required
to conclude acceptable risk than are available.
In order to illustrate how the assessment would continue we have now used a compromise
data set based on human health classifications assigned to each of the chemicals in the
Pesticide Properties DataBase (PPDB; http://sitem.herts.ac.uk/aeru/footprint/index2.htm).
15.8.3.7.Breakdown of HI analysis using PPDB health issues
The PPDB includes nine human health issues, which are listed in Table 25. The issues are not
regulatory categories and may not be suitable for regulatory purposes but they are used here
to illustrate a grouping approach that may refine the analysis. For the purpose of this case
study it is assumed that the PPDB health issues cover all the effects of interest, and that they
are assigned reliably. The PPDB states whether each chemical is known to cause an effect,
known not to cause the effect or if the data is unclear or unavailable. Consequently, it is
possible to exclude chemicals from calculation of a HI if they are known not to cause an
effect, and this is the approach to the grouping of chemicals for CRA that drives the analysis
at this stage. It would not be conservative to only include the chemical that are known to
cause an effect, since in many cases there will many chemicals for which the data are not
available or is unclear. Consequently data from all three situations (‘known’, ‘unclear’ and
‘no data’) are included, but their relative contribution is calculated so it can be seen how
much of the revised HI is due to known risks or to the conservative assumption of risk unless
the absence of risk has been stated. This analysis is shown in Figure 21.
Figure 21 shows the HI calculated for each health issue. The height of each bar is 2.88, which
is the HI based on all ADIs (see above). Each bar is then broken into 4 portions. The top
(white) portion can be discounted because it shows the HQs that derived from chemicals that
are known (according to the PPDB classification) not to cause the health issue. The height of
each bar when this portion is discarded can be compared to the critical value of 1 because the
graph background is colour coded (blue below 1, red above 1). For ease of viewing, this
graph is also shown in Figure 22 with the white portions removed. Portions of each bar
coloured in red indicate the sum of HQs for chemicals that PPDB classified as known to
cause an effect, portions coloured orange indicate the sum of HQs for chemical that have an
uncertain data set and those coloured grey indicate the sum of HQs for chemicals which lack
data.
Detailed assessment of the toxicological relevance of these breakdowns may be inappropriate
since the data may not be suitable for risk assessment, however the general pictures is that
this analysis approach allows the reduction of risk estimates when chemicals can be excluded
(when known not to have an given effect). In this case however, most risk estimates remained
above the critical value of 1, and require further refinement because the risk cannot be
deemed acceptable. For acetylcholinesterase inhibition the data set contained many chemicals
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
that were known not to cause the effect, and consequently the HI for this issues was reduced
below 1 (0.9), indicating an acceptable risk.
Use of reference values for each effect in place of ADI: adjusted ADIs
If the data requirements for this tier were met, then ADIs could be 1) grouped by relevant
chemicals and 2) replaced by use of the actual values for the effect under consideration. This
was not possible for this case study using PPDB classification due to the nonnumeric nature
of the classification, however this is a feature of the summary nature of the PPDB, and may
not be a concern in reality, i.e. if there is knowledge about testing for all endpoints there is
also likely to be dose information too.
15.8.3.8.Tier 3 (group according to known or plausible toxicological independence)
For this case study detailed mechanism or mode of action information for all of the potential
toxic effects was considered unlikely to be available. It seems likely that risk estimates can
be refined in other ways before this massively data intensive step is reached.
In the event of such information being available, the methods used throughout this case study
and which are based on DA, could be supplemented with methods based on IA. This would
be a high tier assessment. Use of approaches based on IA should be done in a ‘mixed model’
approach, which requires development. In such an approach all chemicals causing a given
effect are first grouped according to similarity and the group risk is estimated with methods
based on DA, and then the risk of all groups is aggregated using methods based on IA.
The case for application of approaches based on pure IA rests on assumptions that are
considered highly unlikely to ever be met, for example:
•
That the toxic effect being considered is observed in an assay and that the observed
effect can be caused by multiple, toxicologically independent routes.
•
That all components cause a common effect but each through a toxicological
independent route. In the case of a scenario with 10 components, this would imply 10
independent routes to one toxic effect.
15.8.3.9.Conclusions
In low tiers the HI approach using TTC or ADI values produced a HI greater than one,
indicating a greater than acceptable risk. Further refinement was not possible due to high data
requirements although the approach that could be used was illustrated using PPDB health
issue classifications.
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The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The HI is a composite of hazard and exposure data; however the overall driver of the HI
value would appear to be the number of chemicals included. Opinions vary as to what the
realistic human scenario should be, and this constitutes an important knowledge gap.
The case study identifies several issues for consideration:
•
For cumulative risk assessment toxicological data need to be accessible and collated
across endpoints and across chemicals. The data for hazard and exposure should be in
comparable metrics and available in an open, standardised format.
Data
summarisation and censoring should be avoided.
•
The available risk assessment approaches have differing data demands, and there
should be a realistic assessment of whether these requirements will be met before an
approach is employed, otherwise the likely outcome is that the assessment cannot be
completed due to data requirements.
•
Assessment approaches with high data demands should be placed high in framework
approaches to give the best chance of completing risk assessments at lower tiers
before high tiers that are likely to fail on data requirements are reached.
•
Refinement of exposure should be considered, and possibly applied before more
complex risk assessment approaches are employed.
•
The data requirements for the selection of risk assessment approaches based on IA
limit these approaches to very high tiers.
Supporting publications 2012:EN-232
179
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Table 22: Hazard quotients (HQs) and hazard index (HI) for pesticides from JMPR 2009, 2010
Supporting publications 2012:EN-232
Africa Latin America Far East 0
0
0.015
0.51
0.143
0.003
0.012
0.257
0.002
0.014
0
0.003
0.009
0.004
0.005
0.003
0.476
0.863
0.24
0.024
0.105
0.644
0.023
0.291
0.01
0.078
0.018
0.088
0.003
0.002
0.18
1.113
0.324
0.015
0.028
0.306
0.018
0.083
0.004
0.018
0.013
0.037
0.002
0.001
0.117
0.397
0.212
0.013
0.058
0.591
0.013
0.088
0.002
0.034
0.011
0.042
0.002
0.001
0.131
0.295
0.17
0.014
0.049
0.527
0.01
0.134
0.001
0.039
0.012
0.046
0.001
0.001
0.08
0.205
0.178
0.01
0.08
0.447
0.008
0.271
0.001
0.057
0.011
0.03
0.002
0.001
0.079
0.277
0.29
0.006
0.086
0.259
0.057
0.003
0.047
0.006
0.018
0.001
0.001
0.06
1.436
0.098
0.006
0.027
0.644
0.071
0.004
0.019
0.012
0.021
0.001
0
0.026
0.93
0.066
0.003
0.026
0.25
0.022
0.001
0.011
0.005
0.006
0.001
0
0.017
0.233
0.075
0.002
0.019
0.206
0.019
0.001
0.007
0.005
0.005
0.001
0
0.059
0.308
0.182
0.004
0.013
0.813
0.05
0.002
0.013
0.01
0.022
0.001 0.003 0 0.001 0.07 0.151 0.308 0.477 0.27 0.156 0.006 0.012 0.108 0.087 0.386 0.655 0.063 0.193 0.002 0.007 0.049 0.049 0.005 0.013 0.019 0.045 Europe/Latin America Africa M Latin America L Far East K Europe J Europe I Europe/Middl
e East H Africa/Middle East G Africa/Europe
/Middle East 2,6‐DICHLOROBENZAMIDE 2009 BENALAXYL 2009 BUPROFEZIN 2009 CHLOPRYRIFOS METHYL 2009 CYPERMETHRIN 2009 FENBUCONAZOLE 2009 FLUOPICOLIDE 2009 HALOXYFOP 2009 HEXYTHIAZOX 2009 INDOXACARB 2009 METAFLUMIZONE 2009 METHOXYFENOZIDE 2009 PROTHIOCONAZOLE 2009 SPIRODICLOFEN 2009 Hazard quotient (IEDI / ADI) GEMS/Food consumption cluster diets A B C D E F Africa Chemical name: GEMS region JMPR year 180
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may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights
of the authors.
Combined actions of chemicals in food through dissimilar modes of action
ZOXAMIDE BIFENAZATE BIFENTHRIN BOSCALID CADUSAFOS CHLORANTRANILIPROLE CHLOROTHALONIL CLOTHIANIDIN CYPROCONAZOLE DICAMBA DIFENOCONAZOLE ETOXAZOLE FENPYROXIMATE FLUBENDIAMIDE FLUDIOXONIL FLUOPYRAM MEPTYLDINOCAP NOVALURON THIAMETHOXAM TRIEAZOPHOS HI(sum of HQs): 2009 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 2010 Supporting publications 2012:EN-232
0
0.025
0.082
0.113
0.006
0.001
0.2
0.006
0.005
0.008
0.008
0.001
0.005
0.03
0.019
0.012
0
0.069
0.006
0.26
1.828
0.003
0.226
0.2
0.402
0.003
0.004
0.4
0.015
0.016
0.003
0.106
0.006
0.06
0.245
0.018
0.065
0.001
0.459
0.036
0.023
5.159
0.002
0.108
0.123
0.176
0.003
0.001
0.1
0.009
0.012
0.011
0.042
0.005
0.027
0.146
0.007
0.038
0.001
0.167
0.016
0.026
3.164
0.001
0.111
0.157
0.231
0.001
0.002
0.2
0.009
0.012
0.002
0.033
0.004
0.023
0.142
0.008
0.049
0.001
0.329
0.019
0.091
3.006
0.001
0.159
0.142
0.21
0.004
0.002
0.3
0.009
0.012
0.005
0.046
0.005
0.026
0.137
0.012
0.047
0.001
0.294
0.021
0.024
2.888
0.001
0.067
0.141
0.178
0.006
0.003
0.2
0.009
0.011
0.004
0.042
0.002
0.021
0.107
0.008
0.042
0
0.255
0.022
0.004
2.503
0.001
0.094
0.097
0.206
0.004
0.003
0.103
0.017
0.007
0.002
0.024
0.002
0.017
0.09
0.013
0.016
0
0.213
0.021
0.03
2.091
0
0.063
0.132
0.119
0.006
0.001
0.114
0.007
0.01
0.005
0.025
0.001
0.028
0.08
0.008
0.027
0
0.15
0.013
0.009
3.198
0
0.038
0.083
0.116
0.002
0.001
0.114
0.006
0.006
0.003
0.009
0.002
0.009
0.05
0.018
0.013
0
0.094
0.009
0.024
1.944
0
0.033
0.08
0.122
0.002
0.001
0.147
0.006
0.007
0.013
0.008
0.001
0.009
0.039
0.021
0.012
0
0.078
0.007
0.172
1.348
0
0.031
0.109
0.113
0.012
0.001
0.117
0.01
0.008
0.003
0.027
0.001
0.024
0.071
0.009
0.026
0
0.165
0.008
0.504
2.716
0.001 0.001 0.116 0.156 0.111 0.174 0.226 0.258 0.007 0.005 0.004 0.003 0.091 0.145 0.018 0.011 0.006 0.012 0.003 0.005 0.025 0.065 0.003 0.003 0.015 0.039 0.099 0.157 0.013 0.012 0.022 0.049 0.001 0.001 0.24 0.337 0.025 0.028 0.113 0.007 2.426 3.317 181
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Safety Authority and the author(s), awarded following a tender procedure. The present document is published complying with the transparency principle to which the European Food Safety Authority is subject. It
may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights
of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Supporting publications 2012:EN-232
0.001 0.001 0.001 0.001 0.002 0.002 0.004 0.005 0.005 0.009 0.01 0.012 0.012 0.012 0.014 0.021 0.024 0.026 0.039 0.046 0.046 0.047 0.049 0.131 0.134 0.137 0.142 0.159 0.17 0.21 0.294 III III III III III III III III III III III III III III III III III III III III III III III III III III III III III III III 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 0.09 HQ(IEDI/TTC) TTC (mg/person/day) 0.0042 0.0012 0.006 0.03 0.0024 0.24 0.00012 0.09 0.015 0.054 0.018 0.0144 0.288 0.0072 0.0252 0.1008 0.00144 0.0156 0.234 0.0276 0.0276 0.0282 0.2352 0.07074 0.0804 0.1644 0.0852 0.0954 0.204 0.504 0.1764 Cramer class (ToxTree 2.5) 0.07 0.02 0.1 0.5 0.02 2 0.0005 0.3 0.05 0.1 0.03 0.02 0.4 0.01 0.03 0.08 0.001 0.01 0.1 0.01 0.01 0.01 0.08 0.009 0.01 0.02 0.01 0.01 0.02 0.04 0.01 HQ (IEDI/ADI) exposure (mg/person) Chemical name BENALAXYL MEPTYLDINOCAP METAFLUMIZONE ZOXAMIDE 2,6‐DICHLOROBENZAMIDE CHLORANTRANILIPROLE CADUSAFOS DICAMBA ETOXAZOLE CLOTHIANIDIN HEXYTHIAZOX CYPROCONAZOLE FLUDIOXONIL PROTHIOCONAZOLE FENBUCONAZOLE THIAMETHOXAM TRIEAZOPHOS FENPYROXIMATE METHOXYFENOZIDE DIFENOCONAZOLE SPIRODICLOFEN FLUOPYRAM FLUOPICOLIDE BUPROFEZIN INDOXACARB FLUBENDIAMIDE BIFENTHRIN BIFENAZATE CYPERMETHRIN BOSCALID NOVALURON ADI (JMPR) mg/kg bw Table 23: HI analyses based on ADI or TTC values
0.046667 0.013333 0.066667 0.333333 0.026667 2.666667 0.001333 1 0.166667 0.6 0.2 0.16 3.2 0.08 0.28 1.12 0.016 0.173333 2.6 0.306667 0.306667 0.313333 2.613333 0.786 0.893333 1.826667 0.946667 1.06 2.266667 5.6 1.96 182
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
CHLOPRYRIFOS METHYL CHLOROTHALONIL HALOXYFOP Sum 0.01 0.02 0.0007 Supporting publications 2012:EN-232
0.177 0.36 0.022134 0.295 0.3 0.527 III III III 2.888 0.09 0.09 0.09 1.966667 4 0.245933 37.8426
183
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
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regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
B
IEDI (mg/person)
A
0.6
ADI (mg/kg bw)
Combined actions of chemicals in food through dissimilar modes of action
10
0.5
0.4
0.3
0.2
0.1
0.0
1
0.1
0.01
0.001
0.0001
C
0.6
0.5
HQ
0.4
0.3
0.2
0.1
0.0
1
D
HQ
0.1
0.01
0.001
E
Cumulative HQ
0.0001
3
2
1
ZOXAMIDE
METAFLUMIZONE
BENALAXYL
MEPTYLDINOCAP
CHLORANTRANILIPROLE
CADUSAFOS
26-DICHLOROBENZAMIDE
DICAMBA
ETOXAZOLE
HEXYTHIAZOX
CLOTHIANIDIN
FLUDIOXONIL
PROTHIOCONAZOLE
FENBUCONAZOLE
CYPROCONAZOLE
TRIEAZOPHOS
THIAMETHOXAM
FENPYROXIMATE
SPIRODICLOFEN
METHOXYFENOZIDE
FLUOPYRAM
DIFENOCONAZOLE
BUPROFEZIN
FLUOPICOLIDE
INDOXACARB
BIFENTHRIN
FLUBENDIAMIDE
BIFENAZATE
BOSCALID
CYPERMETHRIN
NOVALURON
CHLOPRYRIFOS METHYL
HALOXYFOP
CHLOROTHALONIL
0
Chemical name
Figure 19: visualisation of HQ values and their contribution to the HI
Supporting publications 2012:EN-232
184
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Each panel in this graph shows a different attribute of the chemicals listed along the bottom x-axis. A:
international estimated daily intakes (IEDI); B: ADI (JMPR), c) HQ (IEDI/ADI), D: HQ shown on a
log y axis; E: cumulative HQ (HI). The solid line in Panel E indicates the HI (at 2.88), the dotted lines
at 1 and 1.88 indicate the critical values of 1 and HI minus 1, respectively. The interpretation of these
values is discussed in the text.
Supporting publications 2012:EN-232
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
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regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
6
5
4
)
C
TT
( 3
Q
H
2
1
0
0
0.1
0.2
0.3
0.4
0.5
0.6
HQ (ADI)
Figure 20: comparison of HQ values based on ADI or TTC values
Supporting publications 2012:EN-232
186
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exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
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regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Chemical name 2,6‐DICHLOROBENZAMIDE exposure (mg/person) ADI (JMPR) mg/kg bw safety factor' (jMPR) NOAEL (mg/kg bw) species effect assay duration (d) HQ(exp/NOAEL) Table 24: PODI (NOAEL) analysis
0.0024 0.02 100 2 rat microscopic changes in the liver 730 0.00002 BENALAXYL 0.0042 0.07 100 6.5 dog atrophy of the seminiferous tubules 365 1.08E‐05 BIFENAZATE 0.0954 0.01 100 1 dog 365 0.00159 BIFENTHRIN 0.0852 0.01 100 1 rats dev tox 0.00142 BOSCALID 0.504 0.04 100 4.4 730 0.001909 BUPROFEZIN 0.07074 0.009 100 0.9 rat 730 0.00131 CADUSAFOS 0.00012 0.0005 100 0.045 rat compensatory haematopoiesis, alteration in urine analysis parameters and liver toxicity increased incidence of tremors in dams during days 10–19 of gestation and increased fetal and litter incidences of hydroureter without hydronephros increased gamma‐glutamyltransferase activity and increased incidences of hepatic eosinophilic foci in male rats increases in the incidence of thyroid F‐cell hypertrophy inhibition of erythrocyte cholinesterase activity CHLOPRYRIFOS METHYL 0.177 0.01 100 1 rat CHLORANTRANILIPROLE 0.24 2 100 158 mice Supporting publications 2012:EN-232
long‐
term inhibition of brain acetylcholinesterase activity 730 and adrenal vacuolation eosinophilic foci accompanied by hepatocellular 548 4.44E‐05 0.00295 2.53E‐05 187
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may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights
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Combined actions of chemicals in food through dissimilar modes of action
CHLOROTHALONIL 0.36 0.02 100 1.8 rat hypertrophy and increased liver weight kidney toxicity CLOTHIANIDIN 0.054 0.1 100 9.7 rat decreased body weight and food consumption CYPERMETHRIN 0.204 0.02 100 2.2 dog severe clinical signs of neurotoxicity CYPROCONAZOLE 0.0144 0.02 100 2.2 rat DICAMBA 0.09 0.3 100 30 rabbit 2‐year study of toxicity and carcinogenicity and the multigeneration reproduction study in rats based on reduced body weight gain and liver toxicity maternal toxicity (behavioural changes) DIFENOCONAZOLE 0.0276 0.01 100 1 ETOXAZOLE 0.015 0.05 100 5.33 dog FENBUCONAZOLE 0.0252 0.03 100 3 rat FENPYROXIMATE 0.0156 0.01 100 1 rat FLUBENDIAMIDE 0.1644 0.02 100 1.7 Supporting publications 2012:EN-232
long‐
term chron
ic 90 730/
multi
gen repro dev tox reduced body‐weight gains, reduced platelet 730 counts and hepatic hypertrophy liver effects (e.g., increases in serum levels of AP 365/9
and triglycerides, absolute and relative liver 0 weights and incidence of centrilobular hepatocyte hypertrophy) NOAEL from 2yr study 730 reductions in body‐weight gain and plasma 730 protein concentration rat/dog effects on the liver (both sexes), kidney, thyroid 730 and hair loss (females) and decreased eosinophil count (males) observed in a 2‐year feeding study in rats, and on the basis of a NOAEL of 100 ppm (equal to 2.2 mg/kg bw per day), based on increased alkaline phosphatase levels, shortened activated prothrombin time and increased liver weights observed in a 1‐year study in dogs 0.003333 9.28E‐05 0.001545 0.000109 0.00005 0.00046 4.69E‐05 0.00014 0.00026 0.001612 188
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may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights
of the authors.
Combined actions of chemicals in food through dissimilar modes of action
FLUDIOXONIL 0.288 0.4 100 37 rat NOAEL from 2yr study FLUOPICOLIDE 0.2352 0.08 100 7.9 mice/ra
ts FLUOPYRAM 0.0282 0.01 100 1.2 rat HALOXYFOP 0.022134 0.0007 100 0.065 rat organ weight increases and gross and 548 microscopic changes in the liver and kidneys in an 18‐month dietary study of toxicity and carcinogenicity in mice, supported by the NOAEL of 8.4 mg/kg bw per day identified on the basis of histopathological changes in the liver and increased kidney weights in a 2‐year dietary study of toxicity and carcinogenicity in rats changes in liver (hepatocellular hypertrophy, 730 eosinophilic foci) low pup body weight multi
gen HEXYTHIAZOX 0.018 0.03 100 3.2 rat INDOXACARB 0.0804 0.01 100 1.1 dog MEPTYLDINOCAP 0.0012 0.02 100 1.6 dog reduced body weight gain in males METAFLUMIZONE 0.006 0.1 100 12 dog clinical signs of poor general state of health, 365 decreased food consumption reduced body‐
weight gain and body‐weight loss, and changes in haematological parameters Supporting publications 2012:EN-232
730 increases in fatty vacuolation of the adrenals in 730 both sexes, the severity of chronic nephritis and the incidence of swollen/withdrawn testes in males erythrocyte damage and the secondary increase 365 in haematopoiesis in the spleen and liver 90 0.00013 0.000496 0.000392 0.005675 9.38E‐05 0.001218 1.25E‐05 8.33E‐06 189
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may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights
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Combined actions of chemicals in food through dissimilar modes of action
METHOXYFENOZIDE 0.234 0.1 100 10 rat/dog effects on erythrocytes plus liver and thyroid hypertrophy in the long‐term study in rats, and 300 ppm, equal to 9.8 mg/kg bw per day, for haematological effects in the 1‐year study in dogs long‐
term(
r)/1yr
(d) 0.00039 NOVALURON 0.1764 0.01 100 1.1 rat 730 0.002673 PROTHIOCONAZOLE 0.0072 0.01 100 5 rat 730 0.000024 SPIRODICLOFEN 0.0276 0.01 100 1.4 dog 365 0.000329 THIAMETHOXAM 0.1008 0.08 100 8.23 dog 90 0.000204 TRIEAZOPHOS 0.00144 0.001 10 0.0125 Human NOEAL from human volunteer study 21 0.00192 ZOXAMIDE 0.03 0.5 100 48 dog reduced body‐weight gain in females 365 1.04E‐05 Sum: 0.030505 Supporting publications 2012:EN-232
erythrocyte damage and secondary splenic and liver changes gross and microscopic changes in the liver and kidneys in a 2‐ year study of toxicity and carcinogenicity adrenal effects in males and females, and increased relative testes weights in males prolonged thromboplastin time 190
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may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights
of the authors.
Combined actions of chemicals in food through dissimilar modes of action
carcinogen mutagen endocrine disrupter reproduction/de
velopment effect acetylcholinester
ase inhibitor neurotoxicant respiratory tract irritant skin irritant eye irritant Table 25: PPDB human health classifications
x nd nd x nd nd nd nd nd x x ? ? ? x x x ? x ? ? ? ? x ? x nd ? ? nd ? ? x x nd x x x ? ? nd nd ? nd nd nd nd nd x nd x nd x nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd x nd yes x x x x x x ? ? nd nd nd nd nd nd yes nd nd nd nd nd nd x nd ? nd nd nd x ? nd ? ? ? x nd x nd ? ? ? ? ? ? ? yes yes ? x nd ? nd x x yes x x ? ? x x x x x x yes yes x x x x x x x x x x x x x nd nd x x x x x x x x x x x yes x x ? ? x x yes x x x x x x x nd x x nd nd x yes x nd x x nd ? x x nd nd nd x x x nd yes x yes yes x x nd ? nd nd x x nd nd yes x yes nd nd ? x nd x x yes x x x x yes x yes x yes x yes yes ? x yes x yes x nd yes yes yes yes nd ? ? x x ? x yes x x x x x ? yes x yes x yes yes ? x yes ? yes x nd yes yes yes nd ? ? x x x Chemical name 2,6‐
DICHLOROBENZAMIDE BENALAXYL BIFENAZATE BIFENTHRIN BOSCALID BUPROFEZIN CADUSAFOS CHLOPRYRIFOS METHYL CHLORANTRANILIPROLE CHLOROTHALONIL CLOTHIANIDIN CYPERMETHRIN CYPROCONAZOLE DICAMBA DIFENOCONAZOLE ETOXAZOLE FENBUCONAZOLE FENPYROXIMATE FLUBENDIAMIDE FLUDIOXONIL FLUOPICOLIDE FLUOPYRAM HALOXYFOP HEXYTHIAZOX INDOXACARB MEPTYLDINOCAP METAFLUMIZONE METHOXYFENOZIDE NOVALURON PROTHIOCONAZOLE SPIRODICLOFEN THIAMETHOXAM Supporting publications 2012:EN-232
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regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TRIEAZOPHOS x nd nd nd yes yes yes yes yes ZOXAMIDE x nd nd x x x nd x yes For each effect, each chemical was classified in the PPDB as ‘yes’: chemical known to cause
a problem (cells filled red); ‘x’: chemical known not to cause a problem (cells filled white);
‘?’: Possibly, status of chemical not identified (cells filled orange); ‘nd’: No data (cells filled
grey). A single value for which a classification as lacking in the PPDB is indicated by a cell
filled black.
Supporting publications 2012:EN-232
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Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 21: HI analysis breakdown by PPDB health issues
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Figure 22: HI analysis, breakdown by PPDB health issues (excluding chemicals negative
for each effect)
Supporting publications 2012:EN-232
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Conclusions and Recommendations
CONCLUSIONS
16.
Conclusions
This project has updated an analysis of the state-of-the-art of mixture toxicology with
emphasis on combinations of chemicals that produce common effects through different modes
of action and mechanisms. The mixture assessment concept of independent action (IA) is
widely accepted as valid for the evaluation of experimentally observed effects of such
combinations.
The overall aim was to propose a cumulative risk assessment (CRA) approach to dealing with
combinations of dissimilarly acting chemicals. A review of current regulatory practice and
guidance revealed that CRA approaches derived from IA are missing. An additional
complicating factor is that application of the IA concept in regulatory practice requires data
that are rarely available. It was therefore of great importance to analyse whether CRA
approaches derived from dose addition can be used as a default also for dealing with
combinations of dissimilarly acting chemicals. This would be viable if it could be
demonstrated that DA is sufficiently conservative also for dissimilarly acting mixtures.
Of central importance was the confirmation from a review of experimental studies that there
is no current example of a situation in which the concept of IA provides an accurate
prediction that is also more conservative than DA.
In the absence of empirical evidence in support of the notion that the use of DA as a default in
CRA is insufficiently conservative, it was necessary to analyse whether IA could theoretically
yield more conservative predictions than DA, and to determine how likely such a situation
would be. By considering the mathematical features of the two prediction concepts, the
quantitative differences between predictions based on DA or IA were analysed in detail.
Prediction differences are driven by the number of chemicals present in the mixture, the slope
of the dose-response relationships of the individual components, the mixture ratio and the
effect level considered during the evaluation. This analysis demonstrated that the differences
that might be expected on the basis of the dose-response characteristics of chemicals
encountered in ecotoxicology are small (less than one order of magnitude) compared with the
uncertainties normally associated with toxicity tests. Although the features of dose-response
relationships with endpoints relevant for mammalian and human toxicology are less well
established, indications are that there are no significant differences.
It is therefore concluded that DA is a sufficiently conservative default concept also for the
evaluation of mixtures of dissimilarly acting chemicals.
This opens up the opportunity to utilize CRA derived from DA also for the assessment of
combinations of dissimilarly acting chemicals in regulatory practice. CRA approaches derived
from DA are well established and have been utilized widely in practice. Their data
requirements can be met in many situations.
Supporting publications 2012:EN-232
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
The overall conclusion of this project is that it is feasible and justified to utilise CRA methods
and tiered framework analyses originally developed for similarly acting mixtures also for
combinations of dissimilarly acting chemicals. There can be one unified approach for dealing
with mixtures in regulatory practice, irrespective of (often presumed) modes of action.
The application of a tiered framework analysis to a selection of case studies has revealed that
the approach is practicable at lower tiers of the analysis, when dealing with comparatively
data-poor situations. It necessitates grouping of chemicals according to common adverse
effects. Viable approaches for grouping have to rely on a sound understanding of the
physiological processes that underlie the induction of adverse effects. It is concluded that the
data necessary to achieve well-founded groupings according to such phenomenological
criteria are rarely available. Groupings for the purpose of creating common assessment groups
therefore have to fall back on more pragmatic criteria, e.g. derived from target organ toxicity.
The application of such approaches to situations that require more refined data sets is
currently complicated by important data gaps. Many of the data gaps can be traced to the
information requirements in relevant pieces of regulation that are not geared towards meeting
the demands of CRA. Lack of such data for the purposes of hazard assessment has a further
impact on the ability of grouping chemicals according to common adverse outcomes.
17.
Identification of data gaps
The review of the literature has identified a data gap of regulatory relevance: Although
reference cases demonstrating the validity of IA for the prediction of experimental mixture
effects have been established with models relevant to ecotoxicology (algal reproduction,
bacterial growth) no such reference case is known for endpoints relevant to mammalian and
human toxicology, neither in vitro nor in vivo. It would be of merit to establish such a
reference case as a proof of principle.
The evaluation of prediction differences between IA and DA could be verified by using a
large database of concentration-response relationships with ecotoxicological endpoints. An
equivalent database with mammalian toxicological endpoints has not yet been assembled, but
would be very valuable to verify the conclusions reached on the basis of ecotoxicological
data.
Another gap that currently hampers the application of CRA approaches in practice is a lack of
empirical data about the chemicals that occur together in environmental samples, food items
and human tissues. Bridging this gap would necessitate analytical efforts that determine
multiple chemicals in one and the same sample.
The information requirements detailed in important pieces of regulation are currently not
geared towards meeting the standards that are necessary for the conduct of higher tier CRA
analyses. This concerns particularly a lack of data for adverse endpoints that are less sensitive
than the endpoints critical for establishing ADIs.
Supporting publications 2012:EN-232
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Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
RECOMMENDATIONS
It is recommended to use a tiered framework analysis with CRA derived from DA also for the
assessment of mixtures of dissimilarly acting chemicals.
At lower tiers of the analysis, all chemicals deemed relevant for the exposure scenario under
investigation should be assessed, irrespective of their presumed modes of action.
At higher tiers, when the risk estimates at lower tiers are deemed unacceptable, chemicals
known not to contribute to a relevant common adverse outcome are excluded from the
analysis. By way of further refining the analysis, criteria for the grouping of chemicals into
common assessment groups should be applied. These criteria should be based on the
chemicals’ capability of affecting a common adverse endpoint.
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
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regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
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Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
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following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
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229
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Appendices
APPENDIX A
Complete list of references returned by search strategy, classified according to relevance to
the project.
APPENDIX B
List of all chemicals tested in all mixture studies.
APPENDIX C
List of all chemicals tested in mixture studies with dual relevance (relevance to both chemical
dissimilarity and low dose issues).
Supporting publications 2012:EN-232
230
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
Glossary / Abbreviations
ADI
Acceptable daily intake
BfR
German Federal Institute for Risk Assessment
BMD
Benchmark dose
BMDL
Lower confidence limit of the BMD
BPAD
Biological pathway altering dose
BPADL
lower confidence limit of the pathway altering dose
BROD
Benzyloxyresorufin O-dealkylase
CA
Concentration Addition, see dose addition
CAG
Cumulative Assessment Group
CES
Critical effect size
CMG
Common Mechanism Group
CRA
Cumulative risk assessment
DA
Dose Addition, see concentration addition
DA
Dose additivity
DON
Deoxynivalenol
EC
European Commission
EFSA
European Food Safety Authority
EPA
Environmental Protection Agency, United States
EROD
Ethoxyresorufin O-dealkylase
EU
European Union
GEMS/Food Global Environment Monitoring System – Food Contamination
Monitoring and Assessmemt Programme
Hg
Mercuric
HI
Hazard index
HQ
Hazard quotient
IA
Independent action
IPR
Isoproterenol chloride
JECFA
Joint FAO/WHO Expert Committee on Food Additives
JMPR
Joint FAO/WHO Meeting on Pesticide Residues
Supporting publications 2012:EN-232
231
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
LEL
Lowest effect level
LOAEL
Lowest observed adverse effect level
LOD
Limit of detection
LOQ
Limit of quantification
MRL
Maximum residue level
mRNA
Messenger Ribonucleic acid
NAS
National Academy of Science
NEL
No effect level
NIV
Nivalenol
NOAEL
No observed adverse effect level
NOEL
No observed effect level
NOTEL
No observed transcription effect level
OECD
Organisation for Economic Co-operation and Development
PB
Phenobarbital
PbAc
Lead acetate
PCB 126
3,3’,4,4’,5-Pentachlorobiphenyle
POD
Point of departure
PPP
Plant protection product
PRIMo
Pesticide Residue Intake Model
PROD
Pentoxyresorufin O-dealkylase
RAC
Raw Agricultural Commodities
RfD
Reference dose
SCF
Scientific Committee on Food
SCOOP
Scientific cooperation tasks
T3
Triiodothyronine
T4
Tetraiodothyronine
TCDD
Tetrachlorodibenzodioxin
TDI
Tolerable daily intake
TEF
Toxic Equivalency Factor
TEQ
Toxic Equivalency Quotient
Supporting publications 2012:EN-232
232
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Combined actions of chemicals in food through dissimilar modes of action
TSH
Thyroid stimulating hormone
tTDI
Temporary tolerable daily intake
UK
United Kingdom
WBC
White blood cell
WHO
World Health Organization
Supporting publications 2012:EN-232
233
The present document has been produced and adopted by the bodies identified above as author(s). This task has been carried out
exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the author(s), awarded
following a tender procedure. The present document is published complying with the transparency principle to which the European Food
Safety Authority is subject. It may not be considered as an output adopted by EFSA. EFSA reserves its rights, view and position as
regards the issues addressed and the conclusions reached in the present document, without prejudice to the rights of the authors.
Appendices
Investigation of the state of the science on combined actions of chemicals in
food through dissimilar modes of action and proposal for science-based
approach for performing related cumulative risk assessment
CFT/EFSA/PPR/2010/04
APPENDIX A
Complete list of references returned by search strategy, classified according to relevance to
the project.
APPENDIX B
List of all chemicals tested in all mixture studies.
APPENDIX C
List of all chemicals tested in mixture studies with dual relevance (relevance to both chemical
dissimilarity and low dose issues).
APPENDIX A
Page 1 of 66
CFT/EFSA/PPR/2010/02
Classification of IA search results.accdb
Results sorted by Authors field
Authors
PubYear
ID
Title
Relevance
Abdelkhalek MB;Breton MF;Feliers D;Haye
B;Pavlovic-Hournac M;
1994
1054
TSH action on cAMP binding to the regulatory subunits of
ON trivial
cAMP-dependent protein kinases in pig thyroid cell cultures
Abend WK;
1978
62
RESPONSE TO CONSTANT ANGULAR ACCELERATIONS OF
NEURONS IN MONKEY SUPERIOR VESTIBULAR NUCLEUS
OFF domain
Aboudy Y;Mendelson E;Shalit I;Bessalle
R;Fridkin M;
1994
941
ACTIVITY OF 2 SYNTHETIC AMPHIPHILIC PEPTIDES AND
MAGAININ-2 AGAINST HERPES-SIMPLEX VIRUS TYPE-1
AND TYPE-2
OFF domain
Acs Z;Lonart G;Makara GB;
1990
294
ROLE OF HYPOTHALAMIC FACTORS (GROWTHHORMONE-RELEASING HORMONE AND GAMMAAMINOBUTYRIC-ACID) IN THE REGULATION OF
GROWTH-HORMONE SECRETION IN THE NEONATAL AND
ADULT-RAT
OFF domain
Adachi K;Yoshikawa K;Halprin KM;Levine V;
1975
34
PROSTAGLANDINS AND CYCLIC-AMP IN EPIDERMIS EVIDENCE FOR INDEPENDENT ACTION OF
PROSTAGLANDINS AND ADRENALIN ON ADENYLCYCLASE SYSTEM OF PIG AND HUMAN EPIDERMIS,
NORMAL AND PSORIATIC
OFF domain
Afonso SG;de Salamanca RE;Batlle A;
2001
464
Photodynamic and light independent action of 8 to 2
carboxylic free porphyrins on some haem-enzymes
OFF domain
Agullo L;Garcia-Dorado D;Inserte J;Paniagua
A;Pyrhonen P;Llevadot J;Soler-Soler J;
1999
984
L-Arginine limits myocardial cell death secondary to
hypoxia-reoxygenation by a cGMP-dependent mechanism
OFF domain
Ahmed MI;Gaddam KK;Gladden JD;Desai
RV;Lloyd SG;Gupta H;Denney TS;Oparil
S;Dell'Italia LJ;Calhoun DA;
2009
344
Spironolactone Therapy is Associated With Rapid
Improvements in Blood Pressure Control and Cardiac
Remodeling in Patients With Resistant Hypertension
OFF domain
Ahren B;Lindskog S;Vandijk G;Scheurink
AJW;Steffens AB;
1995
1028
EFFECTS OF GLP-1 AND 2,5-ANHYDRO-D-MANNITOL ON
INSULIN-SECRETION AND PLASMA-GLUCOSE IN MICE
OFF domain
Ali S;Becker MW;Davis MG;Dorn GW;
1994
1060
DISSOCIATION OF VASOCONSTRICTOR-STIMULATED
BASIC FIBROBLAST GROWTH-FACTOR EXPRESSION
FROM HYPERTROPHIC GROWTH IN CULTURED
VASCULAR SMOOTH-MUSCLE CELLS - RELEVANT ROLES
OF PROTEIN-KINASE-C
OFF domain
Aliapoulios MA;Hattner RS;Bernstein
MD;George B;Rose EH;
1969
10
MODE OF INDEPENDENT ACTION OF GLUCAGON
HORMONE METAB AND CALCITONIN HORMONE METAB
OFF domain
Alila HW;Corradino RA;Hansel W;
1990
295
ARACHIDONIC-ACID AND ITS METABOLITES INCREASE
CYTOSOLIC FREE CALCIUM IN BOVINE LUTEAL CELLS
OFF domain
Allen JM;Bircham PMM;Edwards AV;Tatemoto
K;Bloom SR;
1983
149
NEUROPEPTIDE Y (NPY) REDUCES MYOCARDIAL
PERFUSION AND INHIBITS THE FORCE OF
CONTRACTION OF THE ISOLATED PERFUSED RABBIT
HEART
OFF domain
Allen R;
2007
880
Criminal justice and women, 1792-1811 (French
Revolution)
OFF domain
Altenburger R;Backhaus T;Boedeker W;Faust
M;Scholze M;Grimme LH;
2000
992
Predictability of the toxicity of multiple chemical mixtures
to Vibrio fischeri: Mixtures composed of similarly acting
chemicals
ON experimental
study
Altenburger R;Boedeker W;Faust M;Grimme
LH;
1996
605
Regulations for combined effects of pollutants:
Consequences from risk assessment in aquatic toxicology
ON review
Altenburger R;Schmitt H;Schuurmann G;
2005
416
Algal toxicity of nitrobenzenes: Combined effect analysis
ON experimental
as a pharmacological probe for similar modes of interaction study
Altenburger R;Walter H;Grote M;
2004
894
What contributes to the combined effect of a complex
mixture?
ON review
Alto LE;Dhalla NS;
1979
74
MYOCARDIAL CATION CONTENTS DURING INDUCTION
OF CALCIUM PARADOX
OFF domain
Alvarez G;
1982
129
THE NEUROLOGY OF POVERTY
OFF domain
APPENDIX A
Authors
Page 2 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Amano Y;
1982
930
WINTER WHEAT BREEDING FOR RESISTANCE TO SNOW
MOLD AND COLD HARDINESS 2. ESTIMATION OF
GENETIC EFFECTS FOR RESISTANCE TO SNOW MOLD
AND FREEZING BY THE USE OF HALF DIALLEL CROSS
ANALYSIS
OFF domain
Andersen HU;Jorgensen KH;Egeberg
J;Mandruppoulsen T;Nerup J;
1994
705
NICOTINAMIDE PREVENTS INTERLEUKIN-1 EFFECTS ON
ACCUMULATED INSULIN RELEASE AND NITRIC-OXIDE
PRODUCTION IN RAT ISLETS OF LANGERHANS
OFF domain
Andersen IS;Voie OA;Fonnum F;Mariussen E;
2009
798
Effects of Methyl Mercury in Combination with
Polychlorinated Biphenyls and Brominated Flame
Retardants on the Uptake of Glutamate in Rat Brain
Synaptosomes: A Mathematical Approach for the Study of
Mixtures
ON experimental
study
Anderson KM;Wygodny JB;Ondrey F;Harris J;
1988
248
HUMAN PC-3 PROSTATE CELL-LINE DNA-SYNTHESIS IS
SUPPRESSED BY EICOSATETRAYNOIC ACID, AN INVITRO
INHIBITOR OF ARACHIDONIC-ACID METABOLISM
OFF domain
Andersson M;Low P;Bakhiet M;
1996
639
Lovastatin inhibits interferon-gamma-induced
Trypanosoma brucei brucei proliferation: Evidence for
mevalonate pathway involvement
OFF domain
Ando B;Wiedmer T;Sims PJ;
1989
280
THE SECRETORY RELEASE REACTION INITIATED BY
OFF domain
COMPLEMENT PROTEINS-C5B-9 OCCURS WITHOUT
PLATELET-AGGREGATION THROUGH GLYCOPROTEIN-IIBIIIA
Aniksztejn L;Catarsi S;Drapeau P;
1997
365
Channel modulation by tyrosine phosphorylation in an
identified leech neuron
OFF domain
Antognini JF;Raines DE;Solt K;Barter
LS;Atherley RJ;Bravo E;Laster MJ;Jankowska
K;Eger EI;
2007
462
Hexafluorobenzene acts in the spinal cord, whereas odifluorobenzene acts in both brain and spinal cord, to
produce immobility
OFF domain
Antonello JM;Raghavarao D;
2000
638
Optimal designs for the individual and joint exposure
general logistic regression models
ON method dev.
Aoyagi K;Ohhara N;Okamura S;Otsuka
T;Shibuya T;Yamano Y;Tsuda Y;Niho Y;
1987
227
APLASTIC-ANEMIA ASSOCIATED WITH TYPE-A VIRALHEPATITIS - POSSIBLE ROLE OF LYMPHOCYTES-T
OFF domain
Apa R;Lanzone A;Miceli F;Caruso A;Mancuso
S;Canipari R;
1994
990
GROWTH-HORMONE INDUCTION OF RAT GRANULOSACELL TISSUE-PLASMINOGEN ACTIVATOR EXPRESSION
AND PROGESTERONE SYNTHESIS
OFF domain
Appenroth KJ;Augsten H;Mohr H;
1992
659
PHOTOPHYSIOLOGY OF TURION GERMINATION IN
SPIRODELA-POLYRHIZA (L) SCHLEIDEN .10. ROLE OF
NITRATE IN THE PHYTOCHROME-MEDIATED RESPONSE
OFF domain
Araki S;
2009
962
Independent action of the Japanese Society of Neurology
is a driving force behind the development of Japanese
neurology
OFF domain
Arimilli S;Johnson JB;exander-Miller MA;Parks
GD;
2007
626
TLR-4 and -6 agonists reverse apoptosis and promote
maturation of simian virus 5-infected human dendritic cells
through NF kappa B-dependent
OFF domain
Armitage JP;Gallagher A;Johnston AWB;
1988
249
COMPARISON OF THE CHEMOTACTIC BEHAVIOR OF
RHIZOBIUM LEGUMINOSARUM WITH AND WITHOUT THE
NODULATION PLASMID
OFF domain
Arrhenius A;Backhaus T;Gronvall F;Junghans
M;Scholze M;Blanck H;
2006
1026
Effects of three antifouling agents on algal communities
and algal reproduction: Mixture toxicity studies with TBT,
Irgarol, and Sea-Nine
ON experimental
study
Arrhenius A;Gronvall F;Scholze M;Backhaus
T;Blanck H;
2004
835
Predictability of the mixture toxicity of 12 similarly acting
congeneric inhibitors of photosystem II in marine
periphyton and epipsammon communities
ON experimental
study
Arutyunyan RS;Lenkov DN;
1983
666
MODULATION OF MUSCLE AFFERENTS BY MOTOR AND
SUPPLEMENTARY MOTOR CORTEX STIMULATION IN THE
CAT
OFF domain
Asa SL;Gerrie BM;Kovacs K;Horvath E;Singer
W;Killinger DW;Smyth HS;
1988
250
STRUCTURE-FUNCTION CORRELATIONS OF HUMAN
PITUITARY GONADOTROPH ADENOMAS INVITRO
OFF domain
APPENDIX A
Authors
Page 3 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Asai T;Shuba LM;Pelzer DJ;McDonald TF;
1996
395
PKC-independent inhibition of cardiac L-type Ca-2+
channel current by phorbol esters
OFF domain
Asem EK;Tsang BK;
1988
251
THE EFFECT OF KAURENOL ON STEROIDOGENESIS AND
CYCLIC ADENOSINE-MONOPHOSPHATE PRODUCTION IN
RAT GRANULOSA-CELLS
OFF domain
Asp NG;Agardh CD;Ahren B;Dencker
I;Johansson CG;Lundquist I;Nyman M;Sartor
G;Schersten B;
1981
791
Dietary fibre in type II diabetes
OFF domain
Aydin A;Pearce JM;
1997
802
Some determinants of response summation
OFF domain
B. E. Korba;P. Cote;W. Hornbuckle;R.
Schinazi;J. D. Gangemi;B. C. Tennant;J. L.
Gerin
2000
1142
Enhanced antiviral benefit of combination therapy with
lamivudine and alpha interferon against WHV replication in
chronic carrier woodchucks
ON trivial
B. E. Korba;P. Cote;W. Hornbuckle;R.
Schinazi;J. L. Gerin;B. C. Tennant
2000
1143
Enhanced antiviral benefit of combination therapy with
lamivudine and famciclovir against WHV replication in
chronic WHV carrier woodchucks
ON trivial
B. Seigneres;P. Martin;B. Werle;O. Schorr;C.
Jamard;L. Rimsky;C. Trepo;F. Zoulim
2003
1152
Effects of pyrimidine and purine analog combinations in
the duck hepatitis B virus infection model
ON experimental
study
Babu RV;Anantharaman B;Ramakrishnan
KR;Srinivasan SH;
2002
1048
Compressed domain action classification using HMM
OFF domain
Backhaus T;Altenburger R;Arrhenius A;Blanck
H;Faust M;Finizio A;Gramatica P;Grote
M;Junghans M;Meyer W;Pavan M;Porsbring
T;Scholze M;Todeschini R;Vighi M;Walter
H;Grimme LH;
2003
552
The BEAM-project: prediction and assessment of mixture
toxicities in the aquatic environment
ON experimental
study
Backhaus T;Altenburger R;Boedeker W;Faust
M;Scholze M;Grimme LH;
2000
389
Predictability of the toxicity of a multiple mixture of
dissimilarly acting chemicals to Vibrio fischeri
ON experimental
study
Backhaus T;Arrhenius A;Blanck H;
2004
761
Toxicity of a mixture of dissimilarly acting substances to
natural algal communities: Predictive power and
limitations of independent action and concentration
addition
ON experimental
study
Backhaus T;Faust M;Scholze M;Gramatica
P;Vighi M;Grimme LH;
2004
750
Joint algal toxicity of phenylurea herbicides is equally
predictable by concentration addition and independent
action
ON experimental
study
Backhaus T;Scholze M;Grimme LH;
2000
784
The single substance and mixture toxicity of quinolones to
the bioluminescent bacterium Vibrio fischeri
ON experimental
study
Badr FM;El-Habit OHM;Hamdy M;Hassan GAR;
1998
995
The mutagenic versus protective role of vitamin A on the
OFF domain
induction of chromosomal aberration in human lymphocyte
cultures
Balakrishnan S;Pandhi P;Bhargava VK;
1998
524
Effect of nimodipine on the efficacy of commonly used
antiepileptic drugs in rats
ON experimental
study
Banaiee N;Jacobs WR;Ernst JD;
2007
733
LspA-independent action of globomycin on Mycobacterium
tuberculosis
OFF domain
Bankar GR;Nampurath GK;Nayak
PG;Bhattacharya S;
2010
1107
A possible correlation between the correction of
endothelial dysfunction and normalization of high blood
pressure levels by 1,3,4-oxadiazole derivative, an L-type
Ca2+ channel blocker in deoxycorticosterone acetate and
N-G-nitro-L-arginine hypertensive ra
OFF domain
Barata C;Baird DJ;Nogueira AJA;Agra
AR;Soares A;
2007
1079
Life-history responses of Daphnia magna Straus to binary
mixtures of toxic substances: Pharmacological versus
ecotoxicological modes of action
ON experimental
study
Barata C;Baird DJ;Nogueira AJA;Soares
A;Riva MC;
2006
1077
Toxicity of binary mixtures of metals and pyrethroid
insecticides to Daphnia magna Straus. Implications for
multi-substance risks assessment
ON experimental
study
Barmaz S;Potts SG;Vighi M;
2010
756
A novel method for assessing risks to pollinators from
plant protection products using honeybees as a model
species
ON experimental
study
Barnett P;Kruitbosch DL;Hemrika W;Dekker
HL;Wever R;
1997
906
The regulation of the vanadium chloroperoxidase from
Curvularia inaequalis
OFF domain
APPENDIX A
Authors
Page 4 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Baronas-Lowell D;Lauer-Fields JL;Fields GB;
2004
531
Induction of endothelial cell activation by a triple helical
alpha(2)beta(1) integrin ligand, derived from type I
collagen alpha 1(I)496-507
OFF domain
Barteneva OI;Bartenev VV;
2004
771
Inhibitive effect of metal salts on aluminum corrosion in
hydrochloride electrolytes
OFF domain
Barton CN;Braunberg RC;Friedman L;
1993
378
NONLINEAR STATISTICAL-MODELS FOR THE JOINT
ACTION OF TOXINS
ON method dev.
Basham C;Mills J;Douple EB;Roberts JJ;
1988
252
INDEPENDENT ACTION OF RADIATION AND PLATINUM
COMPOUNDS ON HUMAN-FETAL LUNG-CELLS INVITRO
AND HUMAN-MELANOMA CELLS INVIVO
ON trivial
Basham C;Mills J;Douple EB;Roberts JJ;
1989
281
THE INDEPENDENT ACTION OF RADIATION AND
CISPLATIN ON THE SURVIVAL OR RECOVERY OF HUMAN
NORMAL OR TUMOR-CELLS INVITRO OR INVIVO
OFF domain
Beaumont K;
1987
228
ADRENOCORTICOID RECEPTORS IN C6 GLIOMA-CELLS EFFECTS ON CELL-GROWTH
OFF domain
Beck B;
1986
823
DEGREE OF REALITY OF SELF-ASSESSMENTS OF HARD
OF HEARING AND DEAF SCHOOL AGE CHILDREN
OFF domain
Beeckmans S;Kanarek L;
1983
150
THE MODIFICATION WITH TETRANITROMETHANE OF AN
ESSENTIAL TYROSINE IN THE ACTIVE-SITE OF PIG
FUMARASE
OFF domain
Beetham KL;Busse PM;Tolmach LJ;
1983
151
SYNERGISTIC KILLING OF HELA-CELLS BY
HYDROXYUREA AND CAFFEINE
OFF domain
Behan WMH;McDonald M;Darlington
LG;Stone TW;
1999
355
Oxidative stress as a mechanism for quinolinic acidinduced hippocampal damage: protection by melatonin
and deprenyl
OFF domain
Belden JB;Gilliom RJ;Lydy MJ;
2007
804
How well can we predict the toxicity of pesticide mixtures
to aquatic life?
ON review
Belden JB;Lydy MJ;
2006
968
Joint toxicity of chlorpyrifos and esfenvalerate to fathead
minnows and midge larvae
ON experimental
study
Bell DA;Peeters GA;Davis WL;Kohmoto
O;Nelson JA;Barry WH;
1988
253
EFFECTS OF CONTRAST-MEDIA ON CALCIUM
TRANSIENTS AND MOTION IN CULTURED VENTRICULAR
CELLS
OFF domain
Bellas J;
2008
892
Prediction and assessment of mixture toxicity of
compounds in antifouling paints using the sea-urchin
embryo-larval bioassay
ON experimental
study
Benabdelkhalek M;Breton MF;Feliers D;Haye
B;Pavlovichournac M;
1994
1055
TSH ACTION ON CAMP BINDING TO THE REGULATORY
SUBUNITS OF CAMP-DEPENDENT PROTEIN-KINASES IN
PIG THYROID-CELL CULTURES
OFF domain
Benigni R;Richard AM;
1996
1036
QSARS of mutagens and carcinogens: Two case studies
illustrating problems in the construction of models for
noncongeneric chemicals
ON review
Benmenahem D;Shragalevine Z;Limor R;Naor
Z;
1994
1008
ARACHIDONIC-ACID AND LIPOXYGENASE PRODUCTS
STIMULATE GONADOTROPIN ALPHA-SUBUNIT
MESSENGER-RNA LEVELS IN PITUITARY ALPHA-T3-1
CELL-LINE - ROLE IN GONADOTROPIN-RELEASINGHORMONE ACTION
OFF domain
Benning CM;Kyprianou N;
2002
789
Quinazoline-derived alpha 1-adrenoceptor antagonists
induce prostate cancer cell apoptosis via an alpha 1adrenoceptor-independent action
OFF domain
Bensch S;Grahn M;Muller N;Gay L;Akesson S;
2009
873
Genetic, morphological, and feather isotope variation of
migratory willow warblers show gradual divergence in a
ring
OFF domain
Ben-Yosef G;Ben-Shahar O;
2008
793
Curvature-based perceptual singularities and texture
saliency with early vision mechanisms
OFF domain
Benz G;
1975
35
ACTION OF BACILLUS-THURINGIENSIS PREPARATION
AGAINST LARCH BUD MOTH, ZEIRAPHERA-DINIANA
(GN), ENHANCED BY BETA-EXOTOXIN AND DDT
OFF domain
APPENDIX A
Authors
Page 5 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Berdicev.I;Grossowi.N;
1972
21
REVERSAL BY CALCIUM-IONS OF GROWTH INHIBITION
OF DEBARYOMYCES-NICOTIANAE CAUSED BY
ANTIFUNGAL POLYENE ANTIBIOTICS
OFF domain
Berschneider HM;Blikslager AT;Roberts MC;
1999
561
Role of duodenal reflux in nonglandular gastric ulcer
disease of the mature horse
OFF domain
Bertrand I;Gayethallion T;Quivy D;
1950
1
ETUDE PHARMACODYNAMIQUE DE LACTION CONJOINTE
INDEPENDANTE DE 2 SPASMOLYTIQUES - PYRAMIDON
ET DI-IODO SALIGENINE
OFF domain
Bhattacharya S;Banerjee A;Bandyopadhyay S;
2005
415
CORBA-based analysis of multi agent behavior
OFF domain
Bhattacharya SK;Tripathi SR;Pradhan
CK;Kashyap SK;
1990
296
ACUTE EFFECTS OF HEAT ON NEUROPSYCHOLOGICAL
CHANGES AND PHYSIOLOGICAL-RESPONSES UNDER
NOISE CONDITION
OFF domain
Bielefeld DR;Pace CS;Boshell BR;
1983
152
ALTERED SENSITIVITY OF CHRONIC DIABETIC RATHEART TO CALCIUM
OFF domain
Binder N;Landon EJ;Wecker L;Dettbarn WD;
1976
44
EFFECT OF PARATHION AND ITS METABOLITES ON
CALCIUM-UPTAKE ACTIVITY OF RAT SKELETAL-MUSCLE
SARCOPLASMIC-RETICULUM INVITRO
OFF domain
Binoux M;Hossenlopp P;Gourmelen M;Girard
F;
1980
85
BIOSYNTHESIS AND HORMONAL-REGULATION OF IGF
(INSULIN-LIKE GROWTH-FACTORS, OR
SOMATOMEDINS) - EXPERIMENTAL AND CLINICALSTUDIES
OFF domain
Bird SP;Tarpenning KM;Marino FE;
2006
387
Liquid carbohydrate/essential amino acid ingestion during
a short-term bout of resistance exercise suppresses
myofibrillar protein degradation
OFF domain
Black VJ;Ormrod DP;Unsworth MH;
1982
130
EFFECTS OF LOW CONCENTRATION OF OZONE, SINGLY,
AND IN COMBINATION WITH SULFUR-DIOXIDE ON NET
PHOTOSYNTHESIS RATES OF VICIA-FABA L
ON trivial
Blau M;Thompson G;
1991
1110
DO METRIC INDEPENDENT CLASSICAL ACTIONS LEAD
TO TOPOLOGICAL FIELD-THEORIES
OFF domain
Blum JJ;
1992
663
ARGININE CATABOLISM BY LEISHMANIA-DONOVANI
PROMASTIGOTES
OFF domain
Bode HP;Moormann B;Dabew R;Goke B;
1999
566
Glucagon-like peptide 1 elevates cytosolic calcium in
pancreatic beta-cells independently of protein kinase A
OFF domain
Boedeker W;Backhaus T;
2010
859
The scientific assessment of combined effects of risk
factors: different approaches in experimental biosciences
and epidemiology
ON review
Boehmer S;Carapito C;Wilzewski B;Leize
E;Van Dorsselaer A;Bernhardt R;
2006
608
Analysis of aldosterone-induced differential receptorOFF domain
independent protein patterns using 2D-electrophoresis and
mass spectrometry
Boelema BH;
1973
25
INFECTIVITY TITRATIONS WITH SOME PLANT
PATHOGENIC BACTERIA
OFF domain
Boelema BH;
1985
182
DOSE-RESPONSE RELATIONSHIPS BETWEEN
PHYTOPATHOGENIC BACTERIA AND THEIR HOSTS
OFF domain
Bollema BH;
1984
889
INFECTIVITY TITRATIONS WITH RACE 2 OF
PSEUDOMONAS-SYRINGAE PATHOVAR PHASEOLICOLA IN
GREEN BEANS PHASEOLUS-VULGARIS
OFF domain
Bolt HM;Mumtaz MM;
1996
465
Risk assessment of mixtures and standard setting:
Working towards practical compromises
ON review
Borgert CJ;Quill TF;McCarty LS;Mason AM;
2004
790
Can mode of action predict mixture toxicity for risk
assessment?
ON review
Borgmann U;Norwood WP;Dixon DG;
2008
471
Modelling bioaccumulation and toxicity of metal mixtures
ON experimental
study
Boroojerdi B;Ferbert A;Foltys H;Kosinski
CM;Noth J;Schwarz M;
1999
728
Evidence for a non-orthostatic origin of orthostatic tremor
OFF domain
Boucher AN;Tam VH;
2006
841
Mathematical formulation of additivity for antimicrobial
agents
ON experimental
study
APPENDIX A
Authors
Page 6 of 66
PubYear
ID
CFT/EFSA/PPR/2010/02
Title
Relevance
Bourguet J;Morel F;
1967
8
INDEPENDANCE DES VARIATIONS DE PERMEABILITE A
LEAU ET AU SODIUM PRODUITES PAR LES HORMONES
NEUROHYPOPHYSAIRES SUR LA VESSIE DE GRENOUILLE
OFF domain
Bowen WH;Pearson SK;Falany JL;
1990
297
INFLUENCE OF SWEETENING AGENTS IN SOLUTION ON
DENTAL-CARIES IN DESALIVATED RATS
OFF domain
Bowers CY;Sartor AO;Reynolds GA;Badger
TM;
1991
562
ON THE ACTIONS OF THE GROWTH HORMONERELEASING HEXAPEPTIDE, GHRP
OFF domain
Bowman AM;Nesin OM;Pakhomova
ON;Pakhomov AG;
2010
922
Analysis of Plasma Membrane Integrity by Fluorescent
Detection of Tl+ Uptake
OFF domain
Braun M;Wendt A;Buschard K;Salehi
A;Sewing S;Gromada J;Rorsman P;
2004
1105
GABA(B) receptor activation inhibits exocytosis in rat
pancreatic beta-cells by G-protein-dependent activation of
calcineurin
OFF domain
Brigotti M;Petronini PG;Carnicelli D;Alfieri
RR;Bonelli MA;Borghetti AF;Wheeler KP;
2003
1068
Effects of osmolarity, ions and compatible osmolytes on
cell-free protein synthesis
OFF domain
Brill A;Elinav H;Varon D;
2004
732
Differential role of platelet granular mediators in
angiogenesis
OFF domain
Brinkmann V;Kristofic C;
1995
654
TCR-STIMULATED NAIVE HUMAN CD4(+)45R0(-) T-CELLS OFF domain
DEVELOP INTO EFFECTOR-CELLS THAT SECRETE IL-13,
IL-5, AND IFN-GAMMA, BUT NO IL-4, AND HELP
EFFICIENT IGE PRODUCTION BY B-CELLS
Brock B;Mogensen JH;Gregersen
S;Hermansen K;
2002
1082
Glucose desensitization in INS-1 cells: Evidence of
impaired function caused by glucose metabolite(s) rather
than by the glucose molecule per se
OFF domain
Broderius SJ;Kahl MD;Elonen
GE;Hammermeister DE;Hoglund MD;
2005
966
A comparison of the lethal and sublethal toxicity of organic
chemical mixtures to the fathead minnow (Pimephales
promelas)
ON experimental
study
Broderius SJ;Kahl MD;Hoglund MD;
1995
418
USE OF JOINT TOXIC RESPONSE TO DEFINE THE
PRIMARY-MODE OF TOXIC ACTION FOR DIVERSE
INDUSTRIAL ORGANIC-CHEMICALS
ON experimental
study
Broderius SJ;Smith LL;
1979
75
LETHAL AND SUBLETHAL EFFECTS OF BINARY-MIXTURES
OF CYANIDE AND HEXAVALENT CHROMIUM, ZINC, OR
AMMONIA TO THE FATHEAD MINNOW (PIMEPHALESPROMELAS) AND RAINBOW-TROUT (SALMO-GAIRDNERI)
OFF domain
Brody S;Yanofsky C;
1964
6
INDEPENDENT ACTION OF ALLELE-SPECIFIC
SUPPRESSOR MUTATIONS
OFF domain
Broeders MAW;Tangelder GJ;Slaaf
DW;Reneman RS;Egbrink M;
1998
466
Endogenous nitric oxide protects against
thromboembolism in venules but not in arterioles
OFF domain
Broerse M;van Gestel CAM;
2010
1080
Mixture effects of nickel and chlorpyrifos on Folsomia
candida (Collembola) explained from development of
toxicity in time
ON experimental
study
Brooks J;
1997
886
Congressional voting on farm payment limitations: Political
pressure or ideological conviction?
OFF domain
Brosche S;Backhaus T;
2010
430
Toxicity of five protein synthesis inhibiting antibiotics and
their mixture to limnic bacterial communities
ON experimental
study
Brotherton AFA;Macfarlane DE;Hoak JC;
1982
131
PROSTACYCLIN BIOSYNTHESIS IN VASCULAR
ENDOTHELIUM IS NOT INHIBITED BY CYCLIC-AMP STUDIES WITH 3-ISOBUTYL-1-METHYLXANTHINE AND
FORSKOLIN
OFF domain
Bruch P;Schnackerz KD;Gracy RW;
1976
45
MATRIX-BOUND PHOSPHOGLUCOSE ISOMERASE FORMATION AND PROPERTIES OF PROPERTIES OF
MONOMERS AND HYBRIDS
OFF domain
Buccafusco JJ;Heithold DL;Chon SH;
1990
298
LONG-TERM BEHAVIORAL AND LEARNING
ABNORMALITIES PRODUCED BY THE IRREVERSIBLE
CHOLINESTERASE INHIBITOR SOMAN - EFFECT OF A
STANDARD PRETREATMENT REGIMEN AND CLONIDINE
OFF domain
APPENDIX A
Authors
Page 7 of 66
PubYear
ID
CFT/EFSA/PPR/2010/02
Title
Relevance
Bunda S;Wang YT;Mitts TF;Liu P;Arab
S;Arabkhari M;Hinek A;
2009
1114
Aldosterone Stimulates Elastogenesis in Cardiac
OFF domain
Fibroblasts via Mineralocorticoid Receptor-independent
Action Involving the Consecutive Activation of G alpha(13),
c-Src, the Insulin-like Growth Factor-I Receptor, and
Phosphatidylinositol 3-Kinase/Akt
Buntzen S;Nordgren S;Delbro D;Hulten L;
1995
959
REFLEX INTERACTION FROM THE URINARY-BLADDER
AND THE RECTUM ON ANAL MOTILITY IN THE CAT
OFF domain
Burstein SA;Adamson JW;Harker LA;
1980
86
MEGAKARYOCYTOPOIESIS IN CULTURE - MODULATION
BY CHOLINERGIC MECHANISMS
OFF domain
Buxton ILO;Brunton LL;
1985
183
ACTION OF THE CARDIAC ALPHA-1-ADRENERGIC
OFF domain
RECEPTOR - ACTIVATION OF CYCLIC-AMP DEGRADATION
Bychkov SM;Kuz'Mina SA;
1977
1001
SIMULTANEOUS ACTION OF PROTEIN CHONDROITIN 4
KERATO SULFATE AND HYALURONIC-ACID ON THE
AGGREGATION AND ADHESION OF ERYTHROCYTES
OFF domain
Cabrefiga J;Montesinos E;
2005
848
Analysis of aggressiveness of Erwinia amylovora using
disease-dose and time relationships
OFF domain
Cabrerajuarez E;Espinosalara M;
1990
299
MUTAGENIC AND LETHAL ACTION OF POLYCHROMATIC
NEAR-ULTRAVIOLET (325-400 NM) ON HAEMOPHILUSINFLUENZAE IN THE PRESENCE OF NITROGEN
OFF domain
Candipan RC;Sjostrand FS;
1984
162
WATER-MOVEMENT FROM INTRACRYSTAL SPACES IN
ISOLATED LIVER-MITOCHONDRIA
OFF domain
Caratsch CG;Maranda B;Miledi R;Strong PN;
1981
106
A FURTHER STUDY OF THE PHOSPHOLIPASEINDEPENDENT ACTION OF BETA-BUNGAROTOXIN AT
FROG ENDPLATES
OFF domain
Caratsch CG;Miledi R;Strong PN;
1985
184
INFLUENCE OF DIVALENT-CATIONS ON THE
OFF domain
PHOSPHOLIPASE-INDEPENDENT ACTION OF BETABUNGAROTOXIN AT FROG NEUROMUSCULAR-JUNCTIONS
Cardenas M;Schillen K;Pebalk D;Nylander
T;Lindman B;
2005
955
Interaction between DNA and charged colloids could be
hydrophobically driven
OFF domain
Cardoso CMP;Custodio JBA;Almeida
LM;Moreno AJM;
2001
833
Mechanisms of the deleterious effects of tamoxifen on
mitochondrial respiration rate and phosphorylation
efficiency
OFF domain
Carlberg C;
1993
324
RXR-INDEPENDENT ACTION OF THE RECEPTORS FOR
THYROID-HORMONE, RETINOID ACID AND VITAMIN-D
ON INVERTED PALINDROMES
OFF domain
Carr RW;Gregory JE;Proske U;
1998
1043
Summation of responses of cat muscle spindles to
combined static and dynamic fusimotor stimulation
OFF domain
Carr WES;Derby CD;
1986
209
BEHAVIORAL CHEMOATTRACTANTS FOR THE SHRIMP,
PALAEMONETES-PUGIO - IDENTIFICATION OF ACTIVE
COMPONENTS IN FOOD EXTRACTS AND EVIDENCE OF
SYNERGISTIC MIXTURE INTERACTIONS
ON trivial
Carrasco F;Ruz M;Rojas P;Csendes
A;Rebolledo A;Codoceo J;Inostroza J;Basfi-fer
K;Papapietro K;Rojas J;Pizarro F;Olivares M;
2009
560
Changes in Bone Mineral Density, Body Composition and
Adiponectin Levels in Morbidly Obese Patients after
Bariatric Surgery
OFF domain
Carvounis G;Carvounis CP;Arbeit LA;
1985
185
INDEPENDENT ACTION OF PROSTAGLANDINS AND
KININS ON VASOPRESSIN-STIMULATED WATER-FLOW
OFF domain
Casner PR;Goldman HW;Lehr D;
1976
46
IMPORTANCE OF ANGIOTENSIN-II FORMATION IN CNS
IN ANGIOTENSIN-I - ELICITED THIRST
OFF domain
Castanedaagullo M;Delcastillo LM;
1962
5
ON DIELECTRIC CONSTANT AND ENZYMATIC KINETICS
.4. DIPOLAR IONS - ESTER HYDROLYSIS BY TRYPSIN
AND ALPHA CHYMOTRYPSIN IN GLYCINE SOLUTIONS
OFF domain
Castelnovo L;Dosquet C;Gaudric A;Sahel
J;Hicks D;
2000
775
Human platelet suspension stimulates porcine retinal glial
proliferation and migration in vitro
OFF domain
Castle JC;Zhang CL;Shah JK;Kulkarni
AV;Kalsotra A;Cooper TA;Johnson JM;
2008
406
Expression of 24,426 human alternative splicing events
and predicted cis regulation in 48 tissues and cell lines
OFF domain
APPENDIX A
Authors
Page 8 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Cedergreen N;Abbaspoor M;Sorensen
H;Streibig JC;
2007
550
Is mixture toxicity measured on a biomarker indicative of
what happens on a population level? A study with Lemna
minor
ON experimental
study
Cedergreen N;Christensen AM;Kamper
A;Kudsk P;Mathiassen SK;Streibig
JC;Sorensen H;
2008
553
A review of independent action compared to concentration
addition as reference models for mixtures of compounds
with different molecular target sites
ON experimental
study
Cedergreen N;Kamper A;Streibig JC;
2006
557
Is prochloraz a potent synergist across aquatic species? A
study on bacteria, daphnia, algae and higher plants
ON experimental
study
Cedergreen N;Kudsk P;Mathiassen
SK;Sorensen H;Streibig JC;
2007
470
Reproducibility of binary-mixture toxicity studies
ON experimental
study
Cedergreen N;Kudsk P;Mathiassen
SK;Streibig JC;
2007
807
Combination effects of herbicides on plants and algae: do
species and test systems matter?
ON experimental
study
Cedergreen N;Streibig JC;
2005
1034
Can the choice of endpoint lead to contradictory results of
mixture-toxicity experiments?
ON review
Cerra A;Mansfield KJ;Chamberlain CG;
2003
778
Exacerbation of TGF-beta-induced cataract by FGF-2 in
cultured rat lenses
OFF domain
Chaban VV;McRoberts JA;Ennes HS;Mayer EA;
2001
707
Nitric oxide synthase inhibitors enhance mechanosensitive
Ca2+ influx in cultured dorsal root ganglion neurons
OFF domain
Chabaud M;Durand JM;Buchs N;Fossiez
F;Page G;Frappart L;Miossec P;
1999
716
Human interleukin-17 - A T cell-derived proinflammatory
cytokine produced by the rheumatoid synovium
OFF domain
Chabchoub-Ellouze S;Hamdaoui MH;
2006
734
Comparative effects of the addition of mutton or beef to a
bean seed ragout on iron status in growing rats
OFF domain
Chakraborti C;Egan J;
2010
457
The lesser of two adverse reactions
OFF domain
Chamiot-Clerc P;Choukri N;Legrand M;Safar
ME;Droy-Lefaix MT;Renaud JF;
1999
723
Endothelium-dependent and independent properties of
cicletanine in Wistar rat aorta under normoxic and hypoxic
conditions
OFF domain
Charlesworth D;
1988
254
EVOLUTION OF HOMOMORPHIC SPOROPHYTIC SELFINCOMPATIBILITY
OFF domain
Charlton MR;Adey DB;Nair KS;
1996
513
Evidence for a catabolic role of glucagon during an amino
acid load
OFF domain
Chatterjee H;
2005
830
Studies on the synergistic response of some commercial
biopesticides with botanicals, growth regulator and
conventional organophosphate against neonate larvae of
Phthorimaea operculella (Zeller)
ON experimental
study
Chen CY;Chen SL;Christensen ER;
2005
369
Individual and combined toxicity of nitriles and aldehydes
to Raphidocelis subcapitata
ON experimental
study
Chen CY;Lu CL;
2002
1047
An analysis of the combined effects of organic toxicants
ON experimental
study
Chen H;Yao J;Wang F;Zhou Y;Chen
K;Zhuang R;Choi MMF;Zaray G;
2010
410
Toxicity of three phenolic compounds and their mixtures
on the gram-positive bacteria Bacillus subtilis in the
aquatic environment
ON experimental
study
Chen JL;Huisinga KL;Viering MM;Ou SA;Wu
CT;Geyer PK;
2002
637
Enhancer action in trans is permitted throughout the
Drosophila genome
OFF domain
Chen X;Li Z;Li J;
2002
726
Anti-inflammatory effect of cerivastatin in vascular injury
independent of serum cholesterol and blood pressure
lowering effects in mouse model
OFF domain
Cheng K;Raufman JP;
2005
340
Autocrine release of HB-EGF mediates bile acid-induced
transactivation of colon cancer cell epidermal growth
factor receptors (EGFR)
OFF domain
Chernichenko IA;
1989
282
FEATURES OF THE CARCINOGENIC EFFECT OF BENZO A
PYRENE ENTERING THE BODY VIA DIFFERENT ROUTES
AND THE PROBLEM OF ITS STANDARDIZATION IN THE
ENVIRONMENT
OFF domain
Chesnoy-Marchais D;
2005
832
The estrogen receptor modulator tamoxifen enhances
spontaneous glycinergic synaptic inhibition of hypoglossal
motoneurons
OFF domain
APPENDIX A
Authors
Page 9 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Chevre N;Loeppe C;Singer H;Stamm
C;Fenner K;Escher BI;
2006
688
Including mixtures in the determination of water quality
criteria for herbicides in surface water
ON experimental
study
Chiou PWS;Chen C;Yu B;
2000
854
Effects of Aspergillus oryzae fermentation extract on in
situ degradation of feedstuffs
OFF domain
Chiu AT;Roscoe WA;McCall DE;Timmermans
PB;
1991
1003
Angiotensin II-1 receptors mediate both vasoconstrictor
and hypertrophic responses in rat aortic smooth muscle
cells
OFF domain
Chouaib S;Bensussan A;Termijtelen
AM;Andreeff M;Marchiolfournigault C;Fradelizi
D;Dupont B;
1988
255
ALLOGENEIC T-CELL ACTIVATION TRIGGERING BY MHC
CLASS-I ANTIGENS
OFF domain
Christensen AM;Ingerslev F;Baun A;
2006
689
Ecotoxicity of mixtures of antibiotics used in aquacultures
ON experimental
study
Christensen S;
1983
153
EFFECTS OF LITHIUM ON WATER-INTAKE AND RENAL
CONCENTRATING ABILITY IN RATS WITH VASOPRESSINDEFICIENT DIABETES-INSIPIDUS (BRATTLEBORO
STRAIN)
OFF domain
Christian ML;Owens JC;Lewis BE;Davis FM;
1986
210
INTERACTION STUDIES OF METHYL PARATHION,
ON trivial
CARBOFURAN, PERMETHRIN, AND PIPERONYL BUTOXIDE
APPLIED TO SOUTHWESTERN CORN-BORER LARVAE
(LEPIDOPTERA, PYRALIDAE)
Christiansen S;Scholze M;Dalgaard
M;Vinggaard AM;Axelstad M;Kortenkamp
A;Hass U;
2009
451
Synergistic Disruption of External Male Sex Organ
Development by a Mixture of Four Antiandrogens
ON experimental
study
Chu YP;Cheng YC;Yang CC;Chang LS;
2005
596
The structural events associated with the binding of
divalent cations to beta-bungarotoxin
OFF domain
Chueden HG;Weinmann HM;
1980
521
THE BIPHASIC RESPONSE IN EVOKED RESPONSE
AUDIOMETRY
OFF domain
Civillico EF;Contreras D;
2006
500
Integration of evoked responses in supragranular cortex
studied with optical recordings in vivo
OFF domain
Clark A;Weymann A;Hartman E;Turmelle
YP;Hourcade D;Rudnick D;
2007
341
Evidence for alternative- and classical-pathway
independent action of complement factor C3 during liver
regeneration
OFF domain
Clarke B;
1987
656
Arthur Wigan and The Duality of the Mind
OFF domain
Clayton RD;Duffy SR;Wilkinson N;Garry
R;Jackson AM;
2004
770
Anti-proliferative effect of mycobacteria, IFN-gamma and
TNF-alpha on primary cultures derived from endometrial
stroma: Possible relevance to endometriosis?
OFF domain
Cleuvers M;
2003
1064
Aquatic ecotoxicity of pharmaceuticals including the
assessment of combination effects
ON experimental
study
Cleuvers M;Altenburger R;Ratte HT;
2002
1012
Combination effect of light and toxicity in algal tests
ON experimental
study
Coates TW;Langley PA;
1982
132
THE CAUSES OF MATING ABSTENTION IN MALE TSETSE
FLIES GLOSSINA-MORSITANS
OFF domain
Cometto-Muniz JE;Cain WS;Abraham MH;
2005
960
Odor detection of single chemicals and binary mixtures
ON experimental
study
Coors A;De Meester L;
2008
361
Synergistic, antagonistic and additive effects of multiple
stressors: predation threat, parasitism and pesticide
exposure in Daphnia magna
ON experimental
study
Corbett DA;Goldberg MT;Swamy VC;Triggle
CR;Triggle DJ;
1980
87
REACTIVITY OF VASA DEFERENTIA FROM
SPONTANEOUSLY HYPERTENSIVE AND NORMOTENSIVE
WISTAR RATS
OFF domain
Cosgrove L;McGeechan PL;Handley
PS;Robson GD;
2010
1057
Effect of Biostimulation and Bioaugmentation on
Degradation of Polyurethane Buried in Soil
OFF domain
Cosiski Marana HR;Santana da Silva J;Moreira
de Andrade J;
2000
721
NK cell activity in the presence of IL-12 is a prognostic
assay to neoadjuvant chemotherapy in cervical cancer
OFF domain
Cossins AR;Kilbey RV;
1991
319
ADRENERGIC RESPONSES AND THE ROOT EFFECT IN
ERYTHROCYTES OF FRESH-WATER FISH
OFF domain
APPENDIX A
Authors
Page 10 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Cote GP;Smillie LB;
1981
107
THE EFFECTS OF PLATELET TROPOMYOSIN ON THE
ATPASE ACTIVITIES OF MUSCLE ACTOMYOSIN
SUBFRAGMENT-1 IN THE ABSENCE AND PRESENCE OF
TROPONIN, ITS COMPONENTS, AND CALMODULIN
OFF domain
Courtiss EH;Ransil BJ;Russo J;
1995
583
THE EFFECTS OF HYALURONIDASE ON LOCALANESTHESIA - A PROSPECTIVE, RANDOMIZED,
CONTROLLED, DOUBLE-BLIND-STUDY
OFF domain
Cox C;
1992
783
A GLM APPROACH TO QUANTAL RESPONSE MODELS FOR
MIXTURES
ON method dev.
Craven PA;Briggs R;Derubertis FR;
1980
88
CALCIUM-DEPENDENT ACTION OF OSMOLALITY ON
ADENOSINE-3',5'-MONOPHOSPHATE ACCUMULATION IN
RAT RENAL INNER MEDULLA - EVIDENCE FOR A
RELATIONSHIP TO CALCIUM-RESPONSIVE
ARACHIDONATE RELEASE AND PROSTAGLANDIN
SYNTHESIS
OFF domain
Crow AR;Song S;Semple JW;Freedman
J;Lazarus AH;
2001
413
IVIg inhibits reticuloendothelial system function and
ameliorates murine passive-immune thrombocytopenia
independent of anti-idiotype reactivity
OFF domain
Crowder M;
1997
939
A test for independence of competing risks with discrete
failure times
OFF domain
Currie SM;
1999
725
Aspects of the preparation of student midwives for
autonomous practice
OFF domain
Cuthbert AW;Painter E;
1968
9
INDEPENDENT ACTION OF ANTIDIURETIC HORMONE
THEOPHYLLINE AND CYCLIC 3',5'-ADENOSINE
MONOPHOSPHATE ON CELL MEMBRANE PERMEABILITY
IN FROG SKIN
OFF domain
Cutilli T;Papola F;Di Emidio P;Corbacelli A;
1997
744
p53 mutation and chemoresistance in oral-maxillofacial
squamous cell carcinoma. Role of p53 in the cell cycle
control and in the modulating action of chemotherapeutic
agents
OFF domain
Dahl CD;Logothetis NK;Kayser C;
2009
703
Spatial Organization of Multisensory Responses in
Temporal Association Cortex
OFF domain
Dai W;Kamei H;Zhao Y;Ding J;Du Z;Duan CM;
2010
632
Duplicated zebrafish insulin-like growth factor binding
protein-5 genes with split functional domains: evidence for
evolutionarily conserved IGF binding, nuclear localization,
and transactivation activity
OFF domain
Damle NK;Leytze G;Klussman K;Ledbetter JA;
1993
822
ACTIVATION WITH SUPERANTIGENS INDUCES
OFF domain
PROGRAMMED DEATH IN ANTIGEN-PRIMED CD4+ CLASSII+ MAJOR HISTOCOMPATIBILITY COMPLEX TLYMPHOCYTES VIA A CD11A/CD18-DEPENDENT
MECHANISM
Daniel PC;Derby CD;
1987
229
MIXTURE INTERACTION ANALYSIS - A POLYNOMIAL
RESPONSE SUMMATION MODEL WHICH INCORPORATES
THE BEIDLER EQUATION
ON trivial
Dardenne F;Nobels I;De Coen W;Blust R;
2008
433
Mixture toxicity and gene inductions: Can we predict the
outcome?
ON experimental
study
Davidovich L;Zhu SY;Khoury AZ;Su C;
1992
863
SUITABILITY OF THE MASTER-EQUATION APPROACH
FOR MICROMASERS WITH NON-POISSONIAN PUMPING
OFF domain
Dawson DA;
1991
964
JOINT ACTION OF TRANSFER-RETINOIC AND VALPROIC
ACIDS ON XENOPUS EMBRYO DEVELOPMENT
ON experimental
study
Dawson DA;
1994
963
JOINT ACTIONS OF CARBOXYLIC-ACID BINARYMIXTURES ON XENOPUS EMBRYO DEVELOPMENT COMPARISON OF JOINT ACTIONS FOR MALFORMATION
TYPES
ON experimental
study
Dawson DA;Wilke TS;
1991
320
INITIAL EVALUATION OF DEVELOPMENTAL
MALFORMATION AS AN END-POINT IN MIXTURE
TOXICITY HAZARD ASSESSMENT FOR AQUATIC
VERTEBRATES
ON trivial
APPENDIX A
Authors
Page 11 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Dawson DA;Wilke TS;
1991
965
EVALUATION OF THE FROG EMBRYO TERATOGENESIS
ASSAY - XENOPUS (FETAX) AS A MODEL SYSTEM FOR
MIXTURE TOXICITY HAZARD ASSESSMENT
ON experimental
study
Dawson DA;Wilke TS;
1992
322
JOINT ACTIONS OF DEVELOPMENTAL TOXICANTS IN
XENOPUS EMBRYOS - BINARY-MIXTURES OF DNASYNTHESIS INHIBITORS
ON trivial
de Alwis SP;
2006
374
Transitions between flux vacua
OFF domain
De Zwart D;Posthuma L;
2005
679
Complex mixture toxicity for single and multiple species:
Proposed methodologies
ON review
Decking UKM;Juengling E;Kammermeier H;
1988
256
INTERSTITIAL TRANSUDATE CONCENTRATION OF
ADENOSINE AND INOSINE IN RAT AND GUINEA-PIG
HEARTS
OFF domain
Delhanty PJD;Sun Y;Visser JA;van Kerkwijk
A;Huisman M;van Ijcken WFJ;Swagemakers
S;Smith RG;Themmen APN;van der Lely A;
2010
1038
Unacylated ghrelin rapidly modulates lipogenic and insulin
signaling pathway gene expression in metabolically active
tissues of GHSR deleted mice
OFF domain
delRio G;Morett E;Soberon X;
1997
937
Did cyclodextrin glycosyltransferases evolve from alphaamylases?
OFF domain
Delyra JL;Dewitt B;Foong SK;Gallivan
TE;Harrington R;Kapulkin A;Myers
E;Polchinski J;
1992
380
THE QUANTIZED O(1,2)/O(2)XZ2 SIGMA MODEL HAS NO
CONTINUUM-LIMIT IN 4 DIMENSIONS .2. LATTICE
SIMULATION
OFF domain
Demarch BGE;
1987
230
SIMPLE SIMILAR ACTION AND INDEPENDENT JOINT
ACTION - 2 SIMILAR MODELS FOR THE JOINT EFFECTS
OF TOXICANTS APPLIED AS MIXTURES
ON trivial
Deng Fc;Liu Ss;Liu Hl;Mo Ly;
2007
1091
Toxicities of selected heavy metal compounds and their
mixtures to photobacteria(Vibrio-qinghaiensis sp.-Q67)
ON experimental
study
Derubertis FR;Craven PA;
1978
63
ALTERATIONS IN RAT RENAL CORTICAL AND
MEDULLARY GUANOSINE 3' 5'-MONOPHOSPHATE
ACCUMULATION BY OXYGEN-DEPENDENT AND CALCIUMDEPENDENT AND INDEPENDENT MECHANISMS EVIDENCE FOR A CALCIUM-INDEPENDENT ACTION OF
OXYGEN IN RENAL INNER MEDULLA
OFF domain
Derubertis FR;Craven PA;
1979
76
PROPERTIES OF SOLUBLE CYCLIC AMP-DEPENDENT
PROTEIN-KINASE ACTIVITY OF RENAL INNER MEDULLA
OFF domain
Deser S;Jackiw R;
1999
1119
Higher derivative Chern-Simons extensions
OFF domain
Deverina IS;
1980
951
Mechanism of inhibition by chlorgyline and deprenyl of
tyramine deamination by mitochondrial monoamine
oxidase of rat liver
OFF domain
Diaz A;Chepenik KP;Korn JH;Reginato
AM;Jimenez SA;
1998
611
Differential regulation of cyclooxygenases 1 and 2 by
interleukin-1 beta, tumor necrosis factor-alpha, and
transforming growth factor-beta 1 in human lung
fibroblasts
OFF domain
Dilbeck GA;Field L;Gallo AA;Gargiulo RJ;
1978
64
BIOLOGICALLY ORIENTED ORGANIC SULFUR CHEMISTRY
.19. SYNTHESIS AND PROPERTIES OF '2-AMINO-5MERCAPTO-5-METHYLHEXANOIC ACID, A
BISHOMOLOGUE OF PENICILLAMINE - USE OF BORONTRIFLUORIDE ETHERATE FOR CATALYZING
MARKOVNIKOV ADDITION OF A THIOL TO AN OLEFIN
OFF domain
DiMario MJ;Boardman JT;Sauser BJ;
2009
927
System of Systems Collaborative Formation
OFF domain
Dimatteo MR;Reiter RC;Gambone JC;
1994
924
ENHANCING MEDICATION ADHERENCE THROUGH
COMMUNICATION AND INFORMED COLLABORATIVE
CHOICE
OFF domain
Dincer B;Erbas D;
2010
1051
Description of Communication Breakdown Repair
Strategies Produced By Nonverbal Students with
Developmental Disabilities
OFF domain
Ding W;Li CX;Hu TH;Graves-Deal R;Fotia
AB;Weissman AM;Coffey RJ;
2008
700
EGF receptor-independent action of TGF-alpha protects
Naked2 from AO7-mediated ubiquitylation and
proteasomal degradation
OFF domain
APPENDIX A
Authors
Page 12 of 66
PubYear
ID
CFT/EFSA/PPR/2010/02
Title
Relevance
Dittman L;
2000
1029
Overfiling: Policy arguments in support of the gorilla in the
closet
OFF domain
Dixon RC;Hamilton PB;
1981
108
EFFECT OF FEED INGREDIENTS ON THE ANTIFUNGAL
ACTIVITY OF PROPIONIC-ACID
OFF domain
Doering WV;Wang YH;
1999
837
CryptoCope rearrangement of 1,3-dicyano-5-phenyl-4,4d(2)-hexa-2,5-diene. Chameleonic or centauric?
OFF domain
Doermer P;Boehmer RM;
1984
929
CELL CYCLE-DEPENDENT AND CELL CYCLEINDEPENDENT DAMAGE TO RAT HEMOPOIESIS BY
HYDROXYUREA
OFF domain
Doganay G;Khodr B;Georgiou G;Khalil Z;
2006
858
Pharmacological manipulation of the vasoconstrictive
effects of amyloid beta peptides by donepezil and
rivastigmine
OFF domain
Dolan TT;Young AS;Losos GJ;McMilian
I;Minder CE;Soulsby K;
1984
163
DOSE DEPENDENT RESPONSES OF CATTLE TO
THEILERIA-PARVA STABILATE
OFF domain
Dominguez-Salazar E;Shetty S;Rissman EF;
2006
1017
Rapid neural fos responses to oestradiol in oestrogen
receptor alpha beta double knockout mice
OFF domain
Doppler W;Windegger M;Soratroi C;Tomasi
J;Lechner J;Rusconi S;Cato ACB;Almlof
T;Liden J;Okret S;Gustafsson JA;Richard-Foy
H;Starr DB;Klocker H;Edwards D;Geymayer S;
2001
843
Expression level-dependent contribution of glucocorticoid
receptor domains for functional interaction with STAT5
OFF domain
Dormans TPJ;Gerlag PGG;Russel FGM;Smits
P;
1998
813
Combination diuretic therapy in severe congestive heart
failure
OFF domain
Dormer P;Bohmer RM;
1984
164
CELL-CYCLE-DEPENDENT AND CELL-CYCLEINDEPENDENT DAMAGE TO RAT HEMATOPOIESIS BY
HYDROXYUREA
OFF domain
dos Santos NA;Simas MLD;Nogueira R;
2007
855
Visual contrast perception in human beings:
Psychophysical evidence for high angular frequency
channels
OFF domain
Doty P;Dykstra LA;Picker MJ;
1992
907
DELTA-9-TETRAHYDROCANNABINOL INTERACTIONS
WITH PHENCYCLIDINE AND ETHANOL - EFFECTS ON
ACCURACY AND RATE OF RESPONDING
OFF domain
Doughty MJ;Diehn B;
1984
165
ANION SENSITIVITY OF MOTILITY AND STEP-DOWN
PHOTOPHOBIC RESPONSES OF EUGLENA-GRACILIS
OFF domain
Draper MJ;
1982
133
A CHILDS INDEPENDENT ACTION FOR LOSS OF
CONSORTIUM - A CHANGE IN THE IOWA TORT SCHEME
OFF domain
Drescher K;Boedeker W;
1995
865
ASSESSMENT OF THE COMBINED EFFECTS OF
ON experimental
SUBSTANCES - THE RELATIONSHIP BETWEEN
study
CONCENTRATION ADDITION AND INDEPENDENT ACTION
Drewes CD;Pax RA;
1974
28
NEUROMUSCULAR PHYSIOLOGY OF LONGITUDINAL
MUSCLE OF EARTHWORM, LUMBRICUS-TERRESTRIS .2.
PATTERNS OF INNERVATION
OFF domain
Droese S;Brandt U;Hanley PJ;
2006
394
K+-independent actions of diazoxide question the role of
inner membrane K-ATP channels in mitochondrial
cytoprotective signaling
OFF domain
Drucker EG;Lauder GV;
2000
332
Function of the teleost dorsal fin: experimental analysis of
wake forces in sunfish
OFF domain
Drummherrel H;Mohr H;
1991
825
INVOLVEMENT OF PHYTOCHROME IN LIGHT CONTROL
OF STEM ELONGATION IN CUCUMBER (CUCUMISSATIVUS L) SEEDLINGS
OFF domain
Drumond YA;Couto AS;Moraessantos
T;Almeida AP;Freiremaia L;
1995
912
EFFECTS OF TOXIN TS-GAMMA AND TITYUSTOXIN
PURIFIED FROM TITYUS-SERRULATUS SCORPIONVENOM ON ISOLATED RAT ATRIA
OFF domain
Duan C;
2001
334
Extracellular and nuclear actions of insulin-like growth
factor binding proteins
OFF domain
Duane WC;Xiong W;Lofgren J;
2008
1089
Transactivation of the human apical sodium-dependent
bile acid transporter gene by human serum
OFF domain
APPENDIX A
Authors
Page 13 of 66
PubYear
ID
CFT/EFSA/PPR/2010/02
Title
Relevance
Duane WC;Xiong W;Wolvers J;
2007
1088
Effects of bile acids on expression of the human apical
sodium dependent bile acid transporter gene
OFF domain
Duran-Sandoval D;Mautino G;Martin
GV;Percevault F;Barbier O;Fruchart
JC;Kuipers F;Staels B;
2004
421
Glucose regulates the expression of the farnesoid X
receptor in liver
OFF domain
Durrant JD;
1987
231
AUDITORY-EVOKED POTENTIAL TO PATTERN-REVERSAL
STIMULATION
OFF domain
Dyer S;Warne MS;Meyer JS;Leslie HA;Escher
BI;
2011
434
Tissue Residue Approach for Chemical Mixtures
ON review
Earley CJ;Leonard BE;
1978
65
BEHAVIORAL-STUDIES ON EFFECTS OF D-AMPHETAMINE
AND ESTRADIOL BENZOATE ALONE AND IN
COMBINATION
OFF domain
Ebling FJ;Ebling E;Randall V;Skinner J;
1975
36
EFFECTS OF HYPOPHYSECTOMY AND OF BOVINE
GROWTH-HORMONE ON RESPONSES TO TESTOSTERONE
OF PROSTATE, PREPUTIAL, HARDERIAN AND LACRIMAL
GLANDS AND OF BROWN ADIPOSE-TISSUE IN RAT
OFF domain
Echeveste P;Dachs J;Berrojalbiz N;Agusti S;
2010
667
Decrease in the abundance and viability of oceanic
phytoplankton due to trace levels of complex mixtures of
organic pollutants
ON experimental
study
Edelstein M;Valeriote F;Vietti T;
1978
66
INFLUENCE OF CONTINUOUS INFUSION OF CYTOSINEARABINOSIDE ON SEQUENCING WITH DAUNORUBICIN
IN L1210 LEUKEMIA
OFF domain
Eden OB;Sills JA;Brown JK;
1977
52
HYPERTENSION IN ACUTE NEUROLOGICAL DISEASES OF
CHILDHOOD
OFF domain
Edwards TL;Singleton VL;Boulton R;
1985
186
FORMATION OF ETHYL-ESTERS OF TARTARIC ACID
DURING WINE AGING - CHEMICAL AND SENSORY
EFFECTS
OFF domain
Eggleston P;Kearsey MJ;
1980
89
HYBRID DYSGENESIS IN DROSOPHILA - CORRELATION
BETWEEN DYSGENIC TRAITS
OFF domain
Ellakwah F;Meuser F;Gawad AA;Wohlgemuth
R;Darwish A;
1991
1094
EFFICIENCY OF PHOSPHINE ALONE AND IN MIXTURES
WITH CARBON-DIOXIDE AGAINST ANGOUMOIS GRAIN
MOTH SITOTROGA-CEREALELLA (OLIVIER),
(GELECHIIDAE, LEPIDOPTERA)
OFF domain
Ellger B;Richir MC;van Leeuwen
PAM;Debaveye Y;Langouche L;Vanhorebeek
I;Teerlink T;Van den Berghe G;
2008
606
Glycemic control modulates arginine and asymmetricaldimethylarginine levels during critical illness by preserving
dimethylarginine-dimethylaminohydrolase activity
OFF domain
Ellies LG;Heersche JNM;Vadas P;Pruzanski
W;Stefanski E;Aubin JE;
1991
1109
INTERLEUKIN-1-ALPHA STIMULATES THE RELEASE OF
PROSTAGLANDIN-E2 AND PHOSPHOLIPASE-A2 FROM
FETAL-RAT CALVARIAL CELLS-INVITRO - RELATIONSHIP
TO BONE NODULE FORMATION
OFF domain
Ellis AG;Adam WR;Martin TJ;
1990
300
COMPARISON OF THE EFFECTS OF PARATHYROIDHORMONE (PTH) AND RECOMBINANT PTH-RELATED
PROTEIN ON BICARBONATE EXCRETION BY THE
ISOLATED PERFUSED RAT-KIDNEY
OFF domain
Ellis E;Axelson M;Abrahamsson A;Eggertsen
G;Thorne A;Nowak G;Ericzon BG;Bjorkhem
I;Einarsson C;
2003
769
Feedback regulation of bile acid synthesis in primary
human hepatocytes: Evidence that CDCA is the strongest
inhibitor
OFF domain
Ely JA;Hunyady L;Baukal AJ;Catt KJ;
1990
301
INOSITOL 1,3,4,5-TETRAKISPHOSPHATE STIMULATES
CALCIUM RELEASE FROM BOVINE ADRENAL
MICROSOMES BY A MECHANISM INDEPENDENT OF THE
INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR
OFF domain
Emel'ianova TG;Goriacheva NN;Guzavatykh
LS;Andreeva LA;Alfeeva LI;Miasoedov NF;
2004
1118
Thermoregulatory activity of dermorphin fragments
OFF domain
Emery KM;Scherm H;Savelle AT;
2002
686
Assessment of interactions between components of
fungicide mixtures against Monilinia fructicola
ON experimental
study
APPENDIX A
Authors
Page 14 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Endoh M;
1989
283
REGULATION OF FORCE AND INTRACELLULAR CALCIUM
TRANSIENTS BY CYCLIC-AMP GENERATED BY
FORSKOLIN, MDL-17,043 AND ISOPRENALINE, AND ITS
MODULATION BY MUSCARINIC RECEPTOR AGENTS - A
NOVEL MECHANISM FOR ACCENTUATED ANTAGONISM
OFF domain
Engels WR;
1979
77
HYBRID DYSGENESIS IN DROSOPHILA-MELANOGASTER RULES OF INHERITANCE OF FEMALE STERILITY
OFF domain
Enwere NJ;Hung YC;Ngoddy PO;
1990
976
TEXTURE AND MICROSTRUCTURE OF AFRICAN YAM
BEAN (SPHENOSTYLIS-STERNOCARPA) PRODUCTS
OFF domain
Erkelens CJ;Regan D;
1986
211
HUMAN OCULAR VERGENCE MOVEMENTS INDUCED BY
CHANGING SIZE AND DISPARITY
OFF domain
Erlenkaemper B;Bugiel C;Brecher C;Werth
S;Eisentraeger A;
2008
568
Environmental assessment of ester-based lubricants after
application
ON experimental
study
Esashi Y;Fuwa N;Kurota A;Oota H;Abe M;
1987
232
INTERRELATION BETWEEN ETHYLENE AND CARBONDIOXIDE IN RELATION TO RESPIRATION AND
ADENYLATE CONTENT IN THE PRE-GERMINATION
PERIOD OF COCKLEBUR SEEDS
OFF domain
Escher BI;Bramaz N;Eggen RIL;Richter M;
2005
417
In vitro assessment of modes of toxic action of
pharmaceuticals in aquatic life
ON experimental
study
Escher BI;Bramaz N;Lienert J;Neuwoehner
J;Straub JO;
2010
831
Mixture toxicity of the antiviral drug Tamiflu (R)
(oseltamivir ethylester) and its active metabolite
oseltamivir acid
ON experimental
study
Escher BI;Bramaz N;Maurer M;Richter
M;Sutter D;von Kanel C;Zschokke M;
2005
392
Screening test battery for pharmaceuticals in urine and
wastewater
ON trivial
Eshel Y;Salomon Y;
1994
979
CALMODULIN-BINDING PEPTIDES INTERFERE WITH
MELANOCYTE-STIMULATING HORMONE-RECEPTOR
ACTIVITY AND STIMULATE ADENOSINE 3,5MONOPHOSPHAT PRODUCTION IN M2R MOUSE
MELANOMA-CELLS
OFF domain
Evans MJ;Mulligan RS;Livesey JH;Donald RA;
1996
747
The integrative control of adrenocorticotrophin secretion:
A critical role for corticotrophin-releasing hormone
OFF domain
F. R. Cassee;J. P. Groten;P. J. van
Bladeren;V. J. Feron
1998
1132
Toxicological evaluation and risk assessment of chemical
mixtures
ON review
Fantino E;Savastano HI;
1996
1075
Humans' responses to novel stimulus compounds and the
effects of training
OFF domain
Farquhar M;Soomets U;Bates RL;Martin
A;Langel U;Howl J;
2002
487
Novel mastoparan analogs induce differential secretion
from mast cells
OFF domain
Farthing MJG;Mattei AM;Edwards
CRW;Dawson AM;
1982
134
RELATIONSHIP BETWEEN PLASMA TESTOSTERONE AND
DIHYDROTESTOSTERONE CONCENTRATIONS AND MALE
FACIAL HAIR-GROWTH
OFF domain
Faust M;Altenburger R;Backhaus T;Blanck
H;Boedeker W;Gramatica P;Hamer V;Scholze
M;Vighi M;Grimme LH;
2001
574
Predicting the joint algal toxicity of multi-component striazine mixtures at low-effect concentrations of individual
toxicants
ON experimental
study
Faust M;Altenburger R;Backhaus T;Blanck
H;Boedeker W;Gramatica P;Hamer V;Scholze
M;Vighi M;Grimme LH;
2003
548
Joint algal toxicity of 16 dissimilarly acting chemicals is
predictable by the concept of independent action
ON experimental
study
Faust M;Altenburger R;Backhaus T;Bodeker
W;Scholze M;Grimme LH;
2000
509
Predictive assessment of the aquatic toxicity of multiple
chemical mixtures
ON experimental
study
Feddersen CO;Chang S;Czartalomna J;Voelkel
NF;
1990
302
ARACHIDONIC-ACID CAUSES CYCLOOXYGENASEDEPENDENT AND CYCLOOXYGENASE-INDEPENDENT
PULMONARY VASODILATION
OFF domain
Federici M;Natoli S;Bernardi G;Mercuri NB;
2002
928
Dopamine selectively reduces GABA(B) transmission onto
dopaminergic neurones by an unconventional presynaptic
action
OFF domain
Federle TW;Vestal JR;
1980
90
LIGNOCELLULOSE MINERALIZATION BY ARCTIC LAKESEDIMENTS IN RESPONSE TO NUTRIENT MANIPULATION
OFF domain
APPENDIX A
Authors
Page 15 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Fedorova TN;
1982
135
PRIMARY OCTOPLOID TRITICALE, MEIOSIS STABILITY
AND SELECTION
OFF domain
Feofilova EP;Gryaznova MV;Sadovova NV;
1989
902
THE ABILITY OF MANNITOL AND TREHALOSE TO
INCREASE THE RESISTANCE OF MYCELIAL FUNGI TO
TEMPERATURE CHANGES
OFF domain
Ferguson DO;Rice MC;Rendi MH;Kotani
H;Kmiec EB;Holloman WK;
1997
377
Interaction between Ustilago maydis REC2 and RAD51
genes in DNA repair and mitotic recombination
OFF domain
Ferreira ALG;Loureiro S;Soares A;
2008
534
Toxicity prediction of binary combinations of cadmium,
carbendazim and low dissolved oxygen on Daphnia magna
ON experimental
study
Ferreira ALG;Serra P;Soares A;Loureiro S;
2010
565
The influence of natural stressors on the toxicity of nickel
to Daphnia magna
ON experimental
study
Fine JS;Byrnes HD;Zavodny PJ;Hipkin RW;
2001
958
Evaluation of signal transduction pathways in
chemoattractant-induced human monocyte chemotaxis
OFF domain
Finkelman FD;Svetic A;Gresser I;Snapper
C;Holmes J;Trotta PP;Katona IM;Gause WC;
1991
431
REGULATION BY INTERFERON-ALPHA OF
IMMUNOGLOBULIN ISOTYPE SELECTION AND
LYMPHOKINE PRODUCTION IN MICE
OFF domain
Fischer BB;Rufenacht K;Dannenhauer
K;Wiesendanger M;Eggen RIL;
2010
544
MULTIPLE STRESSOR EFFECTS OF HIGH LIGHT
IRRADIANCE AND PHOTOSYNTHETIC HERBICIDES ON
GROWTH AND SURVIVAL OF THE GREEN ALGA
CHLAMYDOMONAS REINHARDTII
OFF domain
Fishwild DM;Benike CJ;Engleman EG;
1988
257
ACTIVATION OF HLA-RESTRICTED EBV-SPECIFIC CYTOTOXIC T-CELLS DOES NOT REQUIRE CD4+ (HELPER) TCELLS OR EXOGENOUS CYTOKINES
OFF domain
Flamigni F;Guarnieri C;Caldarera CM;
1988
258
RAT-LIVER CYTOSOL CONTAINS NADPH-DEPENDENT
AND GSH-DEPENDENT FACTORS ABLE TO RESTORE
ORNITHINE DECARBOXYLASE INACTIVATED BY
REMOVAL OF THIOL REDUCING AGENTS
OFF domain
Flippin JL;Hedge JM;DeVito MJ;LeBlanc
GA;Crofton KM;
2009
1061
Predictive Modeling of a Mixture of Thyroid Hormone
ON experimental
Disrupting Chemicals That Affect Production and Clearance study
of Thyroxine
Flores JA;Leong DA;Veldhuis JD;
1992
1106
IS THE CALCIUM SIGNAL INDUCED BY FOLLICLESTIMULATING-HORMONE IN SWINE GRANULOSA-CELLS
MEDIATED BY ADENOSINE CYCLIC 3',5'MONOPHOSPHATE-DEPENDENT PROTEIN-KINASE
OFF domain
Florijn WJ;Deboer T;Tonnaer J;Versteeg DHG;
1992
398
CHARACTERIZATION OF THE INHIBITORY EFFECT OF
ADRENOCORTICOTROPIN MELANOCYTE-STIMULATING
HORMONE-LIKE PEPTIDES ON THE BINDING OF
DOPAMINE RECEPTOR LIGANDS TO THE DOPAMINE-D2
RECEPTOR INVITRO
OFF domain
Foltin RW;Fischman MW;
1992
642
THE CARDIOVASCULAR AND SUBJECTIVE EFFECTS OF
INTRAVENOUS COCAINE AND MORPHINE
COMBINATIONS IN HUMANS
OFF domain
Fontan E;Fauve RM;
1986
212
INCREASED PHAGOCYTIC-ACTIVITY IN MICE TREATED
BY A MOUSE GRANULOMA PROTEIN
OFF domain
Ford D;Hakansson H;
2006
773
IMP - some things achieved: much more to do
OFF domain
Forget J;Pavillon JF;Beliaeff B;Bocquene G;
1999
967
Joint action of pollutant combinations (pesticides and
metals) on survival (LC50 values) and acetylcholinesterase
activity of Tigriopus brevicornis (Copepoda, Harpacticoida)
ON experimental
study
Fortune JE;Armstrong DT;
1977
53
ANDROGEN PRODUCTION BY THECA AND GRANULOSA
ISOLATED FROM PROESTROUS RAT FOLLICLES
OFF domain
Fox JW;Olsen E;
2000
1006
Food web structure and the strength of transient indirect
effects
OFF domain
Fraysse B;Garric J;
2005
396
Prediction and experimental validation of acute toxicity of
beta-blockers in Ceriodaphnia dubia
OFF domain
Freed DH;
1997
529
Creating a table of authorization to empower staff
OFF domain
APPENDIX A
Authors
Page 16 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Frere C;Morange PE;Saut N;Tregouet
DA;Grosley M;Beltran J;Juhan-Vague I;Alessi
MC;
2005
800
Quantification of thrombin activatable fibrinolysis inhibitor
(TAR) gene polymorphism effects on plasma levels of TAR
measured with assays insensitive to isoform-dependent
artefact
OFF domain
Fricker SP;Slade E;Powell NA;
1999
709
Macrophage-derived superoxide is not required for nitric
oxide mediated tumour cell killing by RAW 264 cells
OFF domain
Frieling T;Wood JD;Cooke HJ;
1992
696
SUBMUCOSAL REFLEXES - DISTENSION-EVOKED IONTRANSPORT IN THE GUINEA-PIG DISTAL COLON
OFF domain
Frisbie DD;Sandler EA;Trotter GW;McIlwraith
CW;
2000
712
Metabolic and mitogenic activities of insulin-like growth
factor-1 in interleukin-1-conditioned equine cartilage
OFF domain
Fu P;Thompson JA;Bach LA;
2007
375
Promotion of cancer cell migration - An insulin-like growth
factor (IGF)-independent action of IGF-binding protein-6
OFF domain
Fuenzalida K;Quintanilla R;Ramos P;Piderit
D;Fuentealba RA;Martinez G;Inestrosa
NC;Bronfman M;
2007
748
Peroxisome proliferator-activated receptor gamma upregulates the Bcl-2 anti-apoptotic protein in neurons and
induces mitochondrial stabilization and protection against
oxidative stress and apoptosis
OFF domain
Fujita A;Fujieda S;Fukamichi K;
2006
593
Influence of partial substitution of Ce on the Curie
temperature and magnetic entropy change in itinerantelectron metamagnetic La(FexSi1-x)(13) compounds
OFF domain
Fujiwara K;Maekawa F;Dezaki K;Nakata
M;Yashiro T;Yada T;
2007
897
Oleic acid glucose-independently stimulates glucagon
secretion by increasing cytoplasmic Ca2+ via endoplasmic
reticulum Ca2+ release and Ca2+ influx in the rat islet
alpha-cells
OFF domain
Funfak A;Cao J;Knauer A;Martin K;Koehler JM;
2011
373
Synergistic effects of metal nanoparticles and a phenolic
uncoupler using microdroplet-based two-dimensional
approach
ON experimental
study
Fung SC;Fillenz M;
1984
166
MULTIPLE EFFECTS OF DRUGS ACTING ON
BENZODIAZEPINE RECEPTORS
OFF domain
Fuxa JR;
1979
78
INTERACTIONS OF THE MICROSPORIDIUM
VAIRIMORPHA-NECATRIX WITH A BACTERIUM, VIRUS,
AND FUNGUS IN HELIOTHIS-ZEA
OFF domain
G. L. Drusano;D. Z. D'Argenio;W. Symonds;P.
A. Bilello;J. McDowell;B. Sadler;A. Bye;J. A.
Bilello
1998
1137
Nucleoside analog 1592U89 and human immunodeficiency
virus protease inhibitor 141W94 are synergistic in vitro
ON experimental
study
Galazyn-Sidorczuk M;Brzoska MM;Jurczuk
M;Moniuszko-Jakoniuk J;
2009
1022
Oxidative damage to proteins and DNA in rats exposed to
cadmium and/or ethanol
OFF domain
Galhaup C;Haltrich D;
2001
1032
Enhanced formation of laccase activity by the white-rot
fungus Trametes pubescens in the presence of copper
OFF domain
Gallagher AP;Burnett AK;Bowen DT;Darley RL;
1998
785
Mutant RAS selectively promotes sensitivity of myeloid
leukemia cells to apoptosis by a protein kinase Cdependent process
OFF domain
Gallin EK;
1981
109
CALCIUM SPIKES IN CULTURED HUMAN RETICULAR
CELLS FROM PERITONEAL EXUDATES
OFF domain
Gao KM;Li P;
1993
702
POSTSYNAPTIC ACTIVITY EVOKED IN THE ROSTRAL
VENTROLATERAL MEDULLARY NEURONS BY
STIMULATION OF THE DEFENSE AREAS OF
HYPOTHALAMUS AND MIDBRAIN IN THE RAT
OFF domain
Garavini C;Cristofori M;
1984
167
THE EFFECT OF 5-ALPHA-DIHYDROTESTOSTERONE AND
5-BETA-DIHYDROTESTOSTERONE ON ERYTHROPOIESIS
OF THE NEWT, TRITURUS-CRISTATUS-CARNIFEX (LAUR)
OFF domain
Garcia J;Gamboaaldeco R;Stefani E;
1990
303
CHARGE MOVEMENT AND CALCIUM CURRENTS IN
SKELETAL-MUSCLE FIBERS ARE ENHANCED BY GTPGAMMA-S
OFF domain
Gasser J;Paganetti P;Carafoli E;Chiesi M;
1988
259
HETEROGENEOUS DISTRIBUTION OF CALMODULINOFF domain
DEPENDENT AND CAMP-DEPENDENT REGULATION OF CA2+ UPTAKE IN CARDIAC SARCOPLASMIC-RETICULUM
SUBFRACTIONS
APPENDIX A
Authors
Page 17 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Gay JL;Martin M;Ball E;
1985
187
THE IMPERMEABILITY OF POWDERY MILDEW CONIDIA
AND THEIR GERMINATION IN ARID ENVIRONMENTS
OFF domain
Gehan EA;
1982
136
PROGRESS OF THERAPY IN ACUTE-LEUKEMIA 19481981 - RANDOMIZED VERSUS NON-RANDOMIZED
CLINICAL-TRIALS
OFF domain
Gehring T;Krapohl S;
2007
921
Supranational regulatory agencies between independence
and control: the EMEA and the authorization of
pharmaceuticals in the European Single Market
OFF domain
Gennari C;Agnusdei D;Nardi P;Civitelli R;
1990
304
ESTROGEN PRESERVES A NORMAL INTESTINAL
RESPONSIVENESS TO 1,25-DIHYDROXYVITAMIN-D3 IN
OOPHORECTOMIZED WOMEN
OFF domain
Genolet R;Kersten S;Braissant O;Mandard
S;Tan NS;Bucher P;Desvergne B;Michalik
L;Wahli W;
2005
594
Promoter rearrangements cause species-specific hepatic
regulation of the glyoxylate reductase/hydroxypyruvate
reductase gene by the peroxisome proliferator-activated
receptor
OFF domain
Germolec DR;Kashon M;Nyska A;Kuper
CF;Portier C;Kommineni C;Johnson KA;Luster
MI;
2004
842
The accuracy of extended histopathology to detect
immunotoxic chemicals
OFF domain
Gessner PK;
1995
648
Isobolographic analysis of interactions: An update on
applications and utility
ON experimental
study
Ghelardini C;Galeotti N;Vettori AP;Capaccioli
S;Quattrone A;Bartolini A;
1997
969
Effect of K+ channel modulation on mouse feeding
behaviour
OFF domain
Giambalvo CT;Becker RE;
1981
110
MODULATORS OF MONOAMINE-OXIDASE IN PLASMA
OFF domain
Gianvittorio R;Restuccia A;Stephany J;
1995
614
A NEW ACTION PRINCIPLE FOR WITTENS TOPOLOGICAL
FIELD-THEORY
OFF domain
Gilman CP;Chan SL;Guo ZH;Zhu XX;Greig
N;Mattson MP;
2003
376
p53 is present in synapses where it mediates
mitochondrial dysfunction and synaptic degeneration in
response to DNA damage, and oxidative and excitotoxic
insults
OFF domain
Goel V;Pietrasik J;Matyjaszewski
K;Krishnamoorti R;
2010
973
Linear Viscoelasticity of Spherical SiO2 NanoparticleTethered Poly(butyl acrylate) Hybrids
OFF domain
Gokin AP;Jennings LJ;Mawe GM;
1996
844
Actions of calcitonin gene-related peptide in guinea pig
gallbladder ganglia
OFF domain
Gold D;Nissimov L;Lengy J;Keisari Y;
1994
580
ANTI-SCHISTOSOMULAR ACTIVITY OF HUMAN
MONOCYTES/MACROPHAGES IN RESPONSE TO
INTERLEUKIN-3 AND GRANULOCYTE-MACROPHAGE
COLONY-STIMULATING FACTOR STIMULATION
OFF domain
Golomb E;Hill MR;Brown RG;Keiser HR;
1994
794
OUABAIN ENHANCES THE MITOGENIC EFFECT OF SERUM OFF domain
IN VASCULAR SMOOTH-MUSCLE CELLS
Gomez-Eyles JL;Svendsen C;Lister L;Martin
H;Hodson ME;Spurgeon DJ;
2009
1125
Measuring and modelling mixture toxicity of imidacloprid
and thiacloprid on Caenorhabditis elegans and Eisenia
fetida
ON experimental
study
Gomezpuy.A;Sandoval F;Tuena M;Chavez
E;Pena A;
1969
11
K+ INDEPENDENT ACTION OF VALINOMYCIN IN
MITOCHONDRIA
OFF domain
Gordge MP;Hothersall JS;Noronha-Dutra AA;
1998
356
Evidence for a cyclic GMP-independent mechanism in the
anti-platelet action of S-nitrosoglutathione
OFF domain
Gougerotpocidalo MA;Fay M;Pocidalo JJ;
1984
168
INVIVO NORMOBARIC OXYGEN EXPOSURE DEPRESSES
OFF domain
SPLEEN-CELL INVITRO CON-A RESPONSE - EFFECTS OF 2MERCAPTOETHANOL AND PERITONEAL-CELLS
Grace JK;
1990
305
EFFECT OF ANTIOXIDANTS ON EASTERN
OFF domain
SUBTERRANEAN TERMITE (ISOPTERA,
RHINOTERMITIDAE) ORIENTATION TO FUNGAL EXTRACT
Graf M;Amrein TM;Graf S;Szalay R;Escher
F;Amado R;
2006
867
Reducing the acrylamide content of a semi-finished biscuit
on industrial scale
OFF domain
Gray LE;Rider CV;Howdeshell KL;Hotchkiss
AK;Wilson VS;Foster PMD;Furr J;
2009
345
Cumulative Effects of Administration Mixtures of
'Antiandrogens' in Rats: A New Framework Based upon
Common Systems Rather Than Common Mechanisms
ON trivial
APPENDIX A
Authors
Page 18 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Greene E;Levinson D;
1994
988
ANGULAR INDUCTION AS A FUNCTION OF THE LENGTH
AND POSITION OF SEGMENTS AND GAPS
OFF domain
Greenway CV;Murthy VS;
1971
14
MESENTERIC VASOCONSTRICTION AFTER ENDOTOXIN
ADMINISTRATION IN CATS PRETREATED WITH ASPIRIN
OFF domain
Gregoraszczuk EL;Rak A;Kawalec K;Ropstad
E;
2008
971
Steroid secretion following exposure of ovarian follicular
cells to single congeners and defined mixture of
polybrominateddibenzoethers (PBDEs), p,p '-DDT and its
metabolite p,p '-DDE
ON experimental
study
Griffin DE;Johnson RT;Tamashiro VG;Moench
TR;Jauregui E;Lindodesoriano I;Vaisberg A;
1987
233
INVITRO STUDIES OF THE ROLE OF MONOCYTES IN THE
IMMUNOSUPPRESSION ASSOCIATED WITH NATURAL
MEASLES-VIRUS INFECTIONS
OFF domain
Grimme LH;Faust M;Boedeker W;Altenburger
R;
1996
330
Aquatic toxicity of chemical substances in combination:
Still a matter of controversy
ON review
Grinstein S;Rothstein A;
1978
67
CHEMICALLY-INDUCED CATION PERMEABILITY IN REDCELL MEMBRANE-VESICLES - SIDEDNESS OF RESPONSE
AND PROTEINS INVOLVED
OFF domain
Gritlilinde A;Ruch JV;Linde A;
1995
757
POLYAMINE DEPLETION-MEDIATED EFFECTS ON
MURINE ODONTOGENESIS ARE DEPENDENT ON TOOTH
DEVELOPMENTAL STAGE AND CULTURE CONDITIONS
OFF domain
Grote M;Brack W;Walter HA;Altenburger R;
2005
920
Confirmation of cause-effect relationships using effectdirected analysis for complex environmental samples
ON experimental
study
Gruber R;Karreth F;Frommlet F;Fischer
MB;Watzek G;
2003
890
Platelets are mitogenic for periosteum-derived cells
OFF domain
Guo JF;Cong L;Rybin VO;Gertsberg
Z;Steinberg SF;
2010
571
Protein kinase C-delta regulates the subcellular localization
of Shc in H2O2-treated cardiomyocytes
OFF domain
Gupta GS;Bawa SR;
1978
68
RADIATION EFFECTS ON TESTES .13. STUDIES ON
ISOCITRATE DEHYDROGENASES FOLLOWING PARTIALBODY GAMMA-IRRADIATION
OFF domain
Gupta RS;Singh B;Stetsko DK;
1985
188
INHIBITION OF METABOLIC COOPERATION BY PHORBOL
ESTERS IN A CELL-CULTURE SYSTEM BASED ON
ADENOSINE KINASE DEFICIENT MUTANTS OF V79 CELLS
OFF domain
H. Yan;B. Zhang;S. Li;Q. C. A. Zhao
2010
1157
A formal model for analyzing drug combination effects and
its application in TNF-alpha-induced NF kappa B pathway
ON method dev.
Haag JD;Shepel LA;Kolman BA;Monson
DM;Benton ME;Watts KT;Waller JL;LopezGuajardo CC;Samuelson DJ;Gould MN;
2003
653
Congenic rats reveal three independent Copenhagen
alleles within the Mcs1 quantitative trait locus that confer
resistance to mammary cancer
OFF domain
Haas CN;Kersten SP;Wright K;Frank
MJ;Cidambi K;
1997
636
Generalization of independent response model for toxic
mixtures
ON method dev.
Hagiwara N;Sekijima Y;Hattori T;Hashimoto
T;Ikeda Si;
2003
1097
Two patients with different types of vasculitic neuropathy-- OFF domain
a comparison between cutaneous polyarteritis nodosa and
nonsystemic vasculitic neuropathy
Haider M;Kundi M;Grollknapp E;Koller M;
1990
306
INTERACTIONS BETWEEN NOISE AND AIR-POLLUTION
OFF domain
Halevy J;Boulpaep EL;Budinger ME;Binder
HJ;Hayslett JP;
1988
260
GLUCOCORTICOIDS HAVE A DIFFERENT ACTION THAN
ALDOSTERONE ON TARGET TISSUE
OFF domain
Hallett MB;Luzio JP;Campbell AK;
1981
111
STIMULATION OF CA2+-DEPENDENT CHEMILUMINESCENCE IN RAT POLYMORPHONUCLEAR
LEUKOCYTES BY POLYSTYRENE BEADS AND THE
NONLYTIC ACTION OF COMPLEMENT
OFF domain
Hamm RJ;
1977
54
TEST FOR RESPONSE SUMMATION WITH KEYPROJECTED STIMULI
OFF domain
Hamm RJ;Meltzer D;
1973
26
TEST FOR BOTH POSITIVE AND NEGATIVE RESPONSE
SUMMATION IN PIGEON
OFF domain
Han DH;Cha SY;Yang HY;
2004
366
Improvement of oxidative decomposition of aqueous
phenol by microwave irradiation in UV/H2O2 process and
kinetic study
OFF domain
Handler JA;Thurman RG;
1985
189
FATTY ACID-DEPENDENT ETHANOL-METABOLISM
OFF domain
APPENDIX A
Authors
Page 19 of 66
PubYear
ID
Title
CFT/EFSA/PPR/2010/02
Relevance
Hardy DB;Janowski BA;Corey DR;Mendelson
CR;
2006
820
Progesterone receptor plays a major antiinflammatory role OFF domain
in human myometrial cells by antagonism of nuclear factorkappa B activation of cyclooxygenase 2 expression
Harper MJK;Jones MA;Norris CJ;Woodard DS;
1989
284
PROSTAGLANDIN SYNTHESIS BY DAY-6 RABBIT
BLASTOCYSTS INVITRO
Harris D;Chuderland D;Bonfil D;Kraus
S;Seger R;Naor Z;
2003
397
Extracellular signal-regulated kinase and c-Src, but not Jun OFF domain
N-terminal kinase, are involved in basal and gonadotropinreleasing hormone-stimulated activity of the glycoprotein
hormone alpha-subunit promoter
Hatcher VB;Lazarus GS;Levine N;Burk
PG;Yost FJ;
1977
55
CHARACTERIZATION OF A CHEMOTACTIC AND
CYTOTOXIC PROTEINASE FROM HUMAN-SKIN
OFF domain
Hayward CR;
1998
754
De-facing power
OFF domain
Hazlerigg DG;Gonzalezbrito A;Lawson
W;Hastings MH;Morgan PJ;
1993
600
PROLONGED EXPOSURE TO MELATONIN LEADS TO TIME- OFF domain
DEPENDENT SENSITIZATION OF ADENYLATE-CYCLASE
AND DOWN-REGULATES MELATONIN RECEPTORS IN
PARS TUBERALIS CELLS FROM OVINE PITUITARY
Heinemann-Gruder A;
2003
891
Armies and politics - On the democratic control of east
European armies
OFF domain
Heinrich D;Stephinger U;Muellereckhardt C;
1977
56
SPECIFIC INTERACTION OF HLA ANTIBODIES (ELUATES)
WITH WASHED PLATELETS
OFF domain
Helmich RC;Aarts E;de Lange FP;Bloem
BR;Toni I;
2009
658
Increased Dependence of Action Selection on Recent
Motor History in Parkinson's Disease
OFF domain
Henry TB;Black MC;
2007
810
Mixture and single-substance acute toxicity of selective
serotonin reuptake inhibitors in Ceriodaphnia dubia
ON experimental
study
Hernandez-Balada E;Taylor MM;Phillips
JG;Marmer WN;Brown EM;
2009
482
Properties of biopolymers produced by transglutaminase
treatment of whey protein isolate and gelatin
OFF domain
Hesler LS;Oraze MJ;Grigarick AA;Palrang AT;
2000
1096
Numbers of rice water weevil larvae (Coleoptera :
Curculionidae) and rice plant growth in relation to adult
infestation levels and broadleaf herbicide applications
OFF domain
Hickman CS;
1981
112
EVOLUTION AND FUNCTION OF ASYMMETRY IN THE
ARCHEOGASTROPOD RADULA
OFF domain
Higashi Y;Holder K;Delafontaine P;
2010
1039
Thiazolidinediones Up-regulate Insulin-like Growth Factor1 Receptor via a Peroxisome Proliferator-activated
Receptor gamma-Independent Pathway
OFF domain
Higson FK;Jones OTG;
1984
169
OXYGEN RADICAL PRODUCTION BY HORSE AND PIG
NEUTROPHILS INDUCED BY A RANGE OF CRYSTALS
OFF domain
Hilf R;Feldstein ML;Gibson SL;Savlov ED;
1982
137
A LOGISTIC MODEL BASED ON ENZYME-ACTIVITIES FOR
THE PREDICTION OF RESPONSE OF BREAST-CANCER
PATIENTS TO CHEMOTHERAPY
OFF domain
Hirschelscholz S;Caverzasio J;Bonjour JP;
1985
190
INHIBITION OF PARATHYROID-HORMONE SECRETION
AND PARATHYROID HORMONE-INDEPENDENT
DIMINUTION OF TUBULAR CALCIUM REABSORPTION BY
WR-2721, A UNIQUE HYPOCALCEMIC AGENT
OFF domain
Hobfoll SE;Schroder KEE;Wells M;Malek M;
2002
1098
Communal versus individualistic construction of sense of
mastery in facing life challenges
OFF domain
Hochman S;Gutman Y;
1974
29
LITHIUM - ADH ANTAGONISM AND ADH INDEPENDENT
ACTION IN RATS WITH DIABETES-INSIPIDUS
OFF domain
Hodges G;Roberts DW;Marshall SJ;Dearden
JC;
2006
768
Defining the toxic mode of action of esther sulphonates
using the joint toxicity of mixtures
ON experimental
study
Hoffman WP;Leurgans SE;
1990
307
LARGE SAMPLE PROPERTIES OF 2 TESTS FOR
INDEPENDENT JOINT ACTION OF 2 DRUGS
ON trivial
Hoffmann A;Zureck A;Strey A;
1986
641
STIMULATION OF BRUCELLA CLEARANCE IN RATS AFTER
TREATMENT WITH BCG
OFF domain
Hogan RD;Franklin TD;Avery KS;Burke KM;
1982
138
ARTERIOLAR VASOCONSTRICTION IN RAT CREMASTER
MUSCLE INDUCED BY LOCAL HEAT-STRESS
OFF domain
OFF domain
APPENDIX A
Authors
Page 20 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Hohage H;Hess K;Jahl C;Greven J;Schlatter E;
1997
795
Renal and blood pressure effects of moxonidine and
clonidine in spontaneously hypertensive rats
OFF domain
Hohm O;Hull C;Zwiebach B;
2010
520
Background independent action for double field theory
OFF domain
Hoke F;
1995
506
BIOTECH COMPANIES SET TO PROFIT FROM ANIMALORGAN TRANSPLANTS
OFF domain
Hollshwandner CH;
1983
154
MODELS IN ADMINISTRATION AND THE DEANS
EXERCISE OF POWER AND LEADERSHIP
OFF domain
Holmes AM;Roderick HL;McDonald
F;Bootman MD;
2007
546
Interaction between store-operated and arachidonateactivated calcium entry
OFF domain
Holtug K;Laverty G;Arnason SS;Skadhauge E;
2009
542
NH4+ secretion in the avian colon. An actively regulated
barrier to ammonium permeation of the colon mucosa
OFF domain
Hopper R;Lancaster B;Garthwaite J;
2004
708
On the regulation of NMDA receptors by nitric oxide
OFF domain
Hoppner W;Sussmuth W;Obrien C;Seitz HJ;
1986
213
COOPERATIVE EFFECT OF THYROID AND
GLUCOCORTICOID HORMONES ON THE INDUCTION OF
HEPATIC PHOSPHOENOLPYRUVATE CARBOXYKINASE
INVIVO AND IN CULTURED-HEPATOCYTES
OFF domain
Hoque KM;Pal A;Nair GB;Chattopadhyay
S;Chakrabarti MK;
2001
602
Evidence of calcium influx across the plasma membrane
depends upon the initial rise of cytosolic calcium with
activation of IP3 in rat enterocytes by heat-stable
enterotoxin of Vibrio cholerae non-O1
OFF domain
Horst Grimme L;Altenburger R;Backhaus
T;Faust M;Boedeker W;Scholze M;
2000
523
Combined effects of environmental pollutants in
ecotoxicology: Biometrical models as concepts for
prediction and their experimental proof
ON method dev.
Hosley MA;Hughes SE;Oakley B;
1987
234
NEURAL INDUCTION OF TASTE-BUDS
OFF domain
Hotchkiss AK;Rider CV;Furr J;Howdeshell
KL;Blystone CR;Wilson VS;Gray LE;
2010
808
In utero exposure to an AR antagonist plus an inhibitor of
fetal testosterone synthesis induces cumulative effects on
F1 male rats
ON experimental
study
Howard GJ;Webster TF;
2007
342
Contrasting theories of additivity, synergy, and
antagonism in epidemiology and toxicology
ON trivial
Hsieh HL;Schafer BW;Weigle B;Heizmann CW;
2004
923
S100 protein translocation in response to extracellular
S100 is mediated by receptor for advanced glycation
endproducts in human endothelial cells
OFF domain
Hsu FL;Chen YC;Cheng JT;
2000
860
Caffeic acid as active principle from the fruit of Xanthium
strumarium to lower plasma glucose in diabetic rats
OFF domain
Hsu FL;Lai CW;Cheng JT;
1997
745
Antihyperglycemic effects of paeoniflorin and 8debenzoylpaeoniflorin, glucosides from the root of Paeonia
lactiflora
OFF domain
Hu JS;Vinson SB;
1997
526
In vitro development of Campoletis sonorensis (Hym.:
Ichneumonidae), a larval endoparasitoid of Heliothis
virescens (Lep.: Noctuidae) in an artificial medium with
insect sources from egg to third larval instar
OFF domain
Hu RR;Lager E;guilar-Aguilar A;Liu JZ;Lam
JWY;Sung HHY;Williams ID;Zhong YC;Wong
KS;Pena-Cabrera E;Tang BZ;
2009
473
Twisted Intramolecular Charge Transfer and AggregationInduced Emission of BODIPY Derivatives
OFF domain
Huber DM;Bendixen AC;Pathrose P;Srivastava
S;Dienger KM;Shevde NK;Pike JW;
2001
428
Androgens suppress osteoclast formation induced by
RANKL and macrophage-colony stimulating factor
OFF domain
Huff MW;Miller DB;Wolfe BM;Connelly
PW;Sawyez CG;
1997
585
Uptake of hypertriglyceridemic very low density
lipoproteins and their remnants by HepG2 cells: the role of
lipoprotein lipase, hepatic triglyceride lipase, and cell
surface proteoglycans
OFF domain
Hughes A;Vaney DI;
1982
139
THE ORGANIZATION OF BINOCULAR CORTEX IN THE
PRIMARY VISUAL AREA OF THE RABBIT
OFF domain
Hughes EH;Schlichtenbrede FC;Murphy
CC;Broderick C;Van Rooijen N;Ali RR;Dick AD;
2004
774
Minocycline delays photoreceptor death in the rds mouse
through a microglia-independent mechanism
OFF domain
APPENDIX A
Authors
Page 21 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Hughes HPA;Campos M;Littelvandenhurk
SVD;Zamb T;Sordillo LM;Godson D;Babiuk LA;
1992
640
MULTIPLE ADMINISTRATION WITH INTERLEUKIN-2
POTENTIATES ANTIGEN-SPECIFIC RESPONSES TO
SUBUNIT VACCINATION WITH BOVINE HERPESVIRUS-1
GLYCOPROTEIN-IV
OFF domain
Hughes JT;
1998
944
The role of development agencies in regional policy: An
academic and practitioner approach
OFF domain
Huigen YM;Krips HJ;Hulleman S;Gelsema
WJ;De Ligny CL;
1990
948
The adsorption of 99mTc(Sn)-diphosphonate complexes
on tri-calciumphosphate: the influence of preparation
conditions, ligand-type, incubation media and adsorption
conditions. The reversibility of the adsorption
OFF domain
Hunyor SN;Bauer GE;Ross M;Larkin H;
1980
91
LABETALOL AND PROPRANOLOL IN MILD
HYPERTENSIVES - COMPARISON OF BLOOD-PRESSURE
AND PLASMA-VOLUME EFFECTS
OFF domain
Hutmacher MM;Chapel S;Agin MA;Fleishaker
JC;Lalonde RL;
2008
957
Performance characteristics for some typical QT study
designs under the ICH E-14 guidance
OFF domain
Huynh HT;Tetenes E;Wallace L;Pollak M;
1993
834
INVIVO INHIBITION OF INSULIN-LIKE GROWTH FACTORI GENE-EXPRESSION BY TAMOXIFEN
OFF domain
Hwang JM;Yamanouchi J;Santamaria P;Kubes
P;
2004
503
A critical temporal window for selectin-dependent CD4(+)
lymphocyte homing and initiation of late-phase
inflammation in contact sensitivity
OFF domain
Iatsenko NM;Tsintsadze TS;Lozova NO;
2007
692
The synthetic cannabinoid analog WIN 55,212-2
potentiates the amplitudes of glycine-activated currents
OFF domain
Ide BA;Tobias C;Kay M;Monk J;De Zapien JG;
1990
1045
A COMPARISON OF COPING STRATEGIES USED
EFFECTIVELY BY OLDER ANGLO AND MEXICANAMERICAN WIDOWS A LONGITUDINAL STUDY
OFF domain
Ikebuchi M;Kimura H;Hill CK;Aoyama T;
1995
1126
ARE 3 FORMS OF POTENTIALLY LETHAL DAMAGE
EXPRESSED AFTER X-IRRADIATION BY TREATMENT
WITH HYPERTONIC SOLUTIONS IN CHINESE-HAMSTER
V79 CELLS
OFF domain
Ikeda K;Sunose H;Takasaka T;
1996
1081
Effects of protein kinase C on the Na+-H+ exchange in
the cochlear outer hair cell
OFF domain
Illman RJ;Topping DL;Dowling K;Trimble
RP;Russell GR;Storer GB;
1991
589
EFFECTS OF SOLVENT-EXTRACTION ON THE
HYPOCHOLESTEROLEMIC ACTION OF OAT BRAN IN THE
RAT
OFF domain
Imaz MS;Schmelling MF;Kaempfer S;Spallek
R;Singh M;
2008
887
Serodiagnosis of tuberculosis: Specific detection of free
and complex-dissociated antibodies anti-Mycobacterium
tuberculosis recombinant antigens
OFF domain
Inge TH;Hoover SK;Susskind BM;Barrett
SK;Bear HD;
1992
1071
INHIBITION OF TUMOR-SPECIFIC CYTOTOXIC
LYMPHOCYTE-T RESPONSES BY TRANSFORMING
GROWTH FACTOR-BETA-1
OFF domain
Ingermann AR;Yang YF;Han JF;Mikami
A;Garza AE;Mohanraj L;Fan LB;Idowu
M;Ware JL;Kim HS;Lee DY;Oh Y;
2010
633
Identification of a Novel Cell Death Receptor Mediating
IGFBP-3-induced Anti-tumor Effects in Breast and Prostate
Cancer
OFF domain
Isaacs MA;Hatzios KK;Wilson HP;Toler J;
2006
569
Halosulfuron and 2,4-d mixtures' effects on common
Lambsquarters (Chenopodium album)
ON experimental
study
Ishizuka M;Yamada F;Tanaka Y;Takeuchi
Y;Imaseki H;
1991
625
SEQUENTIAL INDUCTION OF MESSENGER-RNAS FOR
PHENYLALANINE AMMONIA-LYASE IN SLICES OF
POTATO-TUBER
OFF domain
Ito H;Tsuchimochi H;Tada Y;Kurachi Y;
1997
422
Phosphorylation-independent inhibition by intracellular
cyclic nucleotides of brain inwardly rectifying K+ current
expressed in Xenopus oocytes
OFF domain
Iuliano A;Lecci C;Salvadori P;
2003
424
The s-triazine moiety as a scaffold for connecting different
chiral auxiliaries: synthesis of new biselector CSPs for
enantioselective chromatography
OFF domain
Ivanovski O;Szumilak D;Nguyen-Khoa
T;Nikolov IG;Joki N;Mothu N;Maizel
J;Westenfeld R;Ketteler M;Lacour B;Druke
TB;Massy ZA;
2008
779
Effect of simvastatin in apolipoprotein E deficient mice
with surgically induced chronic renal failure
OFF domain
APPENDIX A
Authors
Page 22 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Iwao M;
1972
22
Nursing technic based on independent action
OFF domain
Izumi H;Yallampalli C;Garfield RE;
1993
727
GESTATIONAL CHANGES IN L-ARGININE INDUCED
RELAXATION OF PREGNANT RAT AND HUMAN
MYOMETRIAL SMOOTH-MUSCLE
OFF domain
Izutsu KI;Yoshioka S;Kojima S;
1994
1005
PHYSICAL STABILITY AND PROTEIN STABILITY OF
FREEZE-DRIED CAKES DURING STORAGE AT ELEVATEDTEMPERATURES
OFF domain
J. Afeltra;R. G. Vitale;J. W. Mouton;P. E.
Verweij
2004
1129
Potent synergistic in vitro interaction between
nonantimicrobial membrane-active compounds and
itraconazole against clinical isolates of Aspergillus
fumigatus resistant to Itraconazole
ON trivial
J. Meletiadis;Dtat Dorsthorst;P. E. Verweij
2006
1146
The concentration-dependent nature of in vitro
ON experimental
amphotericin B-itraconazole interaction against Aspergillus study
fumigates: isobolographic and response surface analysis of
complex pharmacodynamic interactions
J. Meletiadis;J. W. Mouton;Jfgm Meis;P. E.
Verweij
2003
1148
In vitro drug interaction modeling of combinations of
azoles with terbinafine against clinical Scedospotium
prolificans isolates
ON experimental
study
J. Meletiadis;Jfgm Meis;J. W. Mouton;P. E.
Verweij
2002
1147
Methodological issues related to antifungal drug
interaction modelling for filamentous fungi
ON trivial
J. Meletiadis;P. E. Verweij;Dtat
Dorsthorst;Jfgm Meis;J. W. Mouton
2005
1150
Assessing in vitro combinations of antifungal drugs against ON experimental
yeasts and filamentous fungi: comparison of different drug study
interaction models
J. Meletiadis;V. Petraitis;R. Petraitiene;P.
Lin;T. Stergiopoulou;A. M. Kelaher;T. Sein;R.
L. Schaufele;J. Bacher;T. J. Walsh
2006
1149
Triazole-polyene antagonism in experimental invasive
pulmonary aspergillosis: In vitro and in vivo correlation
ON experimental
study
Jaffe CA;Turgeon DK;Lown K;mott-Friberg
R;Watkins PB;
2002
940
Growth hormone secretion pattern is an independent
regulator of growth hormone actions in humans
OFF domain
Jansen A;Hagen RM;Schmidt W;Ignatius
R;Perez-Canto A;Zeitz M;Schneider T;
2005
1116
Pulmonary melioidosis associated with bronchiolitis
obliterans organizing pneumonia
OFF domain
Jantas D;Szymanska M;Budziszewska
B;Lason W;
2009
676
An involvement of BDNF and PI3-K/Akt in the antiapoptotic effect of memantine on staurosporine-evoked
cell death in primary cortical neurons
OFF domain
Japelj I;Budihna MV;Stanovnik L;Krisch I;
1999
1000
Stereoselective and endothelium-independent action of
nicardipine on the isolated porcine coronary artery
OFF domain
Jaunet T;Notteghem JL;Rapilly F;
1996
815
Pathogenicity process of Pseudomonas fuscovaginae, the
causal agent of sheath brown rot of rice
OFF domain
Jehn UW;Karlin L;
1971
15
INDEPENDENT ACTION OF THYMUS AND BONE MARROW
CELLS DURING SECONDARY RESPONSE OF DIRECT
PLAQUE-FORMING CELLS
OFF domain
Jenne JW;Yeoh HC;Shaughnessy TK;Hur K;
1992
323
THEOPHYLLINE PRODUCES OVER-ADDITIVE
RELAXATION OF CANINE TRACHEAL SMOOTH-MUSCLE
WHEN COMBINED WITH BETA-AGONISTS - THE DOSERESPONSE RELATIONSHIP
ON trivial
Jennings JB;Gelder SR;
1976
47
OBSERVATIONS ON FEEDING MECHANISM, DIET AND
DIGESTIVE PHYSIOLOGY OF HISTRIOBDELLA-HOMARI
VANBENEDEN 1858 - ABERRANT POLYCHAETE
SYMBIOTIC WITH NORTH-AMERICAN AND EUROPEAN
LOBSTERS
OFF domain
Jensen BL;Ellekvist P;Skott O;
1999
882
Renin secretion from permeabilized juxtaglomerular cells
requires a permeant cation
OFF domain
Jensen D;Bayley M;Holmstrup M;
2009
827
Synergistic interaction between 4-nonylphenol and high
but not low temperatures in Dendrobaena octaedra
ON experimental
study
Jensen MC;Bunn HF;Halikas GC;Nathan DG;
1972
23
Oxygen affinity independent action of cyanate and 2,3
DPG on sickling
OFF domain
Jensen MS;Azouz R;Yaari Y;
1994
363
VARIANT FIRING PATTERNS IN RAT HIPPOCAMPAL
PYRAMIDAL CELLS MODULATED BY EXTRACELLULAR
POTASSIUM
OFF domain
APPENDIX A
Authors
Page 23 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Jensen MS;Azouz R;Yaari Y;
1996
359
Spike after-depolarization and burst generation in adult rat
hippocampal CA1 pyramidal cells
OFF domain
Ji RR;Yao HX;Sun XS;
2011
578
Actor-independent action search using spatiotemporal
vocabulary with appearance hashing
OFF domain
Johnson BR;Voigt R;Merrill CL;Atema J;
1991
1095
ACROSS-FIBER PATTERNS MAY CONTAIN A SENSORY
CODE FOR STIMULUS-INTENSITY
OFF domain
Johnson SH;Sprehn G;Saykin AJ;
2002
551
Intact motor imagery in chronic upper limb hemiplegics:
Evidence for activity-independent action representations
OFF domain
Jones BEH;Haynes RJ;Phillips IR;
2010
905
Effect of amendment of bauxite processing sand with
organic materials on its chemical, physical and microbial
properties
OFF domain
Jones GE;
2002
1023
Policy and power: one hundred years of local education
authorities in wales
OFF domain
Jonker MJ;Svendsen C;Bedaux JJM;Bongers
M;Kammenga JE;
2005
591
Significance testing of synergistic/antagonistic, dose leveldependent, or dose ratio-dependent effects in mixture
dose-response analysis
ON experimental
study
Jorge AAL;Souza S;Arnhold IJP;Mendonca BB;
2004
715
The first homozygous mutation (S226I) in the highlyconserved WSXWS-like motif of the GH receptor causing
Laron syndrome: supression of GH secretion by GnRH
analogue therapy not restored by dihydrotestosterone
administration
OFF domain
Joseph K;Baby TG;
1991
737
EARLY DEVELOPMENTAL PROFILE OF ORNITHINE
DECARBOXYLASE IN THE FROG, MICROHYLA-ORNATA
AND ITS REGULATION BY POLYAMINES
OFF domain
Juergen Suehnel
1998
1153
Parallel dose-response curves in combination experiments
ON experimental
study
Junejo IN;Dexter E;Laptev I;Perez P;
2011
1046
View-Independent Action Recognition from Temporal SelfSimilarities
OFF domain
Jung DS;Bodyfelt FW;Daeschel MA;
1992
1002
INFLUENCE OF FAT AND EMULSIFIERS ON THE EFFICACY
OF NISIN IN INHIBITING LISTERIA-MONOCYTOGENES IN
FLUID MILK
OFF domain
Junghans M;Backhaus T;Faust M;Scholze
M;Grimme LH;
2003
338
Toxicity of sulfonylurea herbicides to the green alga
Scenedesmus vacuolatus: Predictability of combination
effects
ON experimental
study
Junghans M;Backhaus T;Faust M;Scholze
M;Grimme LH;
2003
493
Predictability of combined effects of eight chloroacetanilide ON experimental
herbicides on algal reproduction
study
Junghans M;Backhaus T;Faust M;Scholze
M;Grimme LH;
2006
592
Application and validation of approaches for the predictive
hazard assessment of realistic pesticide mixtures
ON experimental
study
Jurczuk M;Brzoska MM;Moniuszko-Jakoniuk J;
2007
1021
Hepatic and renal concentrations of vitamins E and C in
lead- and ethanol-exposed rats. An assessment of their
involvement in the mechanisms of peroxidative damage
OFF domain
Jurczuk M;Brzoska MM;Rogalska J;MoniuszkoJakoniuk J;
2003
401
Iron body status of rats chronically exposed to cadmium
and ethanol
ON trivial
Jurczuk M;Moniuszko-Jakoniuk J;Brzoska MM;
2006
961
Involvement of some low-molecular thiols in the
peroxidative mechanisms of lead and ethanol action on rat
liver and kidney
OFF domain
Jurczuk M;Moniuszko-Jakoniuk J;Brzoska
MM;Roszczenko A;
2005
613
Vitamins E and C concentrations in the liver and kidney of
rats exposed to cadmium and ethanol
OFF domain
Jurczuk M;Moniuszko-Jakoniuk J;Rogalska J;
2006
845
Glutathione-related enzyme activity in liver and kidney of
rats exposed to cadmium and ethanol
OFF domain
Jurczuk M;Moniuszko-Jakoniuk J;Rogalska J;
2006
849
Evaluation of oxidative stress in hepatic mitochondria of
rats exposed to cadmium and ethanol
OFF domain
Kable PF;Jeffery H;
1980
92
SELECTION FOR TOLERANCE IN ORGANISMS EXPOSED
TO SPRAYS OF BIOCIDE MIXTURES - THEORETICALMODEL
OFF domain
Kaesemeyer WH;Caldwell RB;Huang
JZ;Caldwell RW;
1999
731
Pravastatin sodium activates endothelial nitric oxide
synthase independent of its cholesterol-lowering actions
OFF domain
APPENDIX A
Authors
Page 24 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Kaiser RA;Wiley C;Martin S;Buxton ILO;
2000
333
Characterization of myometrial. smooth muscle potassium
channels: Search for the cGMP-independent action of NO
OFF domain
Kalbitz K;Meyer A;Yang R;Gerstberger P;
2007
517
Response of dissolved organic matter in the forest floor to
long-term manipulation of litter and throughfall inputs
OFF domain
Kalla R;Bohatschek M;Kloss CUA;Krol J;von
Maltzan X;Raivich G;
2003
797
Loss of microglial ramification in microglia-astrocyte
cocultures: Involvement of adenylate cyclase, calcium,
phosphatase, and G(i)-protein systems
OFF domain
Kalugina AV;Komarova LN;Petin VG;
2002
549
Mathematical description of synergistic interactions of
environment temperature and microwaves for animal
warming
OFF domain
Kamaya H;Ueda I;Eyring H;
1976
48
COMBINED EFFECTS OF DISSOCIABLE AND
UNDISSOCIABLE LOCAL-ANESTHETICS UPON ATPINDUCED FIREFLY BIOLUMINESCENCE
OFF domain
Kanner J;Harel S;Shagalovich J;Berman S;
1984
170
ANTIOXIDATIVE EFFECT OF NITRITE IN CURED MEATPRODUCTS - NITRIC-OXIDE IRON COMPLEXES OF LOWMOLECULAR WEIGHT
OFF domain
Kapasi K;Albert SE;Yie SM;Zavazava
N;Librach CL;
2000
579
HLA-G has a concentration-dependent effect on the
generation of an allo-CTL response
OFF domain
Kaplan SR;Goslow GE;
1989
285
NEUROMUSCULAR ORGANIZATION OF THE PECTORALIS
(PARS-THORACICUS) OF THE PIGEON (COLUMBALIVIA) - IMPLICATIONS FOR MOTOR CONTROL
OFF domain
Karanth S;Lyson K;McCann SM;
1993
826
ROLE OF NITRIC-OXIDE IN INTERLEUKIN-2-INDUCED
CORTICOTROPIN-RELEASING FACTOR RELEASE FROM
INCUBATED HYPOTHALAMI
OFF domain
Karas CA;Picker M;Poling A;
1986
214
EFFECTS OF PHENOBARBITAL IN COMBINATION WITH
PHENYTOIN OR VALPROIC ACID ON THE DELAYEDMATCHING-TO-SAMPLE PERFORMANCE OF PIGEONS
ON trivial
Karg O;Bonnet R;Magnussen H;Kohler
D;Geiseler J;Haidl P;Mader I;Schucher B;
2004
801
Respiratory therapist: introduction of a new profession
OFF domain
Kashiwagura T;Sakurai K;Takeguchi N;
1987
235
PHYSIOLOGICAL ASSAY OF LIPOSOME-MEDIATED
TRANSPORT OF A DRUG ACROSS XENOPUS INTESTINE CELL LIPOSOME INTERACTION
OFF domain
Kaul K;Bhattacharya S;
1988
261
THYROID-HORMONE STIMULATION OF THE PERCH
(ANABAS-TESTUDINEUS BLOCH) OVARIAN
MITOCHONDRIAL STEROIDOGENESIS
OFF domain
Kaverina NV;Chichkanov GG;
2003
987
Pharmacology of antiarrhythmic drugs: New aspects
OFF domain
Kaverina NV;Lyskovtsev VV;Popova EP;
2007
914
Comparative study of the electrophysiological properties of
class III medicinals (cardiocyclide, nibentan, sotalol) and
evaluation of their efficacy in atrial fibrillation produced by
auricular and vagal stimulation
OFF domain
Kavok NS;Krasilnikova OA;Babenko NA;
2001
461
Thyroxine signal transduction in liver cells involves
phospholipase C and phospholipase D activation. genomic
independent action of thyroid hormone
OFF domain
Kay JN;Link BA;Baier H;
2005
607
Staggered cell-intrinsic timing of ath5 expression underlies
the wave of ganglion cell neurogenesis in the zebrafish
retina
OFF domain
Keeler R;
1982
140
ATRIAL NATRIURETIC FACTOR HAS A DIRECT,
PROSTAGLANDIN-INDEPENDENT ACTION ON KIDNEYS
OFF domain
Keledjian K;Kyprianou N;
2003
780
Anoikis induction by quinazoline based alpha 1adrenoceptor antagonists in prostate cancer cells:
Antagonistic effect of bcl-2
OFF domain
Kennedy JG;Breslow MJ;Tobin JR;Traystman
RJ;
1991
1063
CHOLINERGIC REGULATION OF ADRENAL-MEDULLARY
BLOOD-FLOW
OFF domain
Kerrigan JR;Martha PM;Krieg RJ;Queen
TA;Monahan PE;Rogol AD;
1991
1067
AUGMENTED HYPOTHALAMIC PROOPIOMELANOCORTIN
GENE-EXPRESSION WITH PUBERTAL DEVELOPMENT IN
THE MALE-RAT - EVIDENCE FOR AN ANDROGEN
RECEPTOR-INDEPENDENT ACTION
OFF domain
APPENDIX A
Authors
Page 25 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Keuren JFW;Cauwenberghs S;Heeremans
J;de Kort W;Heemskerk JWM;Curvers J;
2006
453
Platelet ADP response deteriorates in synthetic storage
media
OFF domain
Khan MJ;Scaife JR;Hovell FD;
1999
1122
The effect of different sources of urease enzyme on the
nutritive value of wheat straw treated with urea as a
source of ammonia
OFF domain
Kiefer J;Schreiber A;Gutermuth F;Koch
S;Schmidt P;
1999
698
Mutation induction by different types of radiation at the
Hprt locus
OFF domain
Kienle C;Koehler H;Gerhardt A;
2009
597
Behavioural and developmental toxicity of chlorpyrifos and
nickel chloride to zebrafish (Danio rerio) embryos and
larvae
ON experimental
study
Kiesel L;Catt KJ;
1987
236
STIMULATION OF LUTEINIZING-HORMONE RELEASE AND OFF domain
CYCLIC-NUCLEOTIDE PRODUCTION BY ARACHIDONICACID IN CULTURED PITUITARY GONADOTROPHS
Kim HJ;Lee Y;Chang EJ;Kim HM;Hong SP;Lee
ZH;Ryu J;Kim HH;
2007
699
Suppression of osteoclastogenesis by N,N-dimethyl-Derythro-sphingosine: A sphingosine kinase inhibitionindependent action
OFF domain
Kim HS;Ingermann AR;Tsubaki J;Twigg
SM;Walker GE;Oh Y;
2004
631
Insulin-like growth factor-binding protein 3 induces
caspase-dependent apoptosis through a death receptormediated pathway in MCF-7 human breast cancer cells
OFF domain
Kim KT;Lee YG;Kim SD;
2006
878
Combined toxicity of copper and phenol derivatives to
Daphnia magna: Effect of complexation reaction
ON experimental
study
Kimoto Y;Kinoshita H;Nakahata K;Dojo
M;Iranami H;Hatano Y;
2003
978
Mexiletine differentially modulates vasorelaxation
OFF domain
mediated by adenosine triphosphate-sensitive K+ channels
in aortas from normotensive and hypertensive rats
Kirchner H;Holden HT;Herberman RB;
1975
37
SPLENIC SUPPRESSOR MACROPHAGES INDUCED IN MICE
BY INJECTION OF CORYNEBACTERIUM PARVUM
OFF domain
Kitahara K;Kusunoki N;Kakiuchi T;Suguro
T;Kawai S;
2009
847
Adiponectin stimulates IL-8 production by rheumatoid
synovial fibroblasts
OFF domain
Kitamura T;Murayama T;Nomura Y;
2000
1084
Enhancement of Ca2+-induced noradrenaline release by
vanadate in PC12 cells: possible involvement of tyrosine
phosphorylation
OFF domain
Kiyatkin EA;Rebec GV;
2000
622
Dopamine-independent action of cocaine on striatal and
accumbal neurons
OFF domain
Klaunig JE;Ruch RJ;
1987
237
ROLE OF CYCLIC-AMP IN THE INHIBITION OF MOUSE
HEPATOCYTE INTERCELLULAR COMMUNICATION BY
LIVER-TUMOR PROMOTERS
OFF domain
Klinger V;Drezel G;Grokhmann G;Shteinert
K;Vol'kmann P;
1978
598
Modification of drug oxidation in rat liver microsomal
fractions by mitochondria
OFF domain
Klingler JP;Edwards OR;Singh KB;
2007
577
Independent action and contrasting phenotypes of
resistance genes against spotted alfalfa aphid and
bluegreen aphid in Medicago truncatula
OFF domain
Knauert S;Escher B;Singer H;Hollender
J;Knauer K;
2008
683
Mixture toxicity of three photosystem II inhibitors
ON experimental
(atrazine, isoproturon, and diuron) toward photosynthesis study
of freshwater phytoplankton studied in outdoor mesocosms
Koch CC;Dasgupta A;Kroeger DM;Scarbrough
JO;
1979
79
PEAK EFFECT, SUMMATION PROBLEM, AND MAGNETIC
HISTORY IN A SUPERCONDUCTING NB-38 AT . PERCENT
HF ALLOY
OFF domain
Koehler E;Varnai G;Knospe S;Foerster
W;Michaelis D;Wanka H;
1991
900
EFFECTS OF THE PLATELET-ACTIVATING FACTOR
ANTAGONIST BN 52021 ON ANTI-ISLET CYTOTOXICITY
OF MONONUCLEAR CELLS AND SERUM FROM TYPE 1
INSULIN-DEPENDENT DIABETIC PATIENTS
OFF domain
Koenigsberger R;Parsons SM;
1980
93
BICARBONATE AND MAGNESIUM ION-ATP DEPENDENT
STIMULATION OF ACETYLCHOLINE UPTAKE BY TORPEDO
ELECTRIC ORGAN SYNAPTIC VESICLES
OFF domain
Koessler F;Nasledov GA;Shvinka N;
1996
525
The effects of calcium channel modulators on contractions
of tonic frog muscle fibres
OFF domain
APPENDIX A
Authors
Page 26 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Kofalvi A;Vizi ES;Ledent C;Sperlagh B;
2003
616
Cannabinoids inhibit the release of H-3 glutamate from
rodent hippocampal synaptosomes via a novel CB1
receptor-independent action
OFF domain
Kojo M;Ogawa T;Sonoda H;Yamada K;
1995
870
Development changes in carotid artery blood flow
waveform by component analysis in children
OFF domain
Komatsu M;Yajima H;Yamada S;Kaneko
T;Sato Y;Yamauchi K;Hashizume K;Aizawa T;
1999
567
Augmentation of Ca2+-stimulated insulin release by
glucose and long-chain fatty acids in rat pancreatic islets Free fatty acids mimic ATP-sensitive K+ channelindependent insulinotropic action of glucose
OFF domain
Kondaiah N;Panda B;
1989
286
EFFECT OF PHOSPHATE AND SPENT HEN YOLK ON THE
QUALITY OF CHICKEN SAUSAGES FROM SPENT HENS
OFF domain
Kongsuphol P;Hieke B;Ousingsawat J;Almaca
J;Viollet B;Schreiber R;Kunzelmann K;
2009
764
Regulation of Cl- secretion by AMPK in vivo
OFF domain
Koopman JS;Weed DL;
1990
308
EPIGENESIS THEORY - A MATHEMATICAL-MODEL
RELATING CAUSAL CONCEPTS OF PATHOGENESIS IN
INDIVIDUALS TO DISEASE PATTERNS IN POPULATIONS
OFF domain
Korn EL;
1985
191
MODELS OF INDEPENDENT ACTION FOR COMBINATION
THERAPY
ON trivial
Kornel L;Manisundaram B;Nelson WA;
1993
942
GLUCOCORTICOIDS REGULATE NA+ TRANSPORT IN
VASCULAR SMOOTH-MUSCLE THROUGH THE
GLUCOCORTICOID RECEPTOR-MEDIATED MECHANISM
OFF domain
Kortenkamp A;Faust M;Scholze M;Backhaus T;
2007
449
Low-Level Exposure to Multiple Chemicals: Reason for
Human Health Concerns?
ON review
Krantz J;Tuomas R;Bhushan B;
1999
528
Electrorheological fluids for lubrication (c)
OFF domain
Krieglstein J;Lippert K;Poch G;
1996
372
Apparent independent action of nimodipine and glutamate OFF domain
antagonists to protect cultured neurons against glutamateinduced damage
Krishnan K;Paterson J;Williams DT;
1997
985
Health risk assessment of drinking water contaminants in
Canada: The applicability of mixture risk assessment
methods
ON review
Kroger N;Frenzel H;
1993
325
RESISTANCE TO DIURETICS IN CONGESTIVE-HEARTFAILURE
OFF domain
Krohn MH;Hellmann JR;Shelleman DL;Green
DJ;Sakoske GE;Salem JA;
2002
514
Test methodology for strength testing of soda-lime-silica
float glass before and after enameling
OFF domain
Kubes P;Sihota E;Hickey MJ;
1997
627
Endogenous but not exogenous nitric oxide decreases TNF- OFF domain
alpha-induced leukocyte rolling
Kudeken N;Kawakami K;Saito A;
2000
1100
Mechanisms of the in vitro fungicidal effects of human
neutrophils against Penicillium marneffei induced by
granulocyte-macrophage colony-stimulating factor (GMCSF)
OFF domain
Kulkarni AP;
1976
49
JOINT ACTION OF INSECTICIDES AGAINST HOUSEFLIES
OFF domain
Kumagai Y;Ikeda Y;Toki S;
1992
364
HYDROXYL RADICAL-MEDIATED CONVERSION OF
MORPHINE TO MORPHINONE
OFF domain
Kungolos A;Emmanouil C;Tsiridis
V;Tsiropoulos N;
2009
1059
Evaluation of toxic and interactive toxic effects of three
agrochemicals and copper using a battery of microbiotests
ON experimental
study
Kunz PY;Fent K;
2006
1093
Estrogenic activity of UV filter mixtures
ON experimental
study
Kurashige N;Hiramatsu N;Ohkawa
K;Yakushijin T;Kiso S;Kanto T;Takehara
T;Kasahara A;Doi Y;Yamada A;Oshita M;Mita
E;Hagiwara H;Nagase T;Yoshihara H;Hayashi
E;Imai Y;Kato M;Kashihara T;Hayashi N;
2009
862
Factors contributing to antiviral effect of adefovir dipivoxil
OFF domain
therapy added to ongoing lamivudine treatment in patients
with lamivudine-resistant chronic hepatitis B
Ladics GS;Kawabata TT;White KL;
1991
321
SUPPRESSION OF THE INVITRO HUMORAL IMMUNERESPONSE OF MOUSE SPLENOCYTES BY 7,12DIMETHYLBENZ A ANTHRACENE METABOLITES AND
INHIBITION OF IMMUNOSUPPRESSION BY ALPHANAPHTHOFLAVONE
OFF domain
APPENDIX A
Authors
Page 27 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Lafond J;Bouvier C;Forget H;Lagace
G;Kaufman S;Collu R;
1989
287
REGULATION OF BASAL AND STIMULATED PROLACTINRELEASE IN PROLACTIN-SECRETING RAT PITUITARYTUMORS
OFF domain
Lafourcade CA;Zhang L;Alger BE;
2009
460
Novel mGluR- and CB1R-Independent Suppression of
GABA Release Caused by a Contaminant of the Group I
Metabotropic Glutamate Receptor Agonist, DHPG
OFF domain
Lakshminarayanan S;Antonetti DA;Gardner
TW;Tarbell JM;
2000
776
Effect of VEGF on retinal microvascular endothelial
hydraulic conductivity: The role of NO
OFF domain
Lam GKY;
1989
288
ANALYSIS OF INTERACTION FOR MIXTURES OF AGENTS
USING THE LINEAR ISOBOLE
ON trivial
Lam GKY;
1994
426
A GENERAL FORMULATION OF THE CONCEPT OF
INDEPENDENT ACTION FOR THE COMBINED EFFECTS OF
AGENTS
ON review
Lam GKY;
1995
1040
THE DEFINITION OF DAMAGE INTERACTION IN
INTERNATIONAL-COMMISSION-OF-RADIATION-UNITSAND-MEASUREMENT REPORT-30
OFF domain
Lambert JC;Lipscomb JC;
2007
655
Mode of action as a determining factor in additivity models
for chemical mixture risk assessment
ON review
Lambert JC;Zhou ZX;Kang YJ;
2002
336
Zinc inhibition of ethanol-induced liver apoptosis is
independent of metallothionein protection
OFF domain
Langeland N;Haarr L;Holmsen H;
1986
215
EVIDENCE THAT NEOMYCIN INHIBITS HSV-1 INFECTION
OF BHK CELLS
OFF domain
Langley RS;
1997
1058
The response of two-dimensional periodic structures to
impulsive point loading
OFF domain
Lankin VZ;Gordeeva NT;Tikhaze AK;Vikhert
AM;
1985
610
ANIMAL LIPOXYGENASES THE NATURE OF SUBSTRATE
AND CHANGES IN THE CONFORMATION OF
RETICULOCYTE LIPOXYGENASE DURING ITS
INTERACTION WITH MEMBRANES
OFF domain
Larkman PM;Kelly JS;
1997
884
Modulation of I-H by 5-HT in neonatal rat motoneurones
in vitro: Mediation through a phosphorylation independent
action of cAMP
OFF domain
Laserna EJ;Luz Valero M;Sanz L;Sanchez del
Pino MM;Calvete JJ;Barettino D;
2009
805
Proteomic Analysis of Phosphorylated Nuclear Proteins
Underscores Novel Roles for Rapid Actions of Retinoic Acid
in the Regulation of mRNA Splicing and Translation
OFF domain
Laskowski R;Bednarska AJ;Kramarz
PE;Loureiro S;Scheil V;Kudlek J;Holmstrup M;
2010
986
Interactions between toxic chemicals and natural
environmental factors - A meta-analysis and case studies
ON experimental
study
Lattanzi ML;Santos CB;Mudry MD;Baranao JL;
2003
486
Exposure of bovine oocytes to the endogenous metabolite OFF domain
2-methoxyestradiol during in vitro maturation inhibits early
embryonic development
Latz D;Schulze T;Manegold C;Schraub
P;Flentje M;Weber KJ;
1998
438
Combined effects of ionizing radiation and 4hydroperoxyifosfamide in vitro
OFF domain
Lau CL;
1993
697
BEHAVIOR OF EMBRYONIC CHICK HEART-CELLS IN
CULTURE .1. CELLULAR-RESPONSES TO INSULINTRANSFERRIN-SELENIUM
OFF domain
Lau YT;
2002
555
Receptor-dependent and genomic-independent actions of
estrogen in vascular protection
OFF domain
Laukkarinen J;Sand J;Aittomaki S;Porsti
I;Koobi P;Kalliovalkama J;Silvennoinen
O;Nordback I;
2002
452
Mechanism of the prorelaxing effect of thyroxine on the
sphincter of Oddi
OFF domain
Lawrence WT;Norton JA;Sporn
MB;Gorschboth C;Grotendorst GR;
1986
216
THE REVERSAL OF AN ADRIAMYCIN INDUCED HEALING
OFF domain
IMPAIRMENT WITH CHEMOATTRACTANTS AND GROWTHFACTORS
Leal-Pinto E;Teixeira A;Tran B;Hanss
B;Klotman PE;
2005
762
Presence of the nucleic acid channel in renal brush-border
membranes: allosteric modulation by extracellular calcium
OFF domain
Lebedeva II;Kuz'mina TI;Lebedev
VA;Schneider F;Alm H;Torner H;
2004
947
Prolactin-binding activity of bovine granulosa cells on
luteinization and exposure to somatotropin in vitro
OFF domain
APPENDIX A
Authors
Page 28 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Leckstrom A;Ahlner J;Grundstrom N;Axelsson
KL;
1993
881
INVOLVEMENT OF NITRIC-OXIDE AND PEPTIDES IN THE
INHIBITORY NONADRENERGIC, NONCHOLINERGIC
(NANC) RESPONSE IN BOVINE MESENTERIC-ARTERY
OFF domain
Lee JH;Landrum PF;
2006
385
Development of a multi-component damage assessment
model (MDAM) for time-dependent mixture toxicity with
toxicokinetic interactions
ON experimental
study
Lee YS;
1987
740
Adenomas, metaplastic polyps and other lesions of the
large bowel: an autopsy survey
OFF domain
Leger JM;Herrmann C;Malauzat D;Danot
G;Lombertie ER;
1984
171
ANXIOLYTICS AND MNEMONIC CAPACITY OF ELDERLY
PEOPLE - A DOUBLE-BLIND-STUDY COMPARING
TIAPRIDE AND LORAZEPAM
OFF domain
Lehenkari P;Hentunen TA;Laitala-Leinonen
T;Tuukkanen J;Vaananen HK;
1998
484
Carbonic anhydrase II plays a major role in osteoclast
differentiation and bone resorption by effecting the steady
state intracellular pH and Ca2+
OFF domain
Lehn M;Weiser WY;Engelhorn S;Gillis
S;Remold HG;
1989
289
IL-4 INHIBITS H2O2 PRODUCTION AND
ANTILEISHMANIAL CAPACITY OF HUMAN CULTURED
MONOCYTES MEDIATED BY IFN-GAMMA
OFF domain
Leibowtiz HM;Kupferman A;
1980
94
ANTI INFLAMMATORY MEDICATIONS
OFF domain
Lelievre V;Pineau N;Du J;Wen CH;Nguyen
T;Janet T;Muller JM;Waschek JA;
1998
932
Differential effects of peptide histidine isoleucine (PHI) and OFF domain
related peptides on stimulation and suppression of
neuroblastoma cell proliferation - A novel VIP-independent
action of PHI via map kinase
Levene MI;Fawer CL;Lamont RF;
1982
141
RISK-FACTORS IN THE DEVELOPMENT OF
INTRAVENTRICULAR HEMORRHAGE IN THE PRETERM
NEONATE
OFF domain
Levine MW;Shefner JM;
1981
113
DISTANCE-DEPENDENT INTERACTIONS BETWEEN THE
ROD AND THE CONE SYSTEMS IN GOLDFISH RETINA
OFF domain
Levine SA;Donowitz M;Watson AJM;Sharp
GWG;Crane JK;Weikel CS;
1991
821
CHARACTERIZATION OF THE SYNERGISTIC
INTERACTION OF ESCHERICHIA-COLI HEAT-STABLE
TOXIN AND CARBACHOL
OFF domain
Lex A;Wiesmuller KH;Jung G;Bessler WG;
1986
217
A SYNTHETIC ANALOG OF ESCHERICHIA-COLI
LIPOPROTEIN, TRIPALMITOYL PENTAPEPTIDE,
CONSTITUTES A POTENT IMMUNE ADJUVANT
OFF domain
Li Gt;Wang Hr;Li Dy;Zhao X;Liu Hj;Qu Jh;
2009
498
Enhanced Decomposition of 1, 4-Benzoquinone Ring by
DSA Electrode Under Ultraviolet Irradiation
OFF domain
Li J;Lee JM;Johnson JA;
2002
996
Microarray analysis reveals an antioxidant responsive
element-driven gene set involved in conferring protection
from an oxidative stress-induced apoptosis in IMR-32 cells
OFF domain
Li R;Shen Y;Yang LB;Lue LF;Finch C;Rogers J;
2000
601
Estrogen enhances uptake of amyloid beta-protein by
microglia derived from the human cortex
OFF domain
Liddell FDK;Armstrong BG;
2002
409
The combination of effects on lung cancer of cigarette
smoking and exposure in Quebec chrysotile miners and
millers
OFF domain
Lifschitz MD;
1986
218
PROSTAGLANDINS MAY MEDIATE CHLORIDE
CONCENTRATION GRADIENT ACROSS DOMES FORMED
BY MDCK1 CELLS
OFF domain
Lim SL;Sim MK;Loke WK;
2000
792
Acetylcholinesterase-independent action of diisopropylflurophosphate in the rat aorta
OFF domain
Lin HY;Tsay SS;
1987
857
Extracellular thermostable alpha-amylase from Bacillus
stearothermophilus Q8
OFF domain
Lin J;Chen JW;Cai XY;Qlao XL;Huang
LP;Wang D;Wang Z;
2007
584
Evolution of toxicity upon hydrolysis of fenoxaprop-p-ethyl
ON experimental
study
Lindqvist C;Lundstrom H;Okerblom
C;Akerman KEO;
1993
828
ENHANCED IL-4-MEDIATED D10.G4.1 PROLIFERATION
WITH SUBOPTIMAL CONCENTRATIONS OF ANTI-IL-4
RECEPTOR MONOCLONAL-ANTIBODIES
OFF domain
APPENDIX A
Authors
Page 29 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Liou JC;Cheng YC;Kang KH;Chu YP;Yang
CC;Chang LS;
2004
612
Both A chain and B chain of beta-bungarotoxin are
functionally involved in the facilitation of spontaneous
transmitter release in Xenopus nerve-muscle cultures
OFF domain
Lipfert L;Fisher JE;Wei N;Scafonas A;Su
Q;Yudkovitz J;Chen F;Warrier S;Birzin ET;Kim
S;Tan H;Schmidt A;Dininno F;Rohrer
SP;Hammond ML;Rodan GA;Freedman
LP;Reszka AA;
2006
539
Antagonist-induced, activation function-2-independent
estrogen receptor alpha phosphorylation
OFF domain
Liu F;Liu Ss;Liu Hl;
2007
647
Toxicities of selected ionic liquids and their mixtures to
photobacteria (Vibrio-qinghaiensis sp-Q67)
ON experimental
study
Liu QY;Chang YH;Schaffner AE;Smith
SV;Barker JL;
2002
407
Allopregnanolone activates GABA(A) receptor/Cl- channels
in a multiphasic manner in embryonic rat hippocampal
neurons
OFF domain
Liu S;Beckman JS;Ku DD;
1994
1031
PEROXYNITRITE, A PRODUCT OF SUPEROXIDE AND
NITRIC-OXIDE, PRODUCES CORONARY
VASORELAXATION IN DOGS
OFF domain
Liu SS;Song XQ;Liu HL;Zhang YH;Zhang J;
2009
1069
Combined photobacterium toxicity of herbicide mixtures
containing one insecticide
ON experimental
study
Liu X;Lukasova M;Zubakova R;Lewicka
S;Hilgenfeldt U;
2006
354
Kallidin-like peptide mediates the cardioprotective effect of
the ACE inhibitor captopril against ischaemic reperfusion
injury of rat heart
OFF domain
Lloyd-Jones KL;Kelly MM;Kubes P;
2008
916
Varying importance of soluble and membrane CD14 in
endothelial detection of lipopolysaccharide
OFF domain
Lobato EB;Beaver T;Muehlschlegel J;Kirby
DS;Klodell C;Sidi A;
2006
730
Treatment with phosphodiesterase inhibitors type III and
V: milrinone and sildenafil is an effective combination
during thromboxane-induced acute pulmonary
hypertension
OFF domain
Lock K;Janssen CR;
2002
488
Mixture toxicity of zinc, cadmium, copper, and lead to the
potworm Enchytraeus albidus
OFF domain
Lohuis J;Vanleeuwen W;Verheijden JHM;Smit
JAH;Brand A;Vanmiert A;
1988
262
GROWTH OF ESCHERICHIA-COLI IN WHOLE AND SKIM
MILK FROM ENDOTOXIN-INDUCED MASTITIC
QUARTERS - INVITRO EFFECTS OF DEFEROXAMINE,
ZINC, AND IRON SUPPLEMENTATION
OFF domain
Long SM;Reichenberg F;Lister LJ;Hankard
PK;Townsend J;Mayer P;Wright J;Holmstrup
M;Svendsen C;Spurgeon DJ;
2009
875
COMBINED CHEMICAL (FLUORANTHENE) AND DROUGHT
EFFECTS ON LUMBRICUS RUBELLUS DEMONSTRATE THE
APPLICABILITY OF THE INDEPENDENT ACTION MODEL
FOR MULTIPLE STRESSOR ASSESSMENT
ON experimental
study
Loop MS;Millican CL;Thomas SR;
1987
238
PHOTOPIC SPECTRAL SENSITIVITY OF THE CAT
OFF domain
Lounes A;Lebrihi A;Benslimane C;Lefebvre
G;Germain P;
1995
603
REGULATION OF VALINE CATABOLISM BY AMMONIUM IN
STREPTOMYCES-AMBOFACIENS, PRODUCER OF
SPIRAMYCIN
OFF domain
Loureiro S;Amorim MJB;Campos B;Rodrigues
SMG;Soares A;
2009
504
Assessing joint toxicity of chemicals in Enchytraeus albidus
(Enchytraeidae) and Porcellionides pruinosus (Isopoda)
using avoidance behaviour as an endpoint
ON experimental
study
Loureiro S;Svendsen C;Ferreira ALG;Pinheiro
C;Ribeiro F;Soares A;
2010
499
TOXICITY OF THREE BINARY MIXTURES TO DAPHNIA
MAGNA: COMPARING CHEMICAL MODES OF ACTION AND
DEVIATIONS FROM CONCEPTUAL MODELS
ON experimental
study
Lowe G;Gold GH;
1993
357
CONTRIBUTION OF THE CILIARY CYCLIC NUCLEOTIDEGATED CONDUCTANCE TO OLFACTORY TRANSDUCTION
IN THE SALAMANDER
OFF domain
Lu GH;Wang C;Wang PF;Yang CZ;
2009
432
Predicting toxicity of aromatic ternary mixtures to algae
ON experimental
study
Lu M;Hebert SG;Giebisch G;
2002
1016
Hydrolyzable ATP and PIP2 modulate the smallconductance K+ channel in apical membranes of rat
cortical-collecting duct (CCD)
OFF domain
Ludewig B;Holzmeister J;Gentile
M;Gelderblom HR;Rokos K;Becker Y;Pauli G;
1995
662
REPLICATION PATTERN OF HUMAN-IMMUNODEFICIENCY- OFF domain
VIRUS TYPE-1 IN MATURE LANGERHANS CELLS
APPENDIX A
Authors
Page 30 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Lukacs NW;Strieter RM;Evanoff HL;Burdick
MD;Kunkel SL;
1994
856
VCAM-1 INFLUENCES LYMPHOCYTE-PROLIFERATION
AND CYTOKINE PRODUCTION DURING MIXED
LYMPHOCYTE-RESPONSES
OFF domain
Lundqvist A;
1990
489
ONE-LOCUS SPOROPHYTIC S-GENE SYSTEM WITH
TRACES OF GAMETOPHYTIC POLLEN CONTROL IN
CERASTIUM-ARVENSE SSP STRICTUM
(CARYOPHYLLACEAE)
OFF domain
Lutz WK;
2009
346
Deviation from additivity in mixture toxicity: Nonlinear
dose-response and relevance for genotoxicity
ON trivial
Lutz WK;Tiedge O;Lutz RW;Stopper H;
2005
518
Different types of combination effects for the induction of
micronuclei in mouse lymphoma cells by binary mixtures
of the genotoxic agents MMS, MNU, and genistein
ON experimental
study
Lyu RM;Germano PM;Choi JK;Le SV;Pisegna
JR;
2000
989
Identification of an essential amino acid motif within the C
terminus of the pituitary adenylate cyclase-activating
polypeptide type I receptor that is critical for signal
transduction but not for receptor internalization
OFF domain
M. Goldoni;C. Johansson
2007
1139
A mathematical approach to study combined effects of
toxicants in vitro: Evaluation of the Bliss independence
criterion and the Loewe additivity model
ON review
Ma TY;Hollander D;Erickson RA;Truong
H;Nguyen H;Krugliak P;
1995
448
MECHANISM OF COLONIC PERMEATION OF INULIN - IS
RAT COLON MORE PERMEABLE THAN SMALL-INTESTINE
OFF domain
Macewan DJ;Mitchell R;Thomson FJ;Johnson
MS;
1991
950
INHIBITION OF DEPOLARIZATION-INDUCED CALCIUM
INFLUX INTO GH3 CELLS BY ARACHIDONIC-ACID - THE
INVOLVEMENT OF PROTEIN-KINASE-C
OFF domain
Machado SG;Bailey KR;
1985
192
ASSESSMENT OF INTERACTION BETWEEN CARCINOGENS OFF domain
IN LONG-TERM FACTORIALLY DESIGNED ANIMALEXPERIMENTS
Macharia R;Patel K;Otto WR;McKinnell
IW;Christ B;
2004
1052
Decamethonium bromide-mediated inhibition of embryonic
muscle development
OFF domain
Maciejewska B;Kopcewicz J;
2002
899
Inhibitory effect of methyl jasmonate on flowering and
elongation growth in Pharbitis nil
OFF domain
Mack CM;McGivern RF;Hyde LA;Denenberg
VH;
1996
1103
Absence of postnatal testosterone fails to demasculinize
the male rat's corpus callosum
OFF domain
Maclean CJ;Sham PC;Kendler KS;
1993
674
JOINT LINKAGE OF MULTIPLE LOCI FOR A COMPLEX
DISORDER
OFF domain
MacMillan CM;
2006
877
Active conscience or administrative vanguard?: The
Commissioner of Official Languages as an agent of change
OFF domain
Mager EM;Wintz H;Vulpe CD;Brix KV;Grosell
M;
2008
537
Toxicogenomics of water chemistry influence on chronic
lead exposure to the fathead minnow (Pimephales
promelas)
ON experimental
study
Magnani M;Stocchi V;Serafini G;Chiarantini L;
1988
263
THE INTERACTION OF PHOSPHORYLATED SUGARS WITH
HUMAN HEXOKINASE-I
OFF domain
Magnusson L;Rebagliati M;Buchser E;
1993
852
INCREASED ARTERIAL-PRESSURE DURING SURGERY
WITH PNEUMATIC TOURNIQUET - MECHANISM
UNAFFECTED BY RENIN
OFF domain
Mahmoud ME;Shimizu Y;Shiina T;Nikami
H;Dosoky RM;Ahmed MM;Takewaki T;
2007
650
Involvement of a capsaicin-sensitive TRPV1-independent
mechanism in lipopolysaccharide-induced fever in chickens
OFF domain
Majury AL;Shewen PE;
1991
746
PRELIMINARY INVESTIGATION OF THE MECHANISM OF
INHIBITION OF BOVINE LYMPHOCYTE-PROLIFERATION
BY PASTEURELLA-HAEMOLYTICA A1 LEUKOTOXIN
OFF domain
Maki T;Satomi S;Gotoh M;Monaco AP;
1986
219
CONTRA-IL 2 - A SUPPRESSOR LYMPHOKINE THAT
INHIBITS IL-2 ACTIVITY
OFF domain
Malejczyk J;Kaminski MJ;Malejczyk
M;Majewski S;
1985
193
NATURAL CELL-MEDIATED CYTO-TOXIC ACTIVITY
AGAINST ISOLATED CHONDROCYTES IN THE MOUSE
OFF domain
Malinow MR;McLaughlin P;Stafford
C;Livingston AL;Kohler GO;Cheeke PR;
1979
80
COMPARATIVE EFFECTS OF ALFALFA SAPONINS AND
ALFALFA FIBER ON CHOLESTEROL ABSORPTION IN RATS
OFF domain
APPENDIX A
Authors
Page 31 of 66
PubYear
ID
Title
CFT/EFSA/PPR/2010/02
Relevance
Malygin EG;Evdokimov AA;Zinoviev
VV;Ovechkina LG;Lindstrom WM;Reich
NO;Schlagman SL;Hattman S;
2001
926
A dual role for substrate S-adenosyl-L-methionine in the
OFF domain
methylation reaction with bacteriophage T4 Dam DNA- N6adenine -methyltransferase
Mann DL;Martin P;Hamlingreen G;Nalewaik
R;Blattner W;
1994
508
VIRUS PRODUCTION AND SPONTANEOUS CELLPROLIFERATION IN HTLV-I-INFECTED LYMPHOCYTES
OFF domain
Manos PJ;Goldthwaite J;
1975
38
KINETIC-ANALYSIS OF EFFECTS OF GIBBERELLIC-ACID,
ZEATIN, AND ABSCISIC-ACID ON LEAF TISSUE
SENESCENCE IN RUMEX
OFF domain
Manucha W;Oliveros L;Carrizo L;Seltzer
A;Valles P;
2004
443
Losartan modulation on NOS isoforms and COX-2
expression in early renal fibrogenesis in unilateral
obstruction
OFF domain
Manzo S;Buono S;Cremisini C;
2010
851
Cadmium, lead and their mixtures with copper:
Paracentrotus lividus embryotoxicity assessment,
prediction, and offspring quality evaluation
ON experimental
study
Mao ZQ;Liu LQ;Zhang RS;Li XH;
2007
456
Lithium reduces FoxO(3)a transcriptional activity by
decreasing its intracellular content
OFF domain
Marano G;Palazzesi S;Fadda A;Vergari
A;Ferrari AU;
2002
714
Attenuation of aortic banding-induced cardiac hypertrophy
by propranolol is independent of beta-adrenoceptor
blockade
OFF domain
Marcelle R;
1971
16
EFFECTS OF GROWTH RETARDANTS B-9 AND CCC ON
GA3-INDUCED THORN FORMATION BY YOUNG PEAR
TREES
OFF domain
Marchi B;Burlando B;Panfoli I;Viarengo A;
2000
604
Interference of heavy metal cations with fluorescent Ca2+
probes does not affect Ca2+ measurements in living cells
OFF domain
Marcoux J;Deziel E;Villemur R;Lepine
F;Bisaillon JG;Beaudet R;
2000
381
Optimization of high-molecular-weight polycyclic aromatic
OFF domain
hydrocarbons' degradation in a two-liquid-phase bioreactor
Mariage TV;
2001
1050
Features of an interactive writing discourse:
Conversational involvement, conventional knowledge, and
internalization in 'morning message'
OFF domain
Markostamos K;Andrikopoulou M;
1992
738
TEMPOROMANDIBULAR JOINT DYSFUNCTION
ORTHODONTIC APPROACH
OFF domain
Martin HL;Svendsen C;Lister LJ;Gomez-Eyles
JL;Spurgeon DJ;
2009
522
MEASUREMENT AND MODELING OF THE TOXICITY OF
BINARY MIXTURES IN THE NEMATODE
CAENORHABDITIS ELEGANS-A TEST OF INDEPENDENT
ACTION
ON experimental
study
Martin SP;
1988
264
CONFORMAL FIELD-THEORY REPRESENTATION AND
BACKGROUND-INDEPENDENT ACTION FOR
SUPERSTRING FIELD-THEORY
OFF domain
Marzella L;Yu QH;Mergner W;Trump BF;
1984
172
UNBUFFERED OSMIUM STAINING OF CELL
ORGANELLES - ALTERATIONS INDUCED BY CELL INJURY
OFF domain
Masterson TJ;Hartwig WC;
1998
814
Degrees of sexual dimorphism in Cebus and other new
world monkeys
OFF domain
Matsuda H;Hayashi K;Wakino S;Kubota
E;Honda M;Tokuyama H;Takamatsu
I;Tatematsu S;Saruta T;
2004
411
Role of endothelium-derived hyperpolarizing factor in ACE
inhibitor-induced renal vasodilation in vivo
OFF domain
Mattano SS;Hollenberg PF;
1994
1011
THE EFFECT OF CYTOSOL LIVER MICROSOMAL
METABOLIC-ACTIVATION AND DEMETHYLATION OF NNITROSODIMETHYLAMINE
OFF domain
Matzke M;Stolte S;Boschen A;Filser J;
2008
1087
Mixture effects and predictability of combination effects of
imidazolium based ionic liquids as well as imidazolium
based ionic liquids and cadmium on terrestrial plants
(Triticum aestivum) and limnic green algae (Scenedesmus
vacuolatus)
ON experimental
study
May ME;Aftring RP;Buse MG;
1980
95
MECHANISM OF THE STIMULATION OF BRANCHEDCHAIN OXOACID OXIDATION IN LIVER BY CARNITINE
OFF domain
Mayer MP;Schroder H;Rudiger S;Paal
K;Laufen T;Bukau B;
2000
838
Multistep mechanism of substrate binding determines
chaperone activity of Hsp70
OFF domain
APPENDIX A
Authors
Page 32 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Mazei-Robison MS;Blakely RD;
2005
694
Expression studies of naturally occurring human dopamine
transporter variants identifies a novel state of transporter
inactivation associated with Val382Ala
OFF domain
McClellan K;Altenburger R;Schmitt-Jansen M;
2008
362
Pollution-induced community tolerance as a measure of
species interaction in toxicity assessment
ON method dev.
McCulloch CR;Kuwahara A;Condon CD;Cooke
HJ;
1987
239
NEUROPEPTIDE MODIFICATION OF CHLORIDE
SECRETION IN GUINEA-PIG DISTAL COLON
OFF domain
McDonald MJ;Michalski LA;Turci SM;Guillette
RA;Jue DL;Johnson MH;Moopenn WF;
1990
309
STRUCTURAL, FUNCTIONAL, AND SUBUNIT ASSEMBLY
PROPERTIES OF HEMOGLOBIN ATTLEBORO ALPHA-138
(H21) SER- PRO , A VARIANT POSSESSING A SITE
MUTATION AT A CRITICAL C-TERMINAL RESIDUE
OFF domain
McDonough SI;Cseresnyes Z;Schneider MF;
2000
595
Origin sites of calcium release and calcium oscillations in
frog sympathetic neurons
OFF domain
McDowell I;Newell C;Rosser W;
1990
501
A follow-up study of patients advised to obtain influenza
immunizations
OFF domain
McLaughlin CW;Peart D;Purves RD;Carre
DA;Peterson-Yantorno K;Mitchell
CH;Macknight ADC;Civan MM;
2001
868
Timolol may inhibit aqueous humor secretion by cAMPindependent action on ciliary epithelial cells
OFF domain
McNally NJ;Deronde J;Hinchliffe M;
1985
194
SURVIVAL OF V79 CELLS FOLLOWING SIMULTANEOUS
IRRADIATION WITH X-RAYS AND NEUTRONS IN AIR OR
HYPOXIA
OFF domain
Meachem SJ;Wreford NG;Stanton
PG;Robertson DM;McLachlan RI;
1998
1009
Follicle-stimulating hormone is required for the initial
phase of spermatogenic restoration in adult rats following
gonadotropin suppression
OFF domain
Medvedev AE;Gorkin VZ;
1992
623
BIOGENIC-AMINES AND MONOAMINE OXIDASES IN THE
REGULATION OF ACTIVITIES OF MEMBRANE-BOUND
MITOCHONDRIAL-ENZYMES
OFF domain
Mehlenbacher SA;
1997
755
Revised dominance hierarchy for S-alleles in Corylus
avellana L
OFF domain
Mehlenbacher SA;Thompson MM;
1988
265
DOMINANCE RELATIONSHIPS AMONG S-ALLELES IN
CORYLUS-AVELLANA L
OFF domain
Mei JB;Reineccius GA;Knighton WB;Grimsrud
EP;
2004
998
Influence of strawberry yogurt composition on aroma
release
OFF domain
Melfi R;Nusca A;Patti G;Di Sciascio G;
2010
665
Statins and their role in pre-percutaneous coronary
intervention
OFF domain
Mellgren RL;Song K;Mericle MT;
1993
669
M-CALPAIN REQUIRES DNA FOR ACTIVITY ON NUCLEAR
PROTEINS AT LOW CALCIUM CONCENTRATIONS
OFF domain
Meltzer D;Burger PA;
1983
155
STIMULUS DISCRIMINABILITY AND CONDITIONINGHISTORY EFFECTS ON RESPONSE SUMMATION
OFF domain
Meltzer D;Freeman BJ;
1971
17
MAINTENANCE OF RESPONSE SUMMATION UNDER
CONDITIONS OF MINIMUM STIMULUS INTENSITY
OFF domain
Meltzer D;Hamm RJ;
1976
50
RESPONSE SUMMATION IN PIGEON
OFF domain
Meltzer D;Masaki MA;
1972
24
STIMULUS PARAMETERS PRODUCING RESPONSE
SUMMATION
OFF domain
Meltzer D;Niebuhr B;
1981
114
RESPONSE SUMMATION AND NONCONTINGENT
REINFORCEMENT
OFF domain
Meltzer D;Niebuhr BR;
1974
30
ADDITIVE AND SUPPRESSIVE RESPONSE SUMMATION
WITH A CHAIN SCHEDULE
OFF domain
Meltzer D;Niebuhr BR;Hamm RJ;
1979
81
RESPONSE SUMMATION WITH DISCRIMINATIVE STIMULI OFF domain
CONTROLLING RESPONDING ON SEPARATE
MANIPULANDA
Mendelsohn J;Trowbridge I;Castagnola J;
1983
156
INHIBITION OF HUMAN-LYMPHOCYTE PROLIFERATION
BY MONOCLONAL-ANTIBODY TO TRANSFERRIN
RECEPTOR
OFF domain
APPENDIX A
Authors
Page 33 of 66
PubYear
ID
Title
CFT/EFSA/PPR/2010/02
Relevance
Menegaz D;Royer C;Rosso A;de Souza
AZP;dos Santos ARS;Silva F;
2010
412
Rapid stimulatory effect of thyroxine on plasma membrane OFF domain
transport systems: Calcium uptake and neutral amino acid
accumulation in immature rat testis
Meng WZ;Nicholson AW;
2008
677
Heterodimer-based analysis of subunit and domain
contributions to double-stranded RNA processing by
Escherichia coli RNase III in vitro
OFF domain
Merino-Garcia D;Kusk KO;Christensen ER;
2003
1027
Joint toxicity of similarly and dissimilarly acting chemicals
to Daphnia magna at different response levels
ON experimental
study
Meynell GG;
1957
4
THE APPLICABILITY OF THE HYPOTHESIS OF
INDEPENDENT ACTION TO FATAL INFECTIONS IN MICE
GIVEN SALMONELLA-TYPHIMURIUM BY MOUTH
OFF domain
Miazzo R;Peralta MF;Magnoli C;Salvano
M;Ferrero S;Chiacchiera SM;Carvalho
ECQ;Rosa CAR;Dalcero A;
2005
818
Efficacy of sodium bentonite as a detoxifier of broiler feed
contaminated with aflatoxin and fumonisin
OFF domain
Michalk A;Stricker S;Becker J;Rupps
R;Pantzar T;Miertus J;Botta G;Naretto
VG;Janetzki C;Yaqoob N;Ott CE;Seelow
D;Wieczorek D;Fiebig B;Wirth B;Hoopmann
M;Walther M;Koerber F;Blankenburg
M;Mundlos S;Heller R;Hoffmann K;
2008
586
Acetylcholine receptor pathway mutations explain various
fetal akinesia deformation sequence disorders
OFF domain
Middeke M;
2001
479
Britain's global military role, conventional defence and
Anglo-American interdependence after Nassau
OFF domain
Mikkelsen A;Bertelsen G;Skibsted LH;
1991
516
POLYPHOSPHATES AS ANTIOXIDANTS IN FROZEN BEEF
PATTIES - LIPID OXIDATION AND COLOR QUALITY
DURING RETAIL DISPLAY
OFF domain
Milev S;Bosshard HR;Jelesarov I;
2005
898
Enthalpic and entropic effects of salt and polyol osmolytes
on site-specific protein-DNA association: The integrase
Tn916-DNA complex
OFF domain
Miller CL;Baker CC;
1979
82
CHANGES IN LYMPHOCYTE ACTIVITY AFTER THERMAL
INJURY - ROLE OF SUPPRESSOR CELLS
OFF domain
Miller L;
1973
27
COMPOUNDING OF DISCRIMINATIVE STIMULI THAT
MAINTAIN RESPONDING ON SEPARATE RESPONSE
LEVERS
OFF domain
Miller L;Price RD;
1971
18
STIMULUS COMPOUNDING AND RESPONSE SUMMATION
WITH AN INSTRUMENTAL RUNNING RESPONSE
OFF domain
Miller RR;Grahame NJ;Hallam SC;
1990
310
SUMMATION OF RESPONDING TO CSS AND AN
EXCITATORY TEST CONTEXT
OFF domain
Millier PJ;Beale IL;
1977
57
RESPONSE SUMMATION TO A COMPOUND STIMULUS IN
A CONTEXT OF CHOICE
OFF domain
Minang JT;Arestrom I;Zuber B;Jonsson
G;Troye-Blomberg M;Ahlborg N;
2006
1124
Nickel-induced IL-10 down-regulates Th1- but not Th2type cytokine responses to the contact allergen nickel
OFF domain
Mine T;Kojima I;Ogata E;
1988
266
EVIDENCE OF CYCLIC AMP-INDEPENDENT ACTION OF
GLUCAGON ON CALCIUM MOBILIZATION IN RAT
HEPATOCYTES
OFF domain
Minotti G;Ronchi R;Salvatorelli E;Menna
P;Cairo G;
2001
492
Doxorubicin irreversibly inactivates iron regulatory proteins
1 and 2 in cardiomyocytes: Evidence for distinct metabolic
pathways and implications for iron-mediated cardiotoxicity
of antitumor therapy
OFF domain
Mio T;Liu XD;Toews ML;Adachi Y;Romberger
DJ;Spurzem JR;Rennard SI;
2001
478
Bradykinin augments fibroblast-mediated contraction of
released collagen gels
OFF domain
Mio Y;Pravdic D;Bosnjak ZJ;Bienengraeber M;
2009
347
IMMEDIATE PROTECTIVE MECHANISMS IN
POSTCONDITIONING
OFF domain
Mirzoyan GI;
1984
981
INDUCED MUTABILITY OF CREPIS-CAPILLARIS
CHROMOSOMES DURING SEED STORAGE AND
MODIFICATION OF DNA SYNTHESIS
OFF domain
Mishima Y;Ohyama Y;Shibata T;Seto H;Hatae
S;
1994
1104
Inhibitory action of Kojic acid on melanogenesis and its
therapeutic effect for various human hyper-pigmentation
disorders
OFF domain
APPENDIX A
Authors
Page 34 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Missiaen L;DeSmedt H;Parys JB;Sienaert
I;Sipma H;Vanlingen S;Casteels R;
1997
629
Slow kinetics of inositol 1,4,5-trisphosphate-induced Ca2+
release: Is the release 'quantal' or 'non-quantal'?
OFF domain
Mitchell I;Hayward S;Deshpande N;
1988
267
RECEPTOR-INDEPENDENT ACTION OF HORMONES ON
THE ACTIVITIES OF CERTAIN ENZYMES IN HUMANBREAST CANCER CELL-LINE BT-20
OFF domain
Mittag TW;Hammond BR;Eakins
KE;Bhattacherjee P;
1985
195
OCULAR RESPONSES TO SUPEROXIDE GENERATED BY
INTRAOCULAR INJECTION OF XANTHINE-OXIDASE
OFF domain
Miura KI;Kodama Y;Shimotohna K;Fukui
T;Ikehara M;Nakagawa I;Hata T;
1979
83
INHIBITORY EFFECT OF METHYLATED DERIVATIVES OF
OFF domain
GUANYLIC ACID FOR PROTEIN-SYNTHESIS WITH
REFERENCE TO THE FUNCTIONAL STRUCTURE OF THE 5'CAP IN VIRAL MESSENGER-RNA
Miura Y;
1984
173
Assistance of a patient with subarachnoid hemorrhage for
independent action at mealtime--nursing a 20-year-old
man with a paralyzed left hand and fitted with an artificial
right hand
OFF domain
Miyake Y;Hirose T;Hara A;
1983
157
ELECTROPHYSIOLOGIC TESTING OF VISUAL FUNCTIONS
FOR VITRECTOMY CANDIDATES .1. RESULTS IN EYES
WITH KNOWN FUNDUS DISEASES
OFF domain
Moleiro FC;Andreo MA;dos Santos RD;Moraes
TD;Rodrigues CM;Carli CBD;Lopes
FCM;Pellizzon CH;Carlos IZ;Bauab TM;Vilegas
W;Hiruma-Lima CA;
2009
541
Mouriri elliptica: Validation of gastroprotective, healing and OFF domain
anti-Helicobacter pylori effects
Molinari JF;Campo C;Shakir S;Ahmed T;
1998
1086
Inhibition of antigen-induced airway hyperresponsiveness
by ultralow molecular-weight heparin
OFF domain
Moniuszko-Jakoniuk J;Jurczuk M;Brzoska MM;
2007
796
Evaluation of glutathione-related enzyme activities in the
liver and kidney of rats exposed to lead and ethanol
OFF domain
Moniuszko-Jakoniuk J;Jurczuk M;GalazynSidorczuk M;Brzoska MM;
2003
895
Lead turnover and changes in the body status of chosen
micro- and macroelements in rats exposed to lead and
ethanol
OFF domain
Montesinos E;Bonaterra A;
1996
1078
Dose-response models in biological control of plant
pathogens: An empirical verification
ON experimental
study
Moopenn WF;McGuffey JE;Jue DL;Johnson
MH;Schum T;
1985
196
HEMOGLOBIN NEW MEXICO - BETA-100 (G2) PRO- ARG A VARIANT HEMOGLOBIN ASSOCIATED WITH
ERYTHROCYTOSIS
OFF domain
Mora-Gutierrez A;Farrell HM;
2000
711
Sugar-casein interaction in deuterated solutions of bovine
and caprine casein as determined by oxygen-17 and
carbon-13 nuclear magnetic resonance: A case of
preferential interactions
OFF domain
Moretto MM;Lawlor EM;Khan IA;
2010
368
Lack of Interleukin-12 in p40-Deficient Mice Leads to Poor
CD8(+) T-Cell Immunity against Encephalitozoon cuniculi
Infection
OFF domain
Morganroth ML;Till GO;Kunkel RG;Ward PA;
1986
220
COMPLEMENT AND NEUTROPHIL-MEDIATED INJURY OF
PERFUSED RAT LUNGS
OFF domain
Morisawa K;
1989
599
EFFECTS OF GROWTH HORMONE AND THYROID
HORMONE ON BRAIN DEVELOPMENT
OFF domain
Morowitz HJ;Morowitz LS;
1977
58
RADIATION INACTIVATION OF OXIDATIVEPHOSPHORYLATION
OFF domain
Mosse HL;
1953
2
Is there an Ishmael complex?
OFF domain
Moustoukas N;Browder W;Gleason C;Diluzio
N;Nichols RL;
1985
197
ADVERSE EFFECT OF SPLENECTOMY IN EXPERIMENTAL
PERITONITIS
OFF domain
Moxon ER;Murphy PA;
1978
69
HEMOPHILUS-INFLUENZAE BACTEREMIA AND
MENINGITIS RESULTING FROM SURVIVAL OF A SINGLE
ORGANISM
OFF domain
Mras S;Sperelakis N;
1981
115
BEPRIDIL (CERM-1978) BLOCKADE OF ACTIONPOTENTIALS IN CULTURED RAT AORTIC SMOOTHMUSCLE CELLS
OFF domain
APPENDIX A
Authors
Page 35 of 66
PubYear
ID
Title
CFT/EFSA/PPR/2010/02
Relevance
Mu XY;LeBlanc GA;
2004
530
Synergistic interaction of endocrine-disrupting chemicals:
ON experimental
Model development using an ecdysone receptor antagonist study
and a hormone synthesis inhibitor
Munkegaard M;Abbaspoor M;Cedergreen N;
2008
687
Organophosphorous insecticides as herbicide synergists on ON experimental
the green algae Pseudokirchneriella subcapitata and the
study
aquatic plant Lemna minor
Murado MA;Vazquez JA;Rial D;Beiras R;
2011
816
Dose-response modelling with two agents: Application to
the bioassay of oil and shoreline cleaning agents
ON experimental
study
Muraoka S;
1991
543
CYTOTOXIC LYMPHOCYTE-T PRECURSOR CELLS
SPECIFIC FOR THE MAJOR HISTOCOMPATIBILITY
COMPLEX CLASS-I-LIKE ANTIGEN, QA-2, REQUIRE CD4+
T-CELLS TO BECOME PRIMED INVIVO AND TO
DIFFERENTIATE INTO EFFECTOR-CELLS INVITRO
OFF domain
Mwense M;Wang XZ;Buontempo FV;Horan
N;Young A;Osborn D;
2004
510
Prediction of noninteractive mixture toxicity of organic
compounds based on a fuzzy set method
ON method dev.
Mzali H;Guichard C;Lavau M;Plas C;
1997
970
Time-dependent effects of insulin on lipid synthesis in
cultured fetal rat hepatocytes: A comparison between
lipogenesis and glycogenesis
OFF domain
Na Lu;Zhen-jia Liu;Zheng Yan;Nan Wang
2010
1145
Three different drug interaction mathematical models used ON experimental
to evaluate the cytotoxicity of SAHA and arsenic trioxide in study
combination
Nagao M;Sugimori M;Nakafuku M;
2007
760
Cross talk between Notch and growth factor/cytokine
signaling pathways in neural stem cells
OFF domain
Nagasawa S;
1976
972
PRESUMPTION OF DOSAGE RESPONSE CURVE OBTAINED
BY THE TREATMENT OF CHEMO STERILANT FOR BOTH
SEXES OF THE MEXICAN BEAN WEEVIL ZABROTESSUBFASCIATUS COLEOPTERA BRUCHIDAE
OFF domain
Nagell CF;Wettergren A;Pedersen
JF;Mortensen D;Holst JJ;
2004
458
Glucagon-like peptide-2 inhibits antral emptying in man,
but is not as potent as glucagon-like peptide-1
OFF domain
Nakajima A;Yamada K;Zou LB;Yan YJ;Mizuno
M;Nabeshima T;
2002
741
Interleukin-6 protects PC12 cells from 4-hydroxynonenalinduced cytotoxicity by increasing intracellular glutathione
levels
OFF domain
Nakamuta H;Koida M;Ogawa Y;Orlowski RC;
1987
240
CENTRALLY AND PERIPHERALLY INDUCED ANORECTIC
ACTIONS OF SALMON-CALCITONIN (SCT) IN RATS SEPARATION IN ITS NOVEL DERIVATIVE GLY8 -SCT
OFF domain
Nandanan E;Jang SY;Moro S;Kim HO;Siddiqui
MA;Russ P;Marquez VE;Busson R;Herdewijn
P;Harden TK;Boyer JL;Jacobson KA;
2000
1018
Synthesis, biological activity, and molecular modeling of
ribose-modified deoxyadenosine bisphosphate analogues
as P2Y(1) receptor ligands
OFF domain
Nangle LA;Motta CM;Schimmel P;
2006
414
Global effects of mistranslation from an editing defect in
mammalian cells
OFF domain
Naylor J;Al-Shawaf E;McKeown L;Manna
PT;Porter KE;O'Regan D;Muraki K;Beech DJ;
2011
1072
TRPC5 Channel Sensitivities to Antioxidants and
Hydroxylated Stilbenes
OFF domain
Nazare JA;Normand S;Triantafyllou AO;de la
Perriere AB;Desage M;Laville M;
2009
512
Modulation of the postprandial phase by beta-glucan in
OFF domain
overweight subjects: Effects on glucose and insulin kinetics
Neulen J;Williams RF;Breckwoldt M;Chwalisz
K;Baulieu EE;Hodgen GD;
1996
766
Non-competitive anti-oestrogenic actions of progesterone
antagonists in primate endometrium: Enhancement of
oestrogen and progesterone receptors with blockade of
post-receptor proliferative mechanisms
OFF domain
Newcomb LL;Diderich JA;Slattery
MG;Heideman W;
2003
710
Glucose regulation of Saccharomyces cerevisiae cell cycle
genes
OFF domain
Newton J;Hunt J;Stirling J;
1996
441
Human resource management in general practice: Survey
of current practice
OFF domain
Ngo FQH;Blakely EA;Tobias CA;
1981
116
SEQUENTIAL EXPOSURES OF MAMMALIAN-CELLS TO
LOW-LET AND HIGH-LET RADIATIONS .1. LETHAL
EFFECTS FOLLOWING X-RAY AND NEON-ION
IRRADIATION
OFF domain
Niki I;Niwa T;Fukasawa T;Hidaka H;
1998
331
A K-ATP-independent action of glucose may be granule
translocation by intracellular Ca2+ mobilization
OFF domain
APPENDIX A
Authors
Page 36 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Noguchi T;Sugiasaki T;Tsukada Y;
1985
198
MICROCEPHALIC CEREBRUM WITH HYPOMYELINATION
IN THE GROWTH HORMONE-DEFICIENT MOUSE (LIT)
OFF domain
Noguchi T;Sugisaki T;
1985
199
ABNORMAL NEURONAL GROWTH IN THE LITTLE (LIT)
CEREBRUM
OFF domain
Nordquist L;Lai EY;Sjoquist M;Patzak
A;Persson AEG;
2008
718
Proinsulin C-peptide constricts glomerular afferent
arterioles in diabetic mice. A potential renoprotective
mechanism
OFF domain
Norgaard KB;Cedergreen N;
2010
918
Pesticide cocktails can interact synergistically on aquatic
crustaceans
ON experimental
study
Norwood WP;Borgmann U;Dixon DG;Wallace
A;
2003
367
Effects of metal mixtures on aquatic biota: A review of
observations and methods
ON review
Ntalli NG;Ferrari F;Giannakou I;MenkissogluSpiroudi U;
2011
450
Synergistic and antagonistic interactions of terpenes
against Meloidogyne incognita and the nematicidal activity
of essential oils from seven plants indigenous to Greece
ON experimental
study
Nusca A;Melfi R;Patti G;Sciascio GD;
2010
824
Statin loading before percutaneous coronary intervention:
proposed mechanisms and applications
OFF domain
Nussenzweig MC;Steinman RM;Gutchinov
B;Cohn ZA;
1980
96
DENDRITIC CELLS ARE ACCESSORY CELLS FOR THE
DEVELOPMENT OF ANTI-TRINITROPHENYL CYTO-TOXIC
T-LYMPHOCYTES
OFF domain
Oenfelt A;Magnusson J;Wiken M;
1993
812
Ageing before mating and quinacrine ameliorate the
expression of abnormal oocyte (abo) in homozygous
Drosophila melanogaster females
OFF domain
Oertland JJ;Ahmadi N;
1975
949
The effect of Systox on ionic fluxes in excised barley roots
ON trivial
Ofosu FA;Blajchman MA;Modi G;Cerskus
AL;Hirsh J;
1981
117
ACTIVATION OF FACTOR-X AND PROTHROMBIN IN
ANTITHROMBIN-III DEPLETED PLASMA - THE EFFECTS
OF HEPARIN
OFF domain
Ogura T;Imanishi S;Shibamoto T;
2001
911
Activation of background membrane conductance by the
tyrosine kinase inhibitor tyrphostin A23 and its inactive
analog tyrphostin A1 in guinea pig ventricular myocytes
OFF domain
Oh Y;
1998
935
IGF-independent regulation of breast cancer growth by
IGF binding proteins
OFF domain
Oh Y;Rosenfeld RG;
1998
954
Old and new concepts of insulin-like growth factor binding
proteins
OFF domain
Oh YM;Muller HL;Lamson G;Rosenfeld RG;
1993
538
INSULIN-LIKE GROWTH-FACTOR (IGF)-INDEPENDENT
ACTION OF IGF-BINDING PROTEIN-3 IN HS578T HUMAN
BREAST-CANCER CELLS - CELL-SURFACE BINDING AND
GROWTH-INHIBITION
OFF domain
Ohlsson A;Cedergreen N;Oskarsson A;Ulleras
E;
2010
545
Mixture effects of imidazole fungicides on cortisol and
aldosterone secretion in human adrenocortical H295R cells
ON experimental
study
Ohlsson A;Ulleras E;Cedergreen N;Oskarsson
A;
2010
581
Mixture effects of dietary flavonoids on steroid hormone
synthesis in the human adrenocortical H295R cell line
ON experimental
study
Ohnishi T;Nakamura O;Arakaki N;Daikuhara
Y;
1997
786
Effect of phosphorylated rat fetuin on the growth of
hepatocytes in primary culture in the presence of human
hepatocyte-growth factor - Evidence that phosphorylated
fetuin is a natural modulator of hepatocyte-growth factor
OFF domain
Ohno-Matsui K;Yoshida T;Uetama
T;Mochizuki M;Morita I;
2003
1113
Vascular endothelial growth factor upregulates pigment
epithelium-derived factor expression via VEGFR-1 in
human retinal pigment epithelial cells
OFF domain
Okopien B;Krysiak R;Kowalski J;Madej
A;Belowski D;Zielinski M;Labuzek K;Herman
ZS;
2004
850
The effect of statins and fibrates on interferon-gamma and OFF domain
interleukin-2 release in patients with primary type II
dyslipidemia
Olenik TJ;Cheng SL;
1994
661
LAND-DEVELOPMENT REGULATIONS - ROADBLOCK TO
AFFORDABLE HOUSING
OFF domain
Olesen KH;
1971
19
NATRIURETIC EFFECT ADDITION OF QUINETHAZONE
AND FUROSEMIDE IN CONGESTIVE HEART FAILURE PERMUTATION TRIAL TEST
OFF domain
APPENDIX A
Authors
Page 37 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Olesen KH;Sigurd B;
1971
20
SUPRA-ADDITIVE NATRIURETIC EFFECT ADDITION OF
QUINETHAZONE OR BENDROFLUMETHIAZIDE DURING
LONG-TERM TREATMENT WITH FUROSEMIDE AND
SPIRONOLACTONE - PERMUTATION TRIAL TESTS IN
PATIENTS WITH CONGESTIVE HEART FAILURE
OFF domain
Olmstead AW;LeBlanc GA;
2003
660
Insecticidal juvenile hormone analogs stimulate the
production of male offspring in the crustacean Daphnia
magna
OFF domain
Olmstead AW;LeBlanc GA;
2005
617
Toxicity assessment of environmentally relevant pollutant
mixtures using a heuristic model
ON experimental
study
Olmstead AW;LeBlanc GA;
2005
758
Joint action of polycyclic aromatic hydrocarbons: Predictive ON experimental
modeling of sublethal toxicity
study
Olsnes C;Heimdal JH;Kross K;Olofsson
J;Aarstad HJ;
2002
472
Mechanisms for monocyte activation in co-culture with
autologous tumor spheroids
OFF domain
Omoto CK;Brokaw CJ;
1982
142
STRUCTURE AND BEHAVIOR OF THE SPERM TERMINAL
FILAMENT
OFF domain
Oneill GJ;Pearson BA;Davies DAL;
1975
39
INVITRO CYTOTOXICITY OF ANTI-THETA(THY-1)
ANTIBODIES COMBINED WITH CHLORAMBUCIL
OFF domain
Orias E;Gartner TK;
1966
7
SUPPRESSION OF AMBER AND OCHRE R2 MUTANTS OF
BACTERIOPHAGE T4 BY STREPTOMY IN
OFF domain
Ortmann O;Tomic M;Weiss JM;Diedrich
K;Stojilkovic SS;
1998
420
Dual action of androgen on calcium signaling and
luteinizing hormone secretion in pituitary gonadotrophs
OFF domain
Oshaughnessy PJ;Pearce S;Mannan MA;
1990
311
EFFECT OF HIGH-DENSITY-LIPOPROTEIN ON BOVINE
GRANULOSA-CELLS - PROGESTERONE PRODUCTION IN
NEWLY ISOLATED CELLS AND DURING CELL-CULTURE
OFF domain
Otani H;Das DK;
1988
268
ALPHA-1-ADRENOCEPTOR MEDIATED
PHOSPHOINOSITIDE BREAKDOWN AND INOTROPIC
RESPONSE IN RAT LEFT-VENTRICULAR PAPILLARYMUSCLES
OFF domain
Overgaard J;
1980
97
EFFECT OF MISONIDAZOLE AND HYPERTHERMIA ON THE OFF domain
RADIOSENSITIVITY OF A C3H MOUSE MAMMARYCARCINOMA AND ITS SURROUNDING NORMAL TISSUE
Overgaard J;Gonzalez DG;Hulshof
M;Arcangeli G;Dahl O;Mella O;Bentzen SM;
1996
919
Hyperthermia as an adjuvant to radiation therapy of
recurrent or metastatic malignant melanoma. A
multicentre randomized trial by the European Society for
Hyperthermic Oncology
OFF domain
Overmyer JP;Smith PF;Kellock KA;Kwon
JW;Armbrust KL;
2010
811
Assessment of the Toxicological Interaction of Sertraline
with Cholinesterase Inhibiting Insecticides in Aquatic
Insects Using the Black Fly, Simulium vittatum IS-7
ON experimental
study
Owens E;
2007
1102
Nonbiologic objects as actors
OFF domain
Paavonen T;
1985
200
GLUCOCORTICOIDS ENHANCE THE INVITRO IG
SYNTHESIS OF POKEWEED MITOGEN-STIMULATED
HUMAN B-CELLS BY INHIBITING THE SUPPRESSIVE
EFFECT OF T8+ T-CELLS
OFF domain
Paller AS;Arnsmeier SL;Alvarezfranco
M;Bremer EG;
1993
559
GANGLIOSIDE G(M3) INHIBITS THE PROLIFERATION OF
CULTURED KERATINOCYTES
OFF domain
Palombella VJ;Vilcek J;
1989
290
MITOGENIC AND CYTO-TOXIC ACTIONS OF TUMOR
NECROSIS FACTOR IN BALB/C 3T3 CELLS - ROLE OF
PHOSPHOLIPASE ACTIVATION
OFF domain
Pandey KK;
1981
118
EVOLUTION OF UNILATERAL INCOMPATIBILITY IN
FLOWERING PLANTS - FURTHER EVIDENCE IN FAVOR OF
TWIN SPECIFICITIES CONTROLLING INTRASPECIFIC
AND INTERSPECIFIC INCOMPATIBILITY
OFF domain
PapeLindstrom PA;Lydy MJ;
1997
393
Synergistic toxicity of atrazine and organophosphate
insecticides contravenes the response addition mixture
model
ON experimental
study
APPENDIX A
Authors
Page 38 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Parent EE;Dence CS;Jenks C;Sharp TL;Welch
MJ;Katzenellenbogen JA;
2007
429
Synthesis and biological evaluation of F-18 bicalutamide,
4- Br-76 bromobicalutamide, and 4- Br-76 bromothiobicalutamide as non-steroidal androgens for prostate
cancer imaging
OFF domain
Park JS;Jun HJ;Cho MJ;Cho KH;Lee JS;Zo
JI;Pyo H;
2006
781
Radiosensitivity enhancement by combined treatment of
celecoxib and gefitinib on human lung cancer cells
OFF domain
Park JY;Elshami AA;Amin K;Rizk N;Kaiser
LR;Albelda SM;
1997
678
Retinoids augment the bystander effect in vitro and in vivo
in herpes simplex virus thymidine kinase ganciclovirmediated gene therapy
OFF domain
Park SW;Kim HS;Hah JH;Kim KH;Heo
DS;Sung MW;
2010
446
DIFFERENTIAL EFFECTS BETWEEN CYCLOOXYGENASE-2
INHIBITORS AND siRNA ON VASCULAR ENDOTHELIAL
GROWTH FACTOR PRODUCTION IN HEAD AND NECK
SQUAMOUS CELL CARCINOMA CELL LINES
OFF domain
Park SW;Kim HS;Hah JW;Jeong WJ;Kim
KH;Sung MW;
2010
652
Celecoxib inhibits cell proliferation through the activation
of ERK and p38 MAPK in head and neck squamous cell
carcinoma cell lines
OFF domain
Parker DC;Wadsworth DC;Schneider GB;
1980
98
ACTIVATION OF MURINE LYMPHOCYTES-B BY ANTIIMMUNOGLOBULIN IS AN INDUCTIVE SIGNAL LEADING
TO IMMUNOGLOBULIN SECRETION
OFF domain
Parrott JL;Sprague JB;
1993
495
PATTERNS IN TOXICITY OF SUBLETHAL MIXTURES OF
METALS AND ORGANIC-CHEMICALS DETERMINED BY
MICROTOX(R) AND BY DNA, RNA, AND PROTEINCONTENT OF FATHEAD MINNOWS (PIMEPHALESPROMELAS)
ON experimental
study
Parvez S;Venkataraman C;Mukherji S;
2009
879
Nature and prevalence of non-additive toxic effects in
industrially relevant mixtures of organic chemicals
ON experimental
study
Patriti A;Facchiano E;Annetti C;Aisa MC;Galli
F;Fanelli C;Donini A;
2005
463
Early improvement of glucose tolerance after ileal
transposition in a non-obese type 2 diabetes rat model
OFF domain
Pavlaki MD;Pereira R;Loureiro S;Soares A;
2011
917
Effects of binary mixtures on the life traits of Daphnia
magna
ON experimental
study
Payne J;Rajapakse N;Wilkins M;Kortenkamp
A;
2000
864
Prediction and assessment of the effects of mixtures of
four xenoestrogens
ON experimental
study
Payne J;Scholze M;Kortenkamp A;
2001
620
Mixtures of four organochlorines enhance human breast
cancer cell proliferation
ON experimental
study
Pearson LJ;Rait C;Nicholls MG;Yandle
TG;Evans JJ;
2006
657
Regulation of adrenomedullin release from human
endothelial cells by sex steroids and angiotensin-II
OFF domain
Pedarzani P;Storm JF;
1995
644
PROTEIN-KINASE A-INDEPENDENT MODULATION OF ION
CHANNELS IN THE BRAIN BY CYCLIC-AMP
OFF domain
Peng T;Zhu J;Hwangbo Y;Corey L;Bumgarner
RE;
2008
1083
Independent and cooperative antiviral actions of beta
interferon and gamma interferon against herpes simplex
virus replication in primary human fibroblasts
OFF domain
Perez-Cruz I;Carcamo JM;Golde DW;
2003
925
Vitamin C inhibits FAS-induced apoptosis in monocytes
and U937 cells
OFF domain
Perloff LJ;Barker CF;
1984
174
VARIABLE RESPONSE TO DONOR-SPECIFIC BLOODTRANSFUSION IN THE RAT
OFF domain
Pesce S;Lissalde S;Lavieille D;Margoum
C;Mazzella N;Roubeix V;Montuelle B;
2010
1049
Evaluation of single and joint toxic effects of diuron and its ON experimental
main metabolites on natural phototrophic biofilms using a
study
pollution-induced community tolerance (PICT) approach
Pestana JLT;Loureiro S;Baird DJ;Soares
AMVM;
2009
946
Fear and loathing in the benthos: Responses of aquatic
insect larvae to the pesticide imidacloprid in the presence
of chemical signals of predation risk
ON experimental
study
Petersen K;Tollefsen KE;
2011
993
Assessing combined toxicity of estrogen receptor agonists
in a primary culture of rainbow trout (Oncorhynchus
mykiss) hepatocytes
ON experimental
study
APPENDIX A
Authors
Page 39 of 66
PubYear
ID
Title
CFT/EFSA/PPR/2010/02
Relevance
Peterson CM;Lu JM;Gu ZW;Shiah JG;Lythgoe
K;Peterson A;Straight RC;Kopecek J;
1995
719
ISOBOLOGRAPHIC ASSESSMENT OF THE INTERACTION
OFF domain
BETWEEN ADRIAMYCIN AND PHOTODYNAMIC THERAPY
WITH MESO-CHLORIN E(6) MONOETHYLENE DIAMINE IN
HUMAN EPITHELIAL OVARIAN-CARCINOMA (OVCAR-3) INVITRO
Petho G;Derow A;Reeh PW;
2001
476
Bradykinin-induced nociceptor sensitization to heat is
mediated by cyclooxygenase products in isolated rat skin
OFF domain
Petrova S;Popov T;
1981
991
Experimental verification of the possibility of in vivo
formation of nitrosocompounds upon combined action of
dodecylguanidineacetate and sodium nitrite
OFF domain
Pezeshki G;Franke B;Engele J;
2001
564
Evidence for a ligand-specific signaling through GFR alpha- OFF domain
1, but not GFR alpha-2, in the absence of ret
Piegorsch WW;Weinberg CR;Haseman JK;
1986
221
TESTING FOR SIMPLE INDEPENDENT ACTION BETWEEN
2 FACTORS FOR DICHOTOMOUS RESPONSE DATA
ON trivial
Piegorsch WW;Weinberg CR;Margolin BH;
1988
269
EXPLORING SIMPLE INDEPENDENT ACTION IN
MULTIFACTOR TABLES OF PROPORTIONS
ON trivial
Pietrowski D;Phan GAB;Tempfer C;Keck C;
2003
720
Description and characterisation of a splicing-variant of
insulin-like growth factor-binding protein-7 (IGFBP-7)
expressed in human granulosa cells
OFF domain
Pignol B;Henane S;Chaumeron S;MenciaHuerta JM;Braquet P;
1990
753
PHARMACOLOGIC MODULATION OF THE EFFECTS OF
PLATELET-ACTIVATING FACTOR ON THE INTERLEUKIN-1
ACTIVITY PRESENT IN SUPERNATANTS FROM
LIPOPOLYSACCHARIDE-STIMULATED RAT SPLEEN
MACROPHAGES
OFF domain
Pipeleers DG;Schuit FC;Vanschravendijk
CFH;Vandewinkel M;
1985
201
INTERPLAY OF NUTRIENTS AND HORMONES IN THE
REGULATION OF GLUCAGON-RELEASE
OFF domain
Pittner RA;Fain JN;
1990
312
VASOPRESSIN AND NOREPINEPHRINE STIMULATION OF
INOSITOL PHOSPHATE ACCUMULATION IN RAT
HEPATOCYTES ARE MODIFIED DIFFERENTLY BY
PROTEIN KINASE-C AND PROTEIN KINASE-A
OFF domain
Poch G;Londong W;
1993
382
MODERN APPROACH TO THE EVALUATION OF
COMBINED EFFECTS OF SINGLE-DOSE TRIALS AND
CLINICAL TIME-COURSE STUDIES, EXEMPLIFIED BY
COMBINATIONS OF PIRENZEPINE AND H-2-RECEPTOR
ANTAGONISTS
ON experimental
study
Podbielski A;Woischnik M;Kreikemeyer
B;Bettenbrock K;Buttaro BA;
1999
570
Cysteine protease SpeB expression in group A streptococci
is influenced by the nutritional environment but SpeB does
not contribute to obtaining essential nutrients
OFF domain
Polc P;
1988
270
ELECTROPHYSIOLOGY OF BENZODIAZEPINE RECEPTOR
LIGANDS - MULTIPLE MECHANISMS AND SITES OF
ACTION
ON trivial
Pons AM;Malo J;Artigas JM;Capilla P;
1999
999
Image quality metric based on multidimensional contrast
perception models
OFF domain
Pons FW;Adlerbollinger A;Muller P;
1993
1123
SYNERGISM AND ANTAGONISM BETWEEN N-METHYL-N'NITRO-N-NITROSO-GUANIDINE AND 9-AMINOACRIDINE
IN MUTATION-INDUCTION IN ESCHERICHIA-COLI K12
OFF domain
Ponzio NM;Speirs RS;
1975
40
LYMPHOID-CELL DEPENDENCE OF EOSINOPHIL
RESPONSE TO ANTIGEN .6. EFFECT OF SELECTIVE
REMOVAL OF T-LYMPHOCYTES OR B-LYMPHOCYTES ON
CAPACITY OF PRIMED SPLEEN-CELLS TO ADOPTIVELY
TRANSFERRED IMMUNITY TO TETANUS TOXOID
OFF domain
Pope RM;McChesney L;Stebbing N;Goldstein
L;Talal N;
1985
202
REGULATION OF T-CELL PROLIFERATION BY CLONED
INTERFERON-ALPHA MEDIATED BY LEU-11B-POSITIVE
CELLS
OFF domain
Popova EP;Lyskovtsev VV;Kaverina NV;
2006
704
Effect of the class III antiarrhythmic drug nibentan on
vagotonic atrial fibrillation in dogs
OFF domain
Popovtzer MM;
1981
119
TUMOR-INDUCED HYPOPHOSPHATEMIC OSTEOMALACIA
(TUO) - EVIDENCE FOR A PHOSPHATURIC CYCLIC AMPINDEPENDENT ACTION OF TUMOR EXTRACT
OFF domain
APPENDIX A
Authors
Page 40 of 66
PubYear
ID
Title
CFT/EFSA/PPR/2010/02
Relevance
Pourshadi M;Defazio SR;Gozzo JJ;
1989
291
ABROGATION OF ALLOREACTIVE SPLEEN CELL-INDUCED OFF domain
2ND-SET SKIN-GRAFT REJECTION IN MICE WITH DONORSPECIFIC BONE-MARROW CELLS
Prabhakar V;Kishore PVK;Balaji M;Balaji
V;Seshaiah V;
2005
447
Can statins be considered as a tertiary level agent in
patients with failure of conventional oral hypoglycemic
agents?
OFF domain
Pravdic D;Mio Y;Sedlic F;Pratt PF;Warltier
DC;Bosnjak ZJ;Bienengraeber M;
2010
439
Isoflurane protects cardiomyocytes and mitochondria by
immediate and cytosol-independent action at reperfusion
OFF domain
Preisler HK;Robertson JL;Hoover
K;McCutchen BF;
1999
425
Statistical methods to assessing responses over time in
bioassays with mixtures
ON method dev.
Preston KR;Tipples KH;
1980
99
EFFECTS OF ACID-SOLUBLE AND ACID-INSOLUBLE
GLUTEN PROTEINS ON THE RHEOLOGICAL AND BAKING
PROPERTIES OF WHEAT FLOURS
OFF domain
Procacci V;Giovine A;Roberto MG;Ronchi
AM;Lasorsa G;Vendemiale G;Stufano
N;Altomare E;
1988
1062
PHARMACODYNAMICS OF SECOND GENERATION
SULFONYLUREAS MECHANISM OF ACTION OF
GLIQUIDONE IN TYPE II DIABETIC PATIENTS
OFF domain
Qin LT;Liu Ss;Zhang J;Xiao QF;
2011
502
A novel model integrated concentration addition with
independent action for the prediction of toxicity of multicomponent mixture
ON method dev.
Qu WD;Zhang TB;Wu DS;Zhang BZ;Deng
Y;Zhou Y;
2000
1066
Combined action of ethanol and acetaldehyde on
embryonic development
OFF domain
R. Koivusalo;E. Krausz;P. Ruotsalainen;H.
Helenius;S. Hietanen
2002
1141
Chemoradiation of cervical cancer cells: Targeting human
papillomavirus E6 and p53 leads to either augmented or
attenuated apoptosis depending on the platinum carrier
ligand
ON trivial
Ra JS;Lee BC;Chang NI;Kim SD;
2006
1024
Estimating the combined toxicity by two-step prediction
model on the complicated chemical mixtures from
wastewater treatment plant effluents
ON method dev.
Raaphorst GP;Azzam EI;Borsa J;Sargent MD;
1985
203
INVITRO TRANSFORMATION BY BROMODEOXYURIDINE
AND X-IRRADIATION IN C3H 10T1/2 CELLS
OFF domain
Rabin RJ;
1974
31
LIMITATIONS ON EMPLOYER INDEPENDENT ACTION
OFF domain
Rabinovich GA;Ramhorst RE;Rubinstein
N;Corigliano A;Daroqui MC;Kier-Joffe
EB;Fainboim L;
2002
558
Induction of allogenic T-cell hyporesponsiveness by
galectin-1-mediated apoptotic and non-apoptotic
mechanisms
OFF domain
Raja A;Ranganathan UD;Bethunaicken R;
2006
1076
Clinical value of specific detection of immune complexbound antibodies in pulmonary tuberculosis
OFF domain
Rajan VN;Nof SY;
1996
505
Cooperation requirements planning (CRP) for
multiprocessors: Optimal assignment and execution
planning
OFF domain
Rajapakse N;Ong D;Kortenkamp A;
2001
1120
Defining the impact of weakly estrogenic chemicals on the
action of steroidal estrogens
ON experimental
study
Rajasekhar VK;Sopory SK;
1985
204
THE BLUE-LIGHT EFFECT AND ITS INTERACTION WITH
PHYTOCHROME IN THE CONTROL OF NITRITE
REDUCTASE-ACTIVITY IN SORGHUM-BICOLOR WILLD
OFF domain
Rak A;Gregoraszczuk EL;
2008
563
Local feedback loop of ghrelin-GH in the pig ovary: Action OFF domain
on estradiol secretion, aromatase activity and cell apoptosis
Ram-Mohan LR;Vurgaftman I;Meyer JR;
1999
1112
Wave function engineering of antimonide quantum-well
lasers
OFF domain
Randall RC;Wilson MA;Setcos JC;Wilson NH;
1998
853
Impression materials and techniques for crown and
bridgework: a survey of undergraduate teaching in the UK
OFF domain
Rapparini F;Rotondi A;Baraldi R;
1999
772
Blue light regulation of the growth of Prunus persica plants OFF domain
in a long term experiment: morphological and histological
observations
Rayburn JR;Friedman M;Bantle JA;
1995
915
Synergistic interaction of glycoalkaloids alpha-chaconine
and alpha-solanine on developmental toxicity in Xenopus
embryos
ON experimental
study
APPENDIX A
Authors
Page 41 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Reantragoon S;Arrigo LM;Dweck
HS;Rosenfeld L;
1996
573
Suppression of endothelin-1 production in cultured human
umbilical vein endothelial cells by heparin fractions
separated by strong anion exchange chromatography
OFF domain
Rebecchi MJ;Gershengorn MC;
1983
158
THYROLIBERIN STIMULATES RAPID HYDROLYSIS OF
PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE BY A
PHOSPHODIESTERASE IN RAT MAMMOTROPIC
PITUITARY-CELLS - EVIDENCE FOR AN EARLY CA-2+INDEPENDENT ACTION
OFF domain
Reed JW;Nagpal P;Bastow RM;Solomon
KS;Dowson-Day MJ;Elumalai RP;Millar AJ;
2000
664
Independent action of ELF3 and phyB to control hypocotyl
elongation and flowering time
OFF domain
Rice G;Teuschler LK;Speth TF;Richardson
SD;Miltner RJ;Schenck KM;Gennings
C;Hunter ES;Narotsky MG;Simmons JE;
2008
1042
Integrated disinfection by-products research: Assessing
reproductive and developmental risks posed by complex
disinfection by-product mixtures
ON experimental
study
Richardson WK;Clark DB;
1976
51
COMPARISON OF KEY-PECK AND TREADLE-PRESS
OPERANTS IN PIGEON - DIFFERENTIALREINFORCEMENT-OF-LOW-RATE SCHEDULE OF
REINFORCEMENT
OFF domain
Richter M;Escher BI;
2005
977
Mixture toxicity of reactive chemicals by using two
bacterial growth assays as indicators of protein and DNA
damage
ON experimental
study
Rider CV;LeBlanc GA;
2005
934
An integrated addition and interaction model for assessing
toxicity of chemical mixtures
ON method dev.
Rider CV;LeBlanc GA;
2006
572
Atrazine stimulates hemoglobin accumulation in Daphnia
magna: Is it hormonal or hypoxic?
OFF domain
Riedl E;Strobl H;Majdic O;Knapp W;
1997
739
TGF-beta 1 promotes in vitro generation of dendritic cells
by protecting progenitor cells from apoptosis
OFF domain
Riss TL;Karey KP;Burleigh BD;Parker
D;Sirbasku DA;
1988
271
HUMAN RECOMBINANT INSULIN-LIKE GROWTH FACTORI .1. DEVELOPMENT OF A SERUM-FREE MEDIUM FOR
CLONAL DENSITY ASSAY OF GROWTH-FACTORS USING
BALB/C 3T3 MOUSE EMBRYO FIBROBLASTS
OFF domain
Robertson JL;Smith KC;
1984
176
WESTERN SPRUCE BUDWORM - JOINT ACTION OF
PYRETHROIDS AND INSECT GROWTH-REGULATORS BY
CONTACT OR INGESTION
ON trivial
Robertson JL;Smith KC;
1984
175
JOINT ACTION OF PYRETHROIDS WITH ORGANOPHOSPHORUS AND CARBAMATE INSECTICIDES APPLIED
TO WESTERN SPRUCE BUDWORM (LEPIDOPTERA,
TORTRICIDAE)
ON trivial
Robertson JL;Smith KC;Granett J;Retnakaran
A;
1984
177
JOINT ACTION OF A JUVENILE-HORMONE ANALOG WITH
BENZOYLPHENYLUREAS INGESTED BY WESTERN SPRUCE
BUDWORM, CHORISTONEURA-OCCIDENTALIS
(LEPIDOPTERA, TORTRICIDAE)
ON trivial
Roche JP;Westenbroek R;Sorom AJ;Hille
B;Mackie K;Shapiro MS;
2002
350
Antibodies and a cysteine-modifying reagent show
correspondence of M current in neurons to KCNQ2 and
KCNQ3 K+ channels
OFF domain
Rochetaing A;Barbe C;Kreher P;
2001
909
Beneficial effects of amiodarone and dronedarone (SR
33589b), when applied during low-flow ischemia, on
arrhythmia and functional parameters assessed during
reperfusion in isolated rat hearts
OFF domain
Rodriguez JE;Moninger T;Grose C;
1993
326
ENTRY AND EGRESS OF VARICELLA VIRUS BLOCKED BY
SAME ANTI-GH MONOCLONAL-ANTIBODY
OFF domain
Rogulj N;Srhoj V;Srhoj L;
2004
997
The contribution of collective attack tactics in
differentiating handball score efficiency
OFF domain
Rolapleszczynski M;Gagnon L;Bolduc
D;Lebreton G;
1985
205
EVIDENCE FOR THE INVOLVEMENT OF THE
THROMBOXANE SYNTHASE PATHWAY IN HUMAN
NATURAL CYTO-TOXIC CELL-ACTIVITY
OFF domain
Rorsman P;Hellman B;
1983
159
THE INTERACTION BETWEEN MANGANESE AND CALCIUM OFF domain
FLUXES IN PANCREATIC BETA-CELLS
APPENDIX A
Authors
Page 42 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Rosengren A;Filipsson K;Ahren B;Renstrom E;
2001
335
Pituitary adenylate cyclase-activating polypeptide mimicks
the ATP-sensitve K-channel-independent action of glucose
at low glucose concentrations
OFF domain
Roswall P;Bu SZ;Rubin K;Landstrom M;Heldin
NE;
2006
427
2-methoxyestradiol induces apoptosis in cultured human
anaplastic thyroid carcinoma cells
OFF domain
Rottiers W;
2008
615
Sound absorption with fibre-free sintered aluminium in
combination with thermally activated concrete slabs
OFF domain
Roy P;Chatterjee A;Banerjee PP;Bhattacharya
S;
2000
695
A thyrotropin-like molecule from the pituitary of an Indian
freshwater murrel: comparison of its biological activity
with other thyrotropins
OFF domain
Roy RC;Stulken EH;
1981
120
ELECTROENCEPHALOGRAPHIC EVIDENCE OF AROUSAL
IN DOGS FROM HALOTHANE AFTER DOXAPRAM,
PHYSOSTIGMINE, OR NALOXONE
OFF domain
Royle JS;Ross J;Ansell IF;Bollina P;Habib F;
2003
339
Nitric oxide non-steroidal anti-inflammatory drugs induce
apoptosis by CGMP independent means in prostate cell
systems
OFF domain
Rudzok S;Schlink U;Herbarth O;Bauer M;
2010
956
Measuring and modeling of binary mixture effects of
pharmaceuticals and nickel on cell viability/cytotoxicity in
the human hepatoma derived cell line HepG2
ON experimental
study
Ryu BS;Yang HS;
1999
384
Integration of reactive behaviors and enhanced topological OFF domain
map for robust mobile robot navigation
S. Chaperon;S. Sauve
2007
1133
Toxicity interaction of metals (Ag, Cu, Hg, Zn) to urease
and dehydrogenase activities in soils
ON trivial
S. Chaperon;S. Sauve
2008
1134
Toxicity interactions of cadmium, copper, and lead on soil
urease and dehydrogenase activity in relation to chemical
speciation
ON experimental
study
Sachs G;Wallmark B;
1989
1035
Biological basis of omeprazole therapy
OFF domain
Sagastagoitia JD;Morillas M;Martinez A;Lage
E;Pallares C;Ugartemendia C;Asin E;Barrios
V;Julia J;Sanchez P;Calvo C;Gonzalez J;Pina
C;Garcia-Barbal J;Arnau C;
1998
423
Improvement in diastolic function in hypertensive patients
with left ventricular hypertrophy with inhibitors of the
angiotensin converting enzyme
OFF domain
Saito Y;Nishio K;Ogawa Y;Kinumi T;Yoshida
Y;Masuo Y;Niki E;
2007
983
Molecular mechanisms of 6-hydroxydopamine-induced
cytotoxicity in PC12 cells: Involvement of hydrogen
peroxide-dependent and -independent action
OFF domain
Sakamoto K;Elkind MM;
1969
12
X-RAYS AND NITROGEN MUSTARD - INDEPENDENT
ACTION IN CHINESE HAMSTER CELLS
OFF domain
Salam S;Cox JF;Sims HP;
1997
1044
In the eye of the beholder - How leadership relates to 360- OFF domain
degree performance ratings
Salama HS;Foda MS;Sharaby A;
1983
160
BIOLOGICAL-ACTIVITY OF MIXTURES OF BACILLUSTHURINGIENSIS VARIETIES AGAINST SOME COTTON
PESTS
OFF domain
Samaha RR;Boyle TH;
1989
292
SELF-INCOMPATIBILITY OF ZINNIA-ANGUSTIFOLIA HBK
(COMPOSITAE) .2. GENETICS
OFF domain
Samokhin GP;Minin AA;Bespalova JD;Titov
MI;Smirnov VN;
1981
121
INDEPENDENT ACTION OF ALPHA-FACTOR AND
CYCLOHEXIMIDE ON THE RATE OF CELL-CYCLE
INITIATION IN SACCHAROMYCES-CEREVISIAE
OFF domain
Sanchez JA;Dani JA;Siemen D;Hille B;
1986
222
SLOW PERMEATION OF ORGANIC CATIONS IN
ACETYLCHOLINE-RECEPTOR CHANNELS
OFF domain
Sanchez M;Picard N;Sauve K;Tremblay A;
2010
554
Challenging estrogen receptor beta with phosphorylation
OFF domain
Santiano N;Young L;Hillman K;Parr
M;Jayasinghe S;Baramy LS;Stevenson
J;Heath T;Chan C;Claire M;Hanger G;
2009
724
Analysis of Medical Emergency Team calls comparing
subjective to 'objective' call criteria
OFF domain
Santos MJG;Soares A;Loureiro S;
2010
910
Joint effects of three plant protection products to the
terrestrial isopod Porcellionides pruinosus and the
collembolan Folsomia candida
ON experimental
study
Sapers GM;Panasiuk O;Carre J;
1978
70
EFFECTS OF THERMAL PROCESSING AND SALT ON PH
AND ACIDITY OF HOME CANNED TOMATOES
OFF domain
APPENDIX A
Authors
Page 43 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
OFF domain
Saperstein AM;
1992
419
ALLIANCE BUILDING VERSUS INDEPENDENT ACTION - A
NONLINEAR MODELING APPROACH TO COMPARATIVE
INTERNATIONAL STABILITY
Sato E;Mokudai T;Niwano Y;Kamibayashi
M;Kohno M;
2008
1111
Existence of a new reactive intermediate oxygen species in OFF domain
hypoxanthine and xanthine oxidase reaction
Sato Y;Henquin JC;
1998
480
The K+-ATP channel-independent pathway of regulation
of insulin secretion by glucose - In search of the
underlying mechanism
OFF domain
Savard K;Berthelot Y;Auroy A;Spear
PA;Trottier B;Robidoux PY;
2007
576
Effects of HMX - Lead mixtures on reproduction of the
earthworm Eisenia andrei
OFF domain
Sawhney RN;
1981
122
SOME COMMENTS ON 2 RECENT PAPERS IN EUPHYTICA
CONCERNING STEM RUST RESISTANCE IN WHEAT RESPONSE
OFF domain
Scatena R;Bottoni P;Giardina B;
2008
467
Mitochondria, PPARs, and Cancer: Is ReceptorIndependent Action of PPAR Agonists a Key?
ON review
Schar J;Sickmann A;Beier D;
2005
931
Phosphorylation-independent activity of atypical response
regulators of Helicobacter pylori
OFF domain
Schenkelaars EJ;Bonta IL;
1984
178
BETA-2-ADRENOCEPTOR AGONISTS REVERSE THE
LEUKOTRIENE-C4-INDUCED RELEASE RESPONSE OF
MACROPHAGES
OFF domain
Schlyter F;Birgersson G;Byers JA;Lofqvist
J;Bergstrom G;
1987
241
FIELD RESPONSE OF SPRUCE BARK BEETLE, IPSTYPOGRAPHUS, (COLEOPTERA, SCOLYTIDAE) TO
AGGREGATION PHEROMONE CANDIDATES
OFF domain
Schmeisch AP;Pagadigorria CS;Constantin
J;Vicentini G;Ishii-Iwamoto EL;Bracht A;
1998
933
The role of hemodynamics in the action of diltiazem on
hepatic fatty acid metabolism
OFF domain
Schnabel P;Mies F;Nohr T;Geisler M;Bohm M;
2000
485
Differential regulation of phospholipase C-beta isozymes in
cardiomyocyte hypertrophy
OFF domain
Schochett P;Mora R;Mark L;Butler M;Ingenito
EP;
1999
904
Calcium-dependent degradation of surfactant protein A by
activated neutrophils due to serine proteases
OFF domain
Schroder HC;Trolltsch D;Friese U;Bachmann
M;Muller WEG;
1987
242
MATURE MESSENGER-RNA IS SELECTIVELY RELEASED
FROM THE NUCLEAR MATRIX BY AN ATP/DATPDEPENDENT MECHANISM SENSITIVE TO
TOPOISOMERASE INHIBITORS
OFF domain
Schultz JR;Ellerby LM;Gralla EB;Valentine
JS;Clarke CF;
1996
1085
Autoxidation of ubiquinol-6 is independent of superoxide
dismutase
OFF domain
Schurch D;Ospina OL;Cruz A;Perez-Gil J;
2010
938
Combined and Independent Action of Proteins SP-B and
SP-C in the Surface Behavior and Mechanical Stability of
Pulmonary Surfactant Films
OFF domain
Schurov IL;Hepworth TJ;Hastings MH;
2002
872
Dopaminergic signalling in the rodent neonatal
suprachiasmatic nucleus identifies a role for protein kinase
A and mitogen-activated protein kinase in circadian
entrainment
OFF domain
Schuster A;Fahy JV;Ueki I;Nadel JA;
1995
575
CYSTIC-FIBROSIS SPUTUM INDUCES A SECRETORY
RESPONSE FROM AIRWAY GLAND SEROUS CELLS THAT
CAN BE PREVENTED BY NEUTROPHIL PROTEASE
INHIBITORS
OFF domain
Schwartz EL;Chamberlin H;Brechbuhl AB;
1990
313
INHIBITION OF C-MYC EXPRESSION IN HUMAN
PROMYELOCYTIC LEUKEMIA AND COLON
ADENOCARCINOMA CELLS BY 6-THIOGUANINE
OFF domain
Scibior A;Zaporowska H;Niedzwiecka I;
2009
903
Lipid peroxidation in the liver of rats treated with V and/or
Mg in drinking water
ON experimental
study
Scornik FS;Codina J;Birnbaumer L;Toro L;
1993
327
INDEPENDENT ACTION OF G-PROTEIN AND PKADEPENDENT PHOSPHORYLATION UNDERLIE BETAADRENOCEPTOR ACTIVATION OF KCA CHANNELS
ON trivial
Scott JA;Hodson PV;
2008
402
Evidence for multiple mechanisms of toxicity in larval
rainbow trout (Oncorhynchus mykiss) co-treated with
retene and alpha-naphthoflavone
ON experimental
study
APPENDIX A
Authors
Page 44 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Sekharam M;Cunnick JM;Wu J;
2000
408
Involvement of lipoxygenase in lysophosphatidic acidstimulated hydrogen peroxide release in human HaCaT
keratinocytes
OFF domain
Sepulveda N;Paulino CD;Carneiro J;PenhaGoncalves C;
2007
673
Allelic penetrance approach as a tool to model two-locus
interaction in complex binary traits
OFF domain
Serlin BS;Sopory SK;Roux SJ;
1984
179
MODULATION OF OAT MITOCHONDRIAL ATPASE
ACTIVITY BY CA-2+ AND PHYTOCHROME
OFF domain
Severina IS;
1980
952
MECHANISMS OF INHIBITION BY CHLORGYLINE AND
DEPRENYL OF TYRAMINE OXIDATIVE DEAMINATION BY
RAT LIVER MITOCHONDRIAL MONO AMINE OXIDASE
OFF domain
Shadbolt CT;Ross T;McMeekin TA;
1999
885
Nonthermal death of Escherichia coli
OFF domain
Shankar G;Stern PH;
1993
1073
EVALUATION OF THE ROLE OF 2ND MESSENGER
SYSTEMS IN TUMOR NECROSIS FACTOR-STIMULATED
RESORPTION OF FETAL-RAT LIMB BONES
OFF domain
Sharaf AA;el-Sherbini A;Abdulla WA;
1976
469
Metabolic effects of some antibilharzial drugs inhibition of
citrate oxidation by Bilharcid
OFF domain
Sharma SS;Schat H;Vooijs R;Van
Heerwaarden LM;
1999
1090
Combination toxicology of copper, zinc, and cadmium in
binary mixtures: Concentration-dependent antagonistic,
nonadditive, and synergistic effects on root growth in
Silene vulgaris
ON experimental
study
Shaw MW;
1989
293
INDEPENDENT ACTION OF FUNGICIDES AND ITS
CONSEQUENCES FOR STRATEGIES TO RETARD THE
EVOLUTION OF FUNGICIDE RESISTANCE
ON trivial
Shaw MW;
1993
371
THEORETICAL-ANALYSIS OF THE EFFECT OF
INTERACTING ACTIVITIES ON THE RATE OF SELECTION
FOR COMBINED RESISTANCE TO FUNGICIDE MIXTURES
ON trivial
Shcherbakov VP;Kudryashova
EA;Shcherbakova TS;Sizova ST;Plugina LA;
2006
1007
Double-strand break repair in bacteriophage T4:
Recombination effects of 3 '-5 ' exonuclease mutations
OFF domain
Shen Z;Saloniemi T;Ronnblad A;Jarvensivu
P;Pakarinen P;Poutanen M;
2009
1108
Sex Steroid-Dependent and -Independent Action of
Hydroxysteroid (17 beta) Dehydrogenase 2: Evidence
from Transgenic Female Mice
OFF domain
Shetty AK;Turner DA;
1998
701
In vitro survival and differentiation of neurons derived
from epidermal growth factor-responsive postnatal
hippocampal stem cells: Inducing effects of brain-derived
neurotrophic factor
OFF domain
Shiau CW;Yang CC;Kulp SK;Chen KF;Chen
CS;Huang JW;
2005
490
Thiazolidenediones mediate apoptosis in prostate cancer
cells in part through inhibition of Bcl-xL/Bcl-2 functions
independently of PPAR-gamma
OFF domain
Shibuya H;Sakai K;Kabir-Salmani M;Wachi
Y;Iwashita M;
2011
634
Polymerization of Insulin-Like Growth Factor-Binding
Protein-1 (IGFBP-1) Potentiates IGF-I Actions in Placenta
OFF domain
Shields SK;Nicola C;Chakraborty C;
2007
474
Rho guanosine 5 '-triphosphatases differentially regulate
OFF domain
insulin-like growth factor I (IGF-I) receptor-ependent and independent actions of IGF-II on human trophoblast
migration
Shinozuka H;Lombardi B;Abanobi SE;
1982
143
A COMPARATIVE-STUDY OF THE EFFICACY OF 4
BARBITURATES AS PROMOTERS OF THE DEVELOPMENT
OF GAMMA-GLUTAMYLTRANSPEPTIDASE-POSITIVE FOCI
IN THE LIVER OF CARCINOGEN TREATED RATS
OFF domain
Shirit Einav;Hadas Dvory-Sobol;Elizabeth
Gehrig;Jeffrey S. Glenn
2010
1138
The Hepatitis C Virus (HCV) NS4B RNA Binding Inhibitor
Clemizole Is Highly Synergistic with HCV Protease
Inhibitors
ON experimental
study
Shirwaikar A;Rajendran K;Punitha ISR;
2005
861
Antihyperglycemic activity of the aqueous stem extract of
Coscinium fenestratum in non-insulin dependent diabetic
rats
OFF domain
Shojikasai Y;Yoshida A;Ogura A;Kuwahara
R;Grasso A;Takahashi M;
1994
405
SYNAPTOTAGMIN-I IS ESSENTIAL FOR CA2+INDEPENDENT RELEASE OF NEUROTRANSMITTER
INDUCED BY ALPHA-LATROTOXIN
OFF domain
APPENDIX A
Authors
Page 45 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Shtabsky BM;Fedorenko VI;Kandul SV;Kagan
YS;
1995
874
COMBINATORY TOXICITY OF SOME AGROCHEMICAL AND
HEAVY-METALS FOR RATS COMMUNICATION .2.
TRICHLORPHON, LEAD, NITRATES, LINDANE AND
CADMIUM
ON experimental
study
Sidell N;Verity MA;Nord EP;
1990
314
MENTHOL BLOCKS DIHYDROPYRIDINE-INSENSITIVE CA2+ CHANNELS AND INDUCES NEURITE OUTGROWTH IN
HUMAN NEUROBLASTOMA-CELLS
OFF domain
Sidobre S;Naidenko OV;Sim BC;Gascoigne
NRJ;Garcia KC;Kronenberg M;
2002
691
The V alpha 14 NKT cell TCR exhibits high-affinity binding
to a glycolipid/CD1d complex
OFF domain
Sigurd B;Olesen KH;
1977
59
SUPRA-ADDITIVE NATRIURETIC EFFECT ADDITION OF
THEOPHYLLINE ETHYLENEDIAMINE AND BUMETANIDE
IN CONGESTIVE HEART-FAILURE - PERMUTATION TRIAL
TESTS IN PATIENTS IN LONG-TERM TREATMENT WITH
BUMETANIDE
OFF domain
Sigurd B;Olesen KH;Wennevold A;
1975
41
SUPRA-ADDITIVE NATRIURETIC EFFECT ADDITION OF
BENDROFLUMETHIAZIDE AND BUMETANIDE IN
CONGESTIVE HEART-FAILURE - PERMUTATION TRIAL
TESTS IN PATIENTS IN LONG-TERM TREATMENT WITH
BUMETANIDE
OFF domain
Sikka R;Kansal BD;
1995
379
EFFECT OF FLY-ASH APPLICATION ON YIELD AND
NUTRIENT COMPOSITION OF RICE, WHEAT AND ON PH
AND AVAILABLE NUTRIENT STATUS OF SOILS
OFF domain
Silva E;Rajapakse N;Kortenkamp A;
2002
1121
Something from 'nothing' - Eight weak estrogenic
chemicals combined at concentrations below NOECs
produce significant mixture effects
ON experimental
study
Simpson LG;Isakson PC;
1986
223
ROLE OF DNA-SYNTHESIS IN SECRETION OF
IMMUNOGLOBULIN FROM MURINE B-CELLS STIMULATED
BY T-CELL DERIVED LYMPHOKINES
OFF domain
Sitges M;Nekrassov V;Guarneros A;
2000
1015
Simultaneous action of MK-801 (dizclopine) on dopamine,
glutamate, aspartate and GABA release from striatum
isolated nerve endings
OFF domain
Sjogren C;Andersson KE;Husted S;Mattiasson
A;Mollermadsen B;
1982
144
ATROPINE RESISTANCE OF TRANSMURALLY
STIMULATED ISOLATED HUMAN BLADDER MUSCLE
OFF domain
Skorija K;Cox M;Sisk JM;Dowd DR;MacDonald
PN;Thompson CC;Demay MB;
2005
404
Ligand-independent actions of the vitamin D receptor
maintain hair follicle homeostasis
OFF domain
Skott O;
1987
243
EFFECTS OF AMINES, MONENSIN AND NIGERICIN ON
THE RENIN RELEASE FROM ISOLATED SUPERFUSED RAT
GLOMERULI
OFF domain
Smit MJ;Roovers E;Timmerman
H;vandeVrede Y;Alewijnse AE;Leurs R;
1996
763
Two distinct pathways for histamine H-2 receptor downregulation
OFF domain
Smith CIE;Hammarstrom L;
1978
71
SODIUM POLYANETHOLE SULFONATE - NEW
MACROPHAGE-DEPENDENT POLYMYXIN-INHIBITABLE,
POLYCLONAL B-CELL ACTIVATOR
OFF domain
Smith D;Breeden L;Farrell E;Yarus M;
1987
244
THE BASES OF THE TRANSFER-RNA ANTICODON LOOP
ARE INDEPENDENT BY GENETIC CRITERIA
OFF domain
Smith DO;Lu Z;
1991
651
ADENOSINE DERIVED FROM HYDROLYSIS OF
PRESYNAPTICALLY RELEASED ATP INHIBITS
NEUROMUSCULAR-TRANSMISSION IN THE RAT
OFF domain
Snippe H;Kamp E;
1975
42
CELL COOPERATION AND HAPTEN-CARRIER COMPLEXES
OFF domain
Sobye KW;Streibig JC;Cedergreen N;
2011
742
Prediction of joint herbicide action by biomass and
chlorophyll a fluorescence
ON experimental
study
Solana RP;Carter WH;Wilson JD;Chinchilli
VM;Carchman RA;
1988
272
QUALITATIVE EVALUATION OF SISTER CHROMATID
EXCHANGES ELICITED BY COMBINATIONS OF
GENOTOXIC COMPOUNDS USING RESPONSE-SURFACE
METHODOLOGY
OFF domain
Solomon PR;Lohr AC;Moore JW;
1974
32
LATENT INHIBITION OF RABBITS NICTITATINGMEMBRANE RESPONSE - SUMMATION TESTS FOR
ACTIVE INHIBITION AS A FUNCTION OF NUMBER OF CS
PRE-EXPOSURES
OFF domain
APPENDIX A
Authors
Page 46 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Somers KD;
1980
100
INCREASED CYCLIC-AMP CONTENT DIRECTLY
CORRELATED WITH MORPHOLOGICAL
TRANSFORMATION OF CELLS INFECTED WITH A
TEMPERATURE-SENSITIVE MUTANT OF MOUSE
SARCOMA-VIRUS
OFF domain
Sondossi M;Riha VF;Rossmoore HW;Sylvestre
M;
1993
533
FACTORS INVOLVED IN BACTERICIDAL ACTIVITIES OF
FORMALDEHYDE AND FORMALDEHYDE CONDENSATE
ISOTHIAZOLONE MIXTURES
OFF domain
Song JY;Kim KD;Roe JH;
2008
1041
Thiol-Independent Action of Mitochondrial Thioredoxin To
Support the Urea Cycle of Arginine Biosynthesis in
Schizosaccharomyces pombe
OFF domain
Song Xq;Liu Ss;Liu Hl;Ge Hl;
2008
684
Mixture toxicity of herbicides and heavy metal compounds
to photobacteria (Vibrio qinghaiensis sp.-Q67)
ON experimental
study
Sopori M;Bernstein A;Bach FH;
1978
72
INVITRO SENSITIZATION OF THYMOCYTES - ROLE OF H2 I REGION DETERMINANTS AND CELL-FREE MIXED
LEUKOCYTE-CULTURE SUPERNATES IN GENERATION OF
CYTOTOXIC RESPONSES
OFF domain
Spagnoli A;Torello M;Nagalla SR;Horton
WA;Pattee P;Hwa V;Chiarelli F;Roberts
CT;Rosenfeld RG;
2002
871
Identification of STAT-1 as a molecular target of IGFBP-3
in the process of chondrogenesis
OFF domain
Sparfel L;Van Grevenynghe J;Le Vee
M;Aninat C;Fardel O;
2006
751
Potent inhibition of carcinogen-bioactivating cytochrome
P4501B1 by the p53 inhibitor pifithrin alpha
OFF domain
Sperling W;Schimz A;
1980
101
PHOTOSENSORY RETINAL PIGMENTS IN
HALOBACTERIUM-HALOBIUM
OFF domain
Spliid H;Torslov J;
1994
866
STATISTICAL-ANALYSIS OF JOINT TOXICITY IN
BIOLOGICAL GROWTH EXPERIMENTS
ON experimental
study
Spurgeon DJ;Jones OAH;Dorne J;Svendsen
C;Swain S;Sturzenbaum SR;
2010
536
Systems toxicology approaches for understanding the joint
effects of environmental chemical mixtures
ON experimental
study
Sritharan S;Priestley MJN;Seible F;
2000
547
Nonlinear finite element analyses of concrete bridge joint
systems subjected to seismic actions
OFF domain
Stanley D;McGrath BJ;Lamp KC;Rybak MJ;
1994
717
EFFECT OF HUMAN SERUM ON KILLING ACTIVITY OF
VANCOMYCIN AND TEICOPLANIN AGAINST
STAPHYLOCOCCUS-AUREUS
OFF domain
Steen VM;Holmsen H;
1985
206
SYNERGISM BETWEEN THROMBIN AND EPINEPHRINE IN
HUMAN-PLATELETS - DIFFERENT DOSE-RESPONSE
RELATIONSHIPS FOR AGGREGATION AND DENSE
GRANULE SECRETION
ON trivial
Stemkowski PL;Tse FW;Peuckmann V;Ford
CP;Colmers WF;Smith PA;
2002
829
ATP-inhibition of M current in frog sympathetic neurons
involves phospholipase C but not Ins P-3, Ca2+, PKC, or
Ras
OFF domain
Stephen FD;Yokota SJ;Repasky EA;
1990
765
THE EFFECT OF FREE FATTY-ACIDS ON SPECTRIN
ORGANIZATION IN LYMPHOCYTES
OFF domain
Stevens J;Stevens YW;
1975
43
SEQUENTIAL IRREVERSIBLE, ACTINOMYCIN DSENSITIVE, AND CYCLOHEXIMIDE-SENSITIVE STEPS
PRIOR TO CORTISOL INHIBITION OF URIDINE
UTILIZATION BY P1798 TUMOR LYMPHOCYTES
OFF domain
Stevenson RW;Stebbing N;Jones T;Carr
K;Jones PM;Hii C;Cherrington AD;
1988
273
THE SYNTHETIC HUMAN GROWTH-HORMONE
FRAGMENT (32-38) INCREASES GLUCOSE-UPTAKE IN
THE CONSCIOUS DOG
OFF domain
Stiffel C;Liacopoulosbriot M;Decreusefond
C;Parlebas J;
1987
245
GENETIC DIFFERENCE IN THE PROLIFERATIVE
RESPONSE TO T-MITOGENS BETWEEN HI/PHA AND
LO/PHA LYMPHOCYTES IS INDEPENDENT OF ACCESSORY
CELL-FUNCTION
OFF domain
Stoddard PK;Markham MR;
2008
527
Signal cloaking by electric fish
OFF domain
Stone ND;Makela ME;Plapp FW;
1988
274
NONLINEAR OPTIMIZATION ANALYSIS OF INSECTICIDE
MIXTURES FOR THE CONTROL OF THE TOBACCO
BUDWORM (LEPIDOPTERA, NOCTUIDAE)
OFF domain
APPENDIX A
Authors
Page 47 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Stover BJ;Eyring H;
1970
13
DYNAMICS OF LIFE, .3. MECHANISMS OF NONSURVIVAL
AND RELATION OF DOSE SIZE
OFF domain
Strausbaugh CA;Myers JR;Forster RL;McClean
PE;
2003
390
A quantitative method to screen common bean plants for
resistance to Bean common mosaic necrosis virus
OFF domain
Stronegger WJ;Berghold A;Seeber GUH;
1998
515
Epidemiological and statistical models of interaction and
their analysis by regression models
OFF domain
Sugaya M;Nakamura K;Asahina A;Fujita
H;Tada Y;Torii H;Tamaki K;
2005
1115
Signaling through CD40 ligand decreases CD80 expression
on murine Langerhans cells and enhances IL-12 p40
production
OFF domain
Sugiura K;Goto M;
1981
123
MUTAGENICITIES OF STYRENE OXIDE DERIVATIVES ON
BACTERIAL TEST SYSTEMS - RELATIONSHIP BETWEEN
MUTAGENIC POTENCIES AND CHEMICAL-REACTIVITY
OFF domain
Sugiyama M;Imai A;Furui T;Tamaya T;
2003
806
Independent action of serine/threonine protein
OFF domain
phosphatase in ovarian cancer plasma membrane and
cytosol during gonadotropin-releasing hormone stimulation
Suh DY;Kagami J;Fukuma K;Iwanami
N;Yamazaki Y;Yurimoto H;Sakai Y;Kato
N;Shibuya M;Ebizuka Y;Sankawa U;
2000
491
Chalcone and stilbene synthases expressed in eucaryotes
exhibit reduced cross-reactivity in vitro
OFF domain
Sullivan GW;Linden J;Hewlett EL;Carper
HT;Hylton JB;Mandell GL;
1990
315
ADENOSINE AND RELATED-COMPOUNDS COUNTERACT
TUMOR NECROSIS FACTOR-ALPHA INHIBITION OF
NEUTROPHIL MIGRATION - IMPLICATION OF A NOVEL
CYCLIC AMP-INDEPENDENT ACTION ON THE CELLSURFACE
OFF domain
Suri S;Liu XH;Rayment S;Hughes DA;Kroon
PA;Needs PW;Taylor MA;Tribolo S;Wilson VG;
2010
440
Quercetin and its major metabolites selectively modulate
OFF domain
cyclic GMP-dependent relaxations and associated tolerance
in pig isolated coronary artery
Surin AM;Bolshakov AP;Mikhailova
MM;Sorokina EG;Senilova YE;Pinelis
VG;Khodorov BI;
2006
675
Arachidonic acid enhances intracellular Ca2+ (i) increase
and mitochondrial depolarization induced by glutamate in
cerebellar granule cells
OFF domain
Sutton CA;Martin TFJ;
1982
145
THYROTROPIN-RELEASING-HORMONE (TRH)
SELECTIVELY AND RAPIDLY STIMULATES
PHOSPHATIDYLINOSITOL TURNOVER IN GH PITUITARYCELLS - A POSSIBLE 2ND STEP OF TRH ACTION
OFF domain
Svendsen C;Siang P;Lister LJ;Rice
A;Spurgeon DJ;
2010
635
SIMILARITY, INDEPENDENCE, OR INTERACTION FOR
BINARY MIXTURE EFFECTS OF NERVE TOXICANTS FOR
THE NEMATODE CAENORHABDITIS ELEGANS
ON experimental
study
Swope SL;Schonbrunn A;
1988
275
THE BIPHASIC STIMULATION OF INSULIN-SECRETION BY OFF domain
BOMBESIN INVOLVES BOTH CYTOSOLIC FREE CALCIUM
AND PROTEIN KINASE-C
Syberg K;Elleby A;Pedersen H;Cedergreen
N;Forbes VE;
2008
685
Mixture toxicity of three toxicants with similar and
dissimilar modes of action to Daphnia magna
ON experimental
study
Szabo M;Staib NE;Collins BJ;Cuttler L;
1990
316
BIPHASIC ACTION OF FORSKOLIN ON GROWTHHORMONE AND PROLACTIN SECRETION BY RAT
ANTERIOR-PITUITARY-CELLS INVITRO
OFF domain
Tachon P;Giacomoni PU;Brandi G;Cantoni O;
1994
936
DIFFERENTIAL-EFFECTS OF HISTIDINE ON HYDROGEN
PEROXIDE-INDUCED BACTERIAL KILLING AND DNA
NICKING IN-VITRO
OFF domain
Tahmatzopoulos A;Kyprianou N;
2004
445
Apoptotic impact of alpha-blockers on prostate cancer
growth: A myth or an inviting reality?
OFF domain
Takagi S;Inada M;Okawa H;Yata J;
1988
943
DIFFERENTIAL EFFECTS OF DIBUTYRYL CYCLIC AMP AND OFF domain
SIMPLE SUGARS ON NK AND LAK ACTIVITIES
SUGGESTING DIFFERENCES OF THEIR CYTOTOXIC
MECHANISM
Takahashi K;Katakura R;Mashiyama
S;Yoshimoto T;Suzuki J;Sasaki T;
1991
787
EFFECT OF X-RAYS COMBINED WITH ACNU AND O-6
ETHYLGUANINE ON RAT SUBCUTANEOUS GLIOMAS
OFF domain
Takahashi KI;Copenhagen DR;
1992
360
APB SUPPRESSES SYNAPTIC INPUT TO RETINAL
HORIZONTAL CELLS IN FISH - A DIRECT ACTION ON
HORIZONTAL CELLS MODULATED BY INTRACELLULAR-PH
OFF domain
APPENDIX A
Authors
Page 48 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Takahashi M;Przetakiewicz M;Ong A;Borek
C;Lowenstein JM;
1992
693
EFFECT OF OMEGA-3 AND OMEGA-6 FATTY-ACIDS ON
TRANSFORMATION OF CULTURED-CELLS BY
IRRADIATION AND TRANSFECTION
OFF domain
Takahashi MA;Gross EL;
1978
73
USE OF IMMOBILIZED LIGHT-HARVESTING
CHLOROPHYLL A-B PROTEIN TO STUDY STOICHIOMETRY
OF ITS SELF-ASSOCIATION
OFF domain
Takenouchi T;Munekata E;
1998
1117
Serotonin increases cytoplasmic Ca2+ concentration in
PC12h cells: effect of tachykinin peptides
OFF domain
Takuma K;Fujita T;Kimura Y;Tanabe
M;Yamamuro A;Lee E;Mori K;Koyama Y;Baba
A;Matsuda T;
2000
893
T-588 inhibits astrocyte apoptosis via mitogen-activated
protein kinase signal pathway
OFF domain
Tallgren LG;
1980
391
INORGANIC SULFATES IN RELATION TO THE SERUM
THYROXINE LEVEL AND IN RENAL FAILURE
OFF domain
Tam VH;Schilling AN;Lewis RE;Melnick
DA;Boucher AN;
2004
1037
Novel approach to characterization of combined
pharmacodynamic effects of antimicrobial agents
ON method dev.
Tamas G;
2002
337
Processing of convergent information in identified cortical
networks
OFF domain
Tamas G;Szabadics J;Somogyi P;
2002
1033
Cell type- and subcellular position-dependent summation
of unitary postsynaptic potentials in neocortical neurons
OFF domain
Tammer B;Lehmann I;Nieber K;Altenburger R;
2007
496
Combined effects of mycotoxin mixtures on human T cell
function
ON experimental
study
Tang KY;Lu T;Chang CH;Lo YK;Cheng
JS;Wang JL;Chang HT;Jan CR;
2001
901
Effect of fluoxetine on intracellular Ca2+ levels in bladder
female transitional carcinoma (BFTC) cells
OFF domain
Tapbergenov SO;Koptelov VA;
1981
124
ADRENORECEPTOR-INDEPENDENT ACTION OF
THYROXINE AND ADRENOXYL ON MYOCARDIAL
ADENYLATE-CYCLASE ACTIVITY AND CYCLIC-AMP LEVEL
OFF domain
Taylor FB;Lockhart MS;
1985
207
WHOLE-BLOOD CLOT LYSIS - INVITRO MODULATION BY
ACTIVATED PROTEIN-C
OFF domain
Teixeira MM;Das AM;Miotla JM;Perretti
M;Hellewell PG;
1998
351
The role of lipocortin-1 in the inhibitory action of
dexamethasone on eosinophil trafficking in cutaneous
inflammatory reactions in the mouse
OFF domain
Tetens J;Venugopal CS;Holmes EP;Koch
CE;Hosgood G;Moore RM;
2001
713
In vitro responses of equine colonic arterial and venous
rings to adenosine triphosphate
OFF domain
Teuschler LK;Gennings C;Hartley WR;Carter
H;Thiyagarajah A;Schoeny R;Cubbison C;
2005
1030
The interaction effects of binary mixtures of benzene and
toluene on the developing heart of medaka (Oryzias
latipes)
ON experimental
study
Teuschler LK;Hertzberg RC;
1995
582
Current and future risk assessment guidelines, policy, and
methods development for chemical mixtures
ON review
Tezval H;Merseburger AS;Serth J;Herrmann
TW;Becker JU;Jahn O;Kuczyk MA;
2009
729
Differential Expression of Urocortin in Human Testicular
Germ Cells in Course of Spermatogenesis: Role for
Urocortin in Male Fertility?
OFF domain
Thalhammer O;Havelec L;Knoll E;Wehle E;
1977
60
IQ OF HETEROZYGOTES FOR PHENYLKETONURIA (PKU) INDICATION OF A BLOOD PHENYLALANINEINDEPENDENT ACTION OF PKU MUTANT
OFF domain
Thomas DJ;Wetherhold RC;
1991
370
RELIABILITY-ANALYSIS OF CONTINUOUS FIBER
COMPOSITE LAMINATES
OFF domain
Thomas Y;Sosman J;Irigoyen O;Friedman
SM;Kung PC;Goldstein G;Chess L;
1980
102
FUNCTIONAL-ANALYSIS OF HUMAN T-CELL SUBSETS
DEFINED BY MONOCLONAL-ANTIBODIES .1.
COLLABORATIVE T-T INTERACTIONS IN THE
IMMUNOREGULATION OF B-CELL DIFFERENTIATION
OFF domain
Thompson AK;Mostafapour SP;Denlinger
LC;Bleasdale JE;Fisher SK;
1991
1004
THE AMINOSTEROID U-73122 INHIBITS MUSCARINIC
RECEPTOR SEQUESTRATION AND PHOSPHOINOSITIDE
HYDROLYSIS IN SK-N-SH NEUROBLASTOMA-CELLS - A
ROLE FOR GP IN RECEPTOR COMPARTMENTATION
OFF domain
Thompson MM;
1979
84
GENETICS OF INCOMPATIBILITY IN CORYLUS-AVELLANA
L
OFF domain
APPENDIX A
Authors
Page 49 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Thompson TL;Bridges SR;Weirs WJ;
2001
839
Alteration of dopamine transport in the striatum and
nucleus accumbens of ovariectomized and estrogenprimed rats following N-(p-isothiocyanatophenethyl)
spiperone (NIPS) treatment
OFF domain
Thubagere A;Reinhard BM;
2010
840
Nanoparticle-induced apoptosis propagates through
hydrogen-peroxide-mediated bystander killing: insights
from a human intestinal epithelium in vitro model
OFF domain
Thuillier R;Wang Y;Culty M;
2003
1099
Prenatal exposure to estrogenic compounds alters the
OFF domain
expression pattern of platelet-derived growth factor
receptors alpha and beta in neonatal rat testis:
Identification of gonocytes as targets of estrogen exposure
Thuluvath PJ;Maheshwari A;Mehdi
J;Fairbanks KD;Wu LLW;Gelrud LG;Ryan
MJ;Anania FA;Lobis IF;Black M;
2004
436
Randomised, double blind, placebo controlled trial of
interferon, ribavirin, and amantadine versus interferon,
ribavirin, and placebo in treatment naive patients with
chronic hepatitis C
OFF domain
Thyregod P;Spliid H;
1991
1101
A HYPOTHESIS OF NO INTERACTION IN FACTORIALEXPERIMENTS WITH A BINARY RESPONSE
ON experimental
study
Ting CC;Rodrigues D;
1980
103
IMMUNOREGULATORY CIRCUIT AMONG MACROPHAGE
SUBSETS FOR T-CELL-MEDIATED CYTO-TOXIC
RESPONSE TO TUMOR-CELLS
OFF domain
Tkachenko A;Nesterova L;Pshenichnov M;
2001
782
The role of the natural polyamine putrescine in defense
against oxidative stress in Escherichia coli
OFF domain
Tokimasa T;Ito M;Simmons MA;Schneider
CR;Tanaka T;Nakano T;Akasu T;
1995
352
INHIBITION BY WORTMANNIN OF M-CURRENT IN
BULLFROG SYMPATHETIC NEURONS
OFF domain
Tournier JM;Maouche K;Coraux C;Zahm
JM;Cloez-Tayarani I;Nawrocki-Raby
B;Bonnomet A;Burlet H;Lebargy F;Polette
M;Birembaut P;
2006
706
alpha 3 alpha 5 beta 2-nicotinic acetylcholine receptor
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OFF domain
Treffers HP;Muschel LH;
1954
3
THE COMBINED ACTIONS OF CHLORAMPHENICOL AND
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SALMONELLA-TYPHOSA
OFF domain
Trimble AJ;Belden JB;Mueting SA;Lydy MJ;
2010
468
Determining modifications to bifenthrin toxicity and
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OFF domain
Trimble AJ;Weston DP;Belden JB;Lydy MJ;
2009
735
IDENTIFICATION AND EVALUATION OF PYRETHROID
INSECTICIDE MIXTURES IN URBAN SEDIMENTS
ON experimental
study
Trnka L;Mison P;Staflova S;
1974
33
INTERACTION ASPECTS OF ANTIMYCOBACTERIAL
DRUGS IN CHEMOTHERAPY OF TUBERCULOSIS .2. ROLE
OF RIFAMPICIN AND OTHER DRUGS IN DEPENDENT OR
INDEPENDENT ACTION OF DRUG ASSOCIATIONS
INVITRO
OFF domain
Trottier G;Triggle CR;O'Neill SK;Loutzenhiser
R;
1998
353
Cyclic GMP-dependent and cyclic GMP-independent actions OFF domain
of nitric oxide on the renal afferent arteriole
Tyagi P;Chancellor M;Yoshimura N;Huang L;
2008
1065
Activity of different phospholipids in attenuating
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OFF domain
Ubakov SA;Vanchuova NN;Grebennikov
SA;Frash VN;
1994
1019
Mutagenic action of asbestos, modified by combination
with ethanol, coffee and fumigating resin
ON experimental
study
Uitterlinden EJ;Koevoet JLM;Verkoelen
CF;Bierma-Zeinstra SMA;Jahr H;Weinans
H;Verhaar JAN;van Osch G;
2008
459
Glucosamine increases hyaluronic acid production in
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OFF domain
Uteshev V;
1993
328
THE BINOMIAL-DISTRIBUTION AND THE EVIDENCE FOR
INDEPENDENT ACTION OF ION CHANNELS
OFF domain
V. Dressler;G. Muller;J. Suhnel
1999
1136
CombiTool - A new computer program for analyzing
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ON method dev.
V. Petraitis;R. Petraitiene;W. W. Hope;J.
Meletiadis;D. Mickiene;J. E. Hughes;M. P.
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2009
1151
Combination Therapy in Treatment of Experimental
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ON experimental
study
APPENDIX A
Authors
Page 50 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Van de Meent D;Huijbregts MAJ;
2005
386
Calculating life-cycle assessment effect factors from
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ON method dev.
van Heel DA;Hunt KA;Ghosh S;Herve
M;Playford RJ;
2006
475
Normal responses to specific NOD1-activating
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OFF domain
Vanderkooi JM;Ierokomas A;Nakamura
H;Martonosi A;
1977
61
FLUORESCENCE ENERGY-TRANSFER BETWEEN CA2+
TRANSPORT ATPASE MOLECULES IN ARTIFICIAL
MEMBRANES
OFF domain
Vanhaastert RM;Grote JJ;Vanblitterswijk
CA;Prewett AB;
1994
1056
OSTEOINDUCTION WITHIN PEO/PBT COPOLYMER
IMPLANTS IN CRANIAL DEFECTS USING DEMINERALIZED
BONE-MATRIX
OFF domain
VanIderstine SC;Byers DM;Ridgway ND;Cook
HW;
1997
540
Phospholipase D hydrolysis of plasmalogen and diacyl
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OFF domain
Vasileva-Todorova L;Dipchikova S;Manova
E;Chasonikarova V;
1987
945
EFFECT OF INTENSIVE NOISE AND MICROCLIMATE WITH OFF domain
VARIOUS POWER LEVELS ON SOME CARDIOVASCULAR
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Vasilieva LA;
1984
180
THE ANALYSIS OF THE GENE SYSTEM EXPRESSING THE
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OFF domain
Venard R;Brethes D;Giraud MF;Vaillier
J;Velours J;Haraux F;
2003
628
Investigation of the role and mechanism of IF1 and STF1
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OFF domain
Vercruyssen L;Gonzalez N;Werner
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2011
556
Combining Enhanced Root and Shoot Growth Reveals
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OFF domain
Vermeulen A;Paridaens R;Heuson JC;
1983
161
EFFECTS OF AMINOGLUTETHIMIDE ON ADRENALSTEROID SECRETION
OFF domain
Verro R;Finizio A;Otto S;Vighi M;
2009
587
Predicting Pesticide Environmental Risk in Intensive
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ON experimental
study
Vetvicka V;Lee G;Spitalny GL;Kincade PW;
1988
276
INFLUENCE OF INTERFERON-GAMMA ON B-CELL
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OFF domain
Viarengo A;Bettella E;Fabbri R;Burlando
B;Lafaurie M;
1997
621
Heavy metal inhibition of EROD activity in liver
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ON trivial
Vida I;Halasy K;Szinyei C;Somogyi P;Buhl EH;
1998
358
Unitary IPSPs evoked by interneurons at the stratum
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OFF domain
Vidhu S;Haldar C;
1993
883
Gonad independent action of pineal gland on accessory
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OFF domain
Vighi M;Altenburger R;Arrhenius A;Backhaus
T;Bodeker W;Blanck H;Consolaro F;Faust
M;Finizio A;Froehner K;Gramatica P;Grimme
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2003
982
Water quality objectives for mixtures of toxic chemicals:
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ON review
Villarpalasi C;Kumon A;
1981
125
PURIFICATION AND PROPERTIES OF DOG CARDIAC
TROPONIN-T KINASE
OFF domain
Vilpo J;Koski T;Vilpo L;
2000
437
Calcium antagonists potentiate P-glycoprotein-independent OFF domain
anticancer drugs in chronic lymphocytic leukemia cells in
vitro
Vlakhova M;Lev V;
1980
383
THE EFFECT OF IRRIGATION PLANTING DENSITY AND
HYBRID ON GROWTH DEVELOPMENT AND YIELD OF
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OFF domain
Voss HP;Shukrula S;Wu TS;Donnell D;Bast A;
1994
803
A FUNCTIONAL BETA-2-ADRENOCEPTOR-MEDIATED
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OFF domain
APPENDIX A
Authors
Page 51 of 66
PubYear
ID
CFT/EFSA/PPR/2010/02
Title
Relevance
W. E. Delaney;H. L. Yang;M. D. Miller;C. S.
Gibbs;S. Xiong
2004
1135
Combinations of adefovir with nucleoside analogs produce
additive antiviral effects against hepatitis B virus in vitro
ON experimental
study
W. R. Gould;T. B. McClanahan;K. M. Welch;S.
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2006
1140
Inhibitors of blood coagulation factors Xa and IIa
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ON trivial
W. Y. W. Lee;C. C. M. Cheung;K. W. K. Liu;K.
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2010
1144
Cytotoxic Effects of Tanshinones from Salvia miltiorrhiza
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ON experimental
study
Wahrendorf J;Brown CC;
1980
104
BOOTSTRAPPING A BASIC INEQUALITY IN THE ANALYSIS
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ON trivial
Wallgren A;Festin R;Gidlof C;Dohlsten
M;Kalland T;Totterman TH;
1993
329
EFFICIENT KILLING OF CHRONIC B-LYMPHOCYTIC
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OFF domain
Walsh CJ;Toranto JD;Gilliland CT;Noyes
DR;Bodine AB;Luer CA;
2006
788
Nitric oxide production by nurse shark (Ginglymostoma
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OFF domain
Walsh JV;Singer JJ;
1987
246
IDENTIFICATION AND CHARACTERIZATION OF MAJOR
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OFF domain
Walter H;Consolaro F;Gramatica P;Scholze
M;Altenburger R;
2002
535
Mixture toxicity of priority pollutants at no observed effect
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ON experimental
study
Wanebo HJ;Pace R;Hargett S;Katz D;Sando J;
1986
224
PRODUCTION OF AND RESPONSE TO INTERLEUKIN-2 IN
PERIPHERAL-BLOOD LYMPHOCYTES OF CANCERPATIENTS
OFF domain
Wang C;Liao WP;Wang ML;Lin CC;
2004
507
Miscible blends of syndiotactic polystyrene and atactic
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OFF domain
Wang DM;Masutani H;Oka S;Tanaka
T;Yamaguchi-Iwai Y;Nakamura H;Yodoi J;
2006
681
Control of mitochondrial outer membrane permeabilization
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OFF domain
Wang H;Li Y;Huang H;Xu X;Wang Y;
2011
994
TOXICITY EVALUATION OF SINGLE AND MIXED
ANTIFOULING BIOCIDES USING THE
STRONGYLOCENTROTUS INTERMEDIUS SEA URCHIN
EMBRYO TEST
ON experimental
study
Wang JS;Ho FM;Kang HC;Lin WW;Huang KC;
2011
980
Celecoxib induces heme oxygenase-1 expression in
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Wang LJ;Liu SS;Zhang J;Li WY;
2010
442
A new effect residual ratio (ERR) method for the validation ON method dev.
of the concentration addition and independent action
models
Wang RP;Su JD;Zhang XL;Shi Y;Cui
NR;Onyebuchi VA;Jiang C;
2006
846
Kir6.2 channel gating by intracellular protons: Subunit
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OFF domain
Wang Z;Chen JW;Huang LP;Wang Y;Cai
XY;Qiao XH;Dong YY;
2009
682
Integrated fuzzy concentration addition-independent
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Wang ZQ;Wang HH;Wu JY;Zhu DY;Zhang
XN;Ou LL;Yu YP;Lou YJ;
2009
388
Enhanced co-expression of beta-tubulin III and choline
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OFF domain
Wargovich MJ;Lointier PH;
1987
247
CALCIUM AND VITAMIN-D MODULATE MOUSE COLON
EPITHELIAL PROLIFERATION AND GROWTHCHARACTERISTICS OF A HUMAN-COLON TUMOR-CELL
LINE
OFF domain
Wascher TC;Bachernegg M;Kickenweiz
A;Stark G;Stark U;Toplak H;Graier WF;
1996
908
Involvement of the L-arginine-nitric oxide pathway in
hyperglycaemia-induced coronary artery dysfunction of
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OFF domain
Wati S;Li P;Burrell CJ;Carr JM;
2007
1074
Dengue virus (DV) replication in monocyte-derived
macrophages is not affected by tumor necrosis factor
alpha (TNF-alpha), and DV infection induces altered
responsiveness to TNF-alpha stimulation
OFF domain
APPENDIX A
Authors
Page 52 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Weinberg CR;
1986
225
APPLICABILITY OF THE SIMPLE INDEPENDENT ACTION
MODEL TO EPIDEMIOLOGIC STUDIES INVOLVING 2
FACTORS AND A DICHOTOMOUS OUTCOME
ON trivial
Weinberg M;Kling CL;
1996
399
Uncoordinated agricultural and environmental policy
making: An application to irrigated agriculture in the West
OFF domain
Welbourne TC;Cronin MJ;
1991
896
GROWTH-HORMONE ACCELERATES TUBULAR ACIDSECRETION
OFF domain
Welsh MJ;
1985
208
BASOLATERAL MEMBRANE POTASSIUM CONDUCTANCE
IS INDEPENDENT OF SODIUM-PUMP ACTIVITY AND
MEMBRANE VOLTAGE IN CANINE TRACHEAL EPITHELIUM
OFF domain
Wershow HJ;Ritchey FJ;Alphin TH;
1981
126
PHYSICIANS OPINIONS TOWARD LEGISLATION
DEFINING DEATH AND WITHHOLDING LIFE SUPPORT
OFF domain
Wessinger WD;Evans EB;
1988
277
MODELING MULTIPLE AGENT INTERACTIONS IN
BEHAVIORAL PHARMACOLOGY
ON trivial
Weyerbrock S;Schreieck J;Karch M;Overbeck
M;Meisner H;Kemkes B;Schomig A;Schmitt C;
1997
913
Rate-independent effects of the new class III
antiarrhythmic agent ambasilide on transmembrane action
potentials in human ventricular endomyocardium
OFF domain
Whitehead A, Whitehead J,Todd S, Zhou YH,
Smith MK
2008
1156
Fitting models for the joint action of two drugs using
SAS((R))
ON method dev.
Wieprecht T;Beyermann M;Seelig J;
1999
670
Binding of antibacterial magainin peptides to electrically
neutral membranes: Thermodynamics and structure
OFF domain
Wilde MH;Garvin S;
2007
400
A concept analysis of self-monitoring
OFF domain
Williams DW;Williams ED;Wynfordthomas D;
1988
278
LOSS OF DEPENDENCE ON IGF-1 FOR PROLIFERATION
OF HUMAN THYROID ADENOMA CELLS
OFF domain
Williams TJ;Jose PJ;
1981
127
MEDIATION OF INCREASED VASCULAR-PERMEABILITY
AFTER COMPLEMENT ACTIVATION - HISTAMINEINDEPENDENT ACTION OF RABBIT C5A
OFF domain
Willmore LJ;
1995
672
THE EFFECT OF AGE ON PHARMACOKINETICS OF
ANTIEPILEPTIC DRUGS
OFF domain
Willmott Y;
1997
403
Prison nursing: the tension between custody and care
OFF domain
Wilson WW;Casavant KL;
1991
1014
SOME MARKET POWER IMPLICATIONS OF THE SHIPPING
ACT OF 1984 - A CASE-STUDY OF THE UNITED-STATES
TO PACIFIC RIM TRANSPORTATION MARKETS
OFF domain
Winfield MD;Latifi T;Groisman EA;
2005
624
Transcriptional regulation of the 4-amino-4-deoxy-Larabinose biosynthetic genes in Yersinia pestis
OFF domain
Wise PM;Miyazawa T;Gallagher M;Preti G;
2007
519
Human odor detection of homologous carboxylic acids and
their binary mixtures
ON trivial
Wisniewska A;Wolnicka-Glubisz A;
2004
619
ESR studies on the effect of cholesterol on chlorpromazine
interaction with saturated and unsaturated liposome
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OFF domain
Wolff S;Erster S;Palacios G;Moll UM;
2008
743
p53's mitochondrial translocation and MOMP action is
independent of Puma and Bax and severely disrupts
mitochondrial membrane integrity
OFF domain
Wong ACL;Bergdoll MS;
1990
317
EFFECT OF ENVIRONMENTAL-CONDITIONS ON
PRODUCTION OF TOXIC SHOCK SYNDROME TOXIN-1 BY
STAPHYLOCOCCUS-AUREUS
OFF domain
Wong WT;Nick HS;Frost SC;
1992
888
REGULATION OF ANNEXIN-I IN ADIPOGENESIS - CAMPINDEPENDENT ACTION OF METHYLISOBUTYLXANTHINE
OFF domain
Wright JR;Colby HD;Miles PR;
1980
105
LIPID-PEROXIDATION IN GUINEA-PIG LUNG
MICROSOMES
OFF domain
Wu PF;Chang LS;Kao YL;Wang KT;
2008
618
beta-Bungarotoxin induction of neurite outgrowth in
NB41A3 cells
OFF domain
Xiao SH;Keiser J;Chollet J;Utzinger J;Dong
YX;Endriss Y;Vennerstrom JL;Tanner M;
2007
809
In vitro and in vivo activities of synthetic trioxolanes
against major human schistosome species
OFF domain
APPENDIX A
Authors
Page 53 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Xie FL;Koziar SA;Lampi MA;Dixon
DG;Norwood WP;Borgmann U;Huang
XD;Greenberg BM;
2006
759
Assessment of the toxicity of mixtures of copper, 9,10phenanthrenequinone, and phenanthrene to Daphnia
magna: Evidence for a reactive oxygen mechanism
ON experimental
study
Xu QJ;Li SH;Zhao Y;Maures TJ;Yin P;Duan
CM;
2004
630
Evidence that IGF binding protein-5 functions as a ligandindependent transcriptional regulator in vascular smooth
muscle cells
OFF domain
Xu X;Li Y;Wang YA;Wang YH;
2011
1025
Assessment of toxic interactions of heavy metals in multicomponent mixtures using sea urchin embryo-larval
bioassay
ON experimental
study
Xu XY;Zhu YW;Wang BC;Shen XQ;
2003
975
Surface modification of nanodiamond in aqueous medium
OFF domain
Yabe N;Komiya K;Takezono T;Matsui H;
1993
668
LYSOZYME AS A REGULATOR OF INTERLEUKIN-2ACTIVATED LYMPHOCYTE-PROLIFERATION
OFF domain
Yager JD;Campbell HA;Longnecker
DS;Roebuck BD;Benoit MC;
1984
181
ENHANCEMENT OF HEPATOCARCINOGENESIS IN FEMALE
RATS BY ETHINYL ESTRADIOL AND MESTRANOL BUT
NOT ESTRADIOL
ON trivial
Yajima H;Komatsu M;Schermerhorn T;Aizawa
T;Kaneko T;Nagai M;Sharp GWG;Hashizume
K;
1999
511
CAMP enhances insulin secretion by an action on the ATPsensitive K+ channel-independent pathway of glucose
signaling in rat pancreatic islets
OFF domain
Yajima Y;Akita Y;Saito T;
1986
226
PERTUSSIS TOXIN BLOCKS THE INHIBITORY EFFECTS OF OFF domain
SOMATOSTATIN ON CAMP-DEPENDENT VASOACTIVEINTESTINAL-PEPTIDE AND CAMP-INDEPENDENT
THYROTROPIN RELEASING HORMONE-STIMULATED
PROLACTIN SECRETION OF GH3 CELLS
Yamamoto JK;Blalock JE;Johnson HM;
1982
146
HUMAN NATURAL KILLER-LIKE ACTIVITY AGAINST
MOUSE SPLEEN-CELLS
OFF domain
Yamamoto K;Takeuchi K;Ohishi M;Theodore
K;Rakugi H;
2010
348
PPAR gamma Dependent and Independent Action of
Telmisartan in Energy Production, Oxidative Stress and
Apoptosis; Possible Role in Mitochondrial Function
OFF domain
Yamauchi Y;Okazaki H;Desaki M;Kohyama
T;Kawasaki S;Yamamoto K;Takizawa H;
2004
680
Methotrexate induces interleukin-8 production by human
bronchial and alveolar epithelial cells
OFF domain
Yamazaki K;Yamamoto T;Kawai Y;Inoue N;
2004
752
Enhancement of antilisterial activity of essential oil
constituents by nisin and diglycerol fatty acid ester
OFF domain
Yang ZH;
1982
147
EXPANSION OF THE ENTERPRISES RIGHT TO
OFF domain
INDEPENDENT ACTION SETS NEW AGENDA FOR HIGHEREDUCATION
Yap J;Tsao T;Fawcett J;Fielder PJ;Keller
GA;Rabkin R;
1997
953
Effect of insulin-like growth factor binding proteins on the
response of proximal tubular cells to insulin-like growth
factor-I
OFF domain
Yau L;Wilson DP;Werner JP;Zahradka P;
2001
477
Bradykinin receptor antagonists attenuate neointimal
proliferation postangioplasty
OFF domain
Yeh SW;Wei LS;Nelson AI;Steinberg MP;
1982
148
FREEZE-THAW STABILITY OF ILLINOIS SOYBEAN
BEVERAGE
OFF domain
Yeoman RR;Jones WD;Rizk BM;
1998
643
Evidence for nitric oxide regulation of hamster sperm
hyperactivation
OFF domain
Yoshikawa M;Motoshima K;Fujimoto K;Tai
A;Kakuta H;Sasaki K;
2008
671
Pyridinium cationic-dimer antimalarials, unlike chloroquine,
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stage of Plasmodium falciparum
OFF domain
Yoshimi T;Hashimoto F;Takahashi
S;Takahashi Y;
2010
1010
Suppression of embryonic lung branching morphogenesis
by antisense oligonucleotides against HOM/C homeobox
factors
OFF domain
Yu GX;Park BH;Chandramohan P;Munavalli
R;Geist A;Samatova NF;
2005
869
In silico discovery of enzyme-substrate specificitydetermining residue clusters
OFF domain
Yu QJ;Zhou N;Bian JF;Hao XK;Wang YM;Guo
WY;Wang HC;Gao F;
2008
343
Insulin-titrated Maintenance of Normoglycemia and
Glycemia-independent cardioprotection in
Ischemic/Reperfused Dogs
OFF domain
Yuao Zhu;Maria Curtis;Xiaoping Qi;Michael D.
Miller;Katyna Borroto-Esoda
2009
1159
Anti-hepatitis B virus activity in vitro of combinations of
tenofovir with nucleoside/nucleotide analogues
ON experimental
study
APPENDIX A
Authors
Page 54 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Zagotta WN;Aldrich RW;
1990
318
VOLTAGE-DEPENDENT GATING OF SHAKER A-TYPE
POTASSIUM CHANNELS IN DROSOPHILA MUSCLE
OFF domain
Zaidi M;Davies TF;Zallone A;Blair HC;Iqbal
J;Moonga SS;Mechanick J;Sun L;
2009
649
Thyroid-stimulating hormone, thyroid hormones, and bone
loss
OFF domain
Zamoner A;Bruno AN;Casali EA;Corbelini
PF;Diniz GP;Barreto-Chaves MLM;Silva
F;Sarkis JJF;Pessoa-Pureur R;
2006
1013
Genomic-independent action of thyroid hormones on
NTPDase activities in Sertoli cell cultures from congenital
hypothyroid rats
OFF domain
Zamrazil V;Nemec J;Neradilova M;Nedvidkova
J;Blahosova A;Felt V;Havelka J;Bednar J;
1981
128
COMPARISON OF METABOLIC EFFECTS OF GLUCAGON
OFF domain
AND CALCITONIN AND ASSESSMENT OF DIRECT EFFECT
OF GLUCAGON ON CALCITONIN LEVEL IN ATHYROID MAN
Zanfolin M;Faro R;Araujo EG;Guaraldo
AMA;Antunes E;De Nucci G;
2006
1053
Protective effects of BAY 41-2272 (sGC stimulator) on
hypertension, heart, and cardiomyocyte hypertrophy
induced by chronic L-NAME treatment in rats
OFF domain
Zangar RC;Fan YY;Chapkin RS;
2004
767
Interactions of phospholipase D and cytochrome P450
protein stability
OFF domain
Zaroogian G;Voyer RA;
1995
1070
INTERACTIVE CYTOTOXICITIES OF SELECTED ORGANIC
AND INORGANIC SUBSTANCES TO BROWN CELLS OF
MERCENARIA-MERCENARIA
OFF domain
Zaug AJ;Grosshans CA;Cech TR;
1988
279
SEQUENCE-SPECIFIC ENDORIBONUCLEASE ACTIVITY OF
THE TETRAHYMENA RIBOZYME - ENHANCED CLEAVAGE
OF CERTAIN OLIGONUCLEOTIDE SUBSTRATES THAT
FORM MISMATCHED RIBOZYME SUBSTRATE COMPLEXES
OFF domain
Zecca M;Pession A;Messina C;Bonetti F;Favre
C;Prete A;Cesaro S;Porta F;Mazzarino
I;Giorgiani G;Rondelli R;Locatelli F;
1999
777
Total body irradiation, thiotepa, and cyclophosphamide as
a conditioning regimen for children with acute
lymphoblastic leukemia in first or second remission
undergoing bone marrow transplantation with HLAidentical siblings
OFF domain
Zecevic D;Antic S;
1998
609
Fast optical measurement of membrane potential changes
at multiple sites on an individual nerve cell
OFF domain
Zedginidze MS;Spirande IV;Nikonenko
BV;Fuks BB;
1982
435
MUTUAL SUPPRESSION OF NATURAL KILLERS AND
TUMOR CELLS THE ROLE OF HOMOLOGY IN THE IC
SUBREGION OF H-2 COMPLEX
OFF domain
Zhang H;Kong FX;Wang SH;Yu Y;Zhang M;
2009
1020
Vitellogenin Induction by a Mixture of Steroidal Estrogens
in Freshwater Fishes and Relevant Risk Assessment
ON experimental
study
Zhang H;Kong Fx;Wang Sh;Yu Y;Zhang
M;Chen Mj;Tan X;Qian Sq;
2008
497
Mixture effects to vitellogenin induction by four
environmental estrogens in freshwater fish
ON experimental
study
Zhang H;Kong Fx;Yu Y;Shi XL;Zhang M;Tian
HE;
2010
974
Assessing the combination effects of environmental
estrogens in fish
ON experimental
study
Zhang H;Patel SA;Kandil E;Mueller CM;Lin
YY;Zenilman ME;
2003
444
Pancreatic elastase is proven to be a mannose-binding
protein - implications for the systemic response to
pancreatitis
OFF domain
Zhang Hc;Chen Ly;Liu Ss;Yin Dq;
2009
876
Jointed Estrogenic Activities of Bisphenol A and Three of
Its Analogs
ON experimental
study
Zhang J;Liu Ss;Dou RN;Liu Hl;Zhang J;
2011
645
Evaluation on the toxicity of ionic liquid mixture with
antagonism and synergism to Vibrio qinghaiensis sp.-Q67
ON experimental
study
Zhang J;Wang Jc;Liu Ss;Ge Hl;Liu Hl;
2009
646
Study on the Toxicity Interaction between Dimethoate and
C16H31ClN2 by Central Composite Design with the Fixed
Concentration Ratio Ray
ON experimental
study
Zhang LQ;Rasenick MM;
2010
494
Chronic Treatment with Escitalopram but Not R-Citalopram OFF domain
Translocates G alpha(s) from Lipid Raft Domains and
Potentiates Adenylyl Cyclase: A 5-Hydroxytryptamine
Transporter-Independent Action of This Antidepressant
Compound
Zhang MIN;Oneil RG;
1996
1092
A regulated calcium channel in apical membranes of renal
proximal tubule cells
OFF domain
Zhang Sheng YS;Lin H;
1995
481
Gastric cancer with P-53 overexpression and nm23 lowexpression has high potential for lymph node metastasis
OFF domain
APPENDIX A
Authors
Page 55 of 66
PubYear
CFT/EFSA/PPR/2010/02
ID
Title
Relevance
Zhang YD;Tazzeo T;Chu V;Janssen LJ;
2006
722
Membrane potassium currents in human radial artery and
their regulation by nitric oxide donor
OFF domain
Zhang YH;Liu SS;Liu HL;Liu ZZ;
2010
455
Evaluation of the combined toxicity of 15 pesticides by
uniform design
ON experimental
study
Zhang YH;Liu SS;Song XQ;Ge HL;
2008
736
Prediction for the mixture toxicity of six organophosphorus
pesticides to the luminescent bacterium Q67
ON experimental
study
Zhang YX;Soto J;Park K;Viswanath G;Kuwada
S;Abel ED;Wang L;
2010
817
Nuclear Receptor SHP, a Death Receptor That Targets
Mitochondria, Induces Apoptosis and Inhibits Tumor
Growth
OFF domain
Zheng YF;Ao ZJ;Jayappa KD;Yao XJ;
2010
454
Characterization of the HIV-1 integrase chromatin-and
LEDGF/p75-binding abilities by mutagenic analysis within
the catalytic core domain of integrase
OFF domain
Zhong Y;Zhou J;Li Y;Lu L;Clemmons;Duan C;
2010
349
IGF Binding Protein-3 Exerts Its IGF-Independent Action
by Antagonizing BMP Action In Vivo
OFF domain
Zhou HL;Li XM;Meinkoth J;Pittman RN;
2000
749
Akt regulates cell survival and apoptosis at a
postmitochondrial level
OFF domain
Zhou XF;Sang WJ;Liu SS;Zhang YL;Ge HL;
2010
590
Modeling and prediction for the acute toxicity of pesticide
mixtures to the freshwater luminescent bacterium Vibrio
qinghaiensis sp.-Q67
ON experimental
study
Zhou ZX;Wang LP;Song ZY;Saari JT;McClain
CJ;Kang YJ;
2004
532
Abrogation of nuclear factor-kappa B activation is involved
in zinc inhibition of lipopolysaccharide-induced tumor
necrosis factor-alpha production and liver injury
OFF domain
Ziemlanski S;Wielgus-Serafinska E;PanczenkoKresowska B;Zelakiewicz K;
1984
588
EFFECT OF LONG-TERM DIET ENRICHMENT WITH
SELENIUM VITAMIN E AND VITAMIN B-15 ON THE
DEGREE OF FATTY INFILTRATION OF THE LIVER
OFF domain
Zouki C;Beauchamp M;Baron C;Filep JG;
1997
483
Prevention of in vitro neutrophil adhesion to endothelial
cells through shedding of L-selectin by C-reactive protein
and peptides derived from C-reactive protein
OFF domain
Zwadloklarwasser G;Platen S;Schmutzler W;
1994
819
INHIBITION OF SPONTANEOUS AND MITOGEN-INDUCED
LYMPHOCYTE-PROLIFERATION BY MURINE BONEMARROW-DERIVED MACROPHAGES - ROLE OF
PROSTAGLANDINS, NITRIC OXIDES AND CELL-TO-CELL
CONTACT
OFF domain
Zwart MP;Hemerik L;Cory JS;de Visser
J;Bianchi F;Van Oers MM;Vlak JM;Hoekstra
RF;Van der Werf W;
2009
836
An experimental test of the independent action hypothesis
in virus-insect pathosystems
OFF domain
Chemical name
Number of studies chemical was included in
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
CFT/EFSA/PPR/2010/02
Acetaldehyde
acetonitrile
acifluorfen
Aniline
Asulam
benzyl butyl phthalate
Carbofuran
Cobalt
DEHP
diclofenac
diisobutyl phthalate
DTMAC
Endosulfan
ethinylestradiol
Fenfuram
flumequine
Fluometuron
Formaldehyde
fusidic acid
Hexachlorobutadiene
Kresoxim-methyl
lambda-cyhalothrin
lead
mecoprop
mesotrion
Metolachlor
Norflurazon
oxolinic acid
p,p'-DDE
p,p'-DDT
PCB-153
phenanthrene
pirimicarb
Prometron
Prometryn
streptomycin
TCDD
Triadimenol
Tributyltin chloride
valproic acid
β-HCH
1,2,3,4-tetrachlorobenzene
1,2,3-Trichlorobenzene
1,3-Dichlorobenzene
1-octanol
1-PCDF
2,3,5-Trichlorophenol
2,3-Dichloropropene
2,4,5-Trichloroaniline
2,4,5-trichlorophenol
2,4-Dichloroaniline
2,4-dinitroaniline
2,5-Dichloroaniline
2-Chloroaniline
3-benzylidene camphor
3-Chloroaniline
3-Chlorophenol
4-chloro-3-methylphenol
4-chlorophenol
Page 56 of 66
12
12
11
11
10
10
8
8
8
8
7
7
7
7
7
6
6
6
6
6
6
6
6
6
5
5
5
5
5
5
5
5
5
5
5
5
5
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
3
3
3
3
APPENDIX B
copper
nickel
Cadmium
Chlorpyrifos
Estradiol
Terbuthylazine
diuron
imidacloprid
Phenol
vinclozolin
Atrazine
Dimethoate
prochloraz
Simetryn
zinc
2,4-Dichlorophenol
2-Chlorophenol
Bisphenol A
Dichlorvos
diquat
genistein
glyphosate
Malathion
Nalidixic acid
2,4,6-trichlorophenol
2,4-dinitrophenol
3,4-Dichloroaniline
8-azaguanine
Benzene
Chloramphenicol
dibutyl phthalate
Fluoranthene
Isoproturon
Linuron
Metsulfuron-methyl
procymidone
Toluene
4-nitrophenol
Aclonifen
Azaserine
bromacil
Cadmium chloride
CCCP
diazinon
diethylhexyl phthalate
Metalaxyl
Metazachlor
Methoxychlor
methyl-parathion
Naphthalene
o,p'-DDT
Paraquat dichloride
Pentachlorophenol
thiacloprid
2,4-D
2-phenylphenol
4-Chloroaniline
4-tert-octylphenol
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
CFT/EFSA/PPR/2010/02
Itraconazole
Lindane
lomefloxacin
Loperamide
Lysinoalanine
malononitrile
MCPA
metamitron
Methoprotryne
methyl methanesulfonate
Metobromuron
Metoxuron
Mirex
monocrothophos
Monolinuron
Monuron
m-Xylene
naringenin
nitrobenzene
nonylphenol
norfloxacin
OCDF
octylphenol
ofloxacin
Parathion
PCB-101
PCB-105
PCB-126
PCB-138
PCB-156
PCB-169
PCB-180
PCB-28
PCB-52
PCB-77
PCDD
pendimethaline
Pentachlorobenzene
Pentanoic acid
permethrin
phosphamidon
pipemidic acid
piromidic acid
Propazine
Propiconazole
propranolol
pyrene
quercetin
resorcinol
retinoic acid
Sea-Nine
Sebuthylazine
Secbumeton
sertraline
Simazine
Stannous chloride
TCDF
Terbutryn
triasulfuron
Page 57 of 66
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
APPENDIX B
4-nonylphenol
4-PCDF
4-phenylazophenol
5-fluoruracil
Acetamiprid
acrolein
acrylamide
actinomycin
Alachlor
Ametryn
amphotericin B
ampicillin
Arochlor1254
Atraton
benzophenone-1
benzophenone-2
Benzylchloride
Buturon
butyraldehyde
Butyric acid
carbamazepine
carbaryl
catechin
catechol
Chlorbromuron
Chlorfenvinphos
chlorothalonil
Chlortoluron
chromium
cinoxacin
coumestrol
Cyanazine
cycloserine
deltamethrin
Desmetryn
Dicamba
dichloromethane
Diltiazem
Dimethametryn
di-n-butyl phthalate
dipentyl phthalate
Dipropetryn
emtricitabine
enoxacin
epicatechin
esfenvalerate
Ethacrynic acid
ethylbenzene
Fenitrothion
Fenuron
flutamide
glutaraldehyde
Heptachlor
hydroquinone
ibuprofen
Irgarol
Iron
isofenphos
isoxaflutole
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Acetochlor
acetone
acetophenone
acrylonitrile
Adefovir
aldicarb
Aldrin
Allyl alcohol
Allylchloride
alpha-chaconine
alpha-cypermethrin
alpha-naphthoflavone
alpha-solanine
amdoxovir
ammonia
anidulafungin
anthracene
apigenin
Arsenic
ascorbic acid
Aspirin
avermectin
azadirachtin
azinphos-methyl
azothymine
azoxystrobin
benomyl
bentazone
benz[a]anthracene
benzo[ghi]fluoranthene
Benzophenone
benzophenone-3
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
CFT/EFSA/PPR/2010/02
3-trifluormethyl-4-nitrophenol
4,4'-dihydroxybenzophenone
4,6-dinitro-o-cresol
4-aminonitrobenzene
4-bromoaniline
4-chloro-3-methylnitrobenzene
4-chlorobenzenethiol
4-chloronitrobenzene
4-Chlorotoluene
4-ethylphenol
4-hydroxybenzaldehyde
4-methyl-2-pentanone
4-methylnitrobenzene
4-nitrobenzyl chloride
4-n-octylphenol
4-n-pentylphenol
4-propylphenol
4-pyridinecarboxaldehyde
4-tert-nonylphenol
5-chloro-2-aminonitrobenzene
6-aminonicotinamide
6-chloro-2,3-dihydrobenzoxazol-2-one
9,10-phenanthrenequinone
a,a'-Dichloro-m-xylene
Acephate
acetic acid
Page 58 of 66
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
APPENDIX B
Trichlorfon
trichloroethylene
velpar
Voriconazole
1,2,4-Trichlorobenzene
1,2,4-trimethylbenzene
1,2-epoxybutane
1,3-dimethylnaphthalene
1,4-Dichloro-2-butene
1-Chloro-2,4-dinitrobenzene
1-hexanol
1-methylnaphthalene
1-napthol
1-nonanol
1-pentanol
2,3,4,5-tetrachloroaniline
2,3,4,5-Tetrachlorophenol
2,3,4,6-tetrachlorophenol
2,3,4-Trichloroaniline
2,3,4-trichlorophenol
2,3,5,6-Tetrachlorophenol
2,3,6-Trichlorophenol
2,3,6-trinitrophenol
2,3-epoxypropyl benzene
2,4,5-trichloronitrobenzene
2,4,6-trichloroaniline
2,4,6-trinitrophenol
2,4-a-Trichlorotoluene
2,4-Dichlorotoluene
2,4-dimethylphenol
2,4-dinitro-1-napthol
2,6-dinitro-4-methyl-phenol
2,6-dinitrophenol
2-butyn-1,4-diol
2-chloronitrobenzene
2-ethylhexanoic acid
2-hydroxybenzaldehyde
2-hydroxyethylacrylate
2-methylnaphthalene
2-nitrophenol
2-nitrotoluene
2-phenylnaphthalene
2-pyridinecarboxaldehyde
3,4,5-Trichlorophenol
3,4-dinitrophenol
3,5-Dichloroaniline
3,5-Dichlorophenol
3-aminonitrobenzene
3-butoxyethanol
3-chloronitrobenzene
3-hydroxybenzaldehyde
3-methoxyphenol
3-methyl-4-aminonitrobenzene
3-methylbenzyl chloride
3-nitroaniline
3-nitrochlorobenzene
3-nitrophenol
3-nitrotoluene
3-pyridinecarboxaldehyde
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
CFT/EFSA/PPR/2010/02
diflubenzuron
dihydrotanshinone
diisododecyl phthalate
diisoheptyl phthalate
diisononyl phthalate
Dimethachlor
dinitramine
Di-n-octyltin dichloride
dinoserb
dinoterb
dioctyl phthalate
d-limonene
DMBIB
dodecylpyridinium bromide
dodine
DPB
Dylox
Edifenphos
entecavir
epichlorohydrin
epoxiconazole
erythromycin
estragole
estriol
estrone
ethofumesate
ethyl 2--4-hydroxyphenoxy propanoate
ethyl acetate
ethyl-4-aminobenzoate
ethylacrylate
ethylparaben
ethyl-parathion
Etrimfos
eugenol
famphur
fatty acid methyl ester mixture
FCCP
Fenarimol
fenobucarb
fenoxaprop-p-ethyl
fenpropidin
fenpropimorph
finasteride
florfenicol
fluoxetine
fosthiazate
gamma-eudesmol
geraniol
gliotoxin
glutardialdehyde
gold nanoparticles
halosulfuron methyl
HCV796
hexabromobisphenol A
hexabromocyclododecane
Hexachlorobenzene
Hexachlorocyclohexane
hexazinone
hydrochloric acid
Page 59 of 66
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
APPENDIX B
Benzyl n-butyl phthalate
benzyl salicylate
BHA
bifenthrin
Biphenyl
bis-2-ethylhexyl adipate
bromoxynil
Butachlor
Butamifos
butylbenzyl phthalate
C10 butyl ester sulfonate
C11.8 LAS
C12 amyl ester sulfonate
C12 butyl ester sulfonate
C13 methyl ester sulfonate
C14 butyl ester sulfonate
C14 methyl ester sulfonate
C7 heptyl ester sulfonate
C8 hexyl ester sulfonate
C8 sec hexyl ester sulfonate
captan
captopril
carbendazim
carbosulfan
carvacrol
Chloral hydrate
chloridazon
chlormephos
Chloroacetone
chlorobenzene
Chloroform
chlortetracycline
citalopram
citric acid
citrinin
Clemizole
clevudine
clofibrinic acid
clopyralid
copper pyrithione
cryptotanshinone
cycloxydim
cyfluthrin
cypermethrin
cyprodinil
CytoSol
daidzein
DCPMU
DE71
decylamine
di n-butyl phthalate
dibenzo[a,l]pyrene
dibenzothiopene
dicapthon
dichlofluanid
Dieldrin
diethyl phthalate
diethylene glycol
diethylstilbestrol
1
1
1
1
1
1
ionic
ionic
ionic
ionic
ionic
ionic
ionic
1
1
1
1
1
1
1
liquid
liquid
liquid
liquid
liquid
liquid
liquid
bmimC8OS
bmimCl
bmimCOS
C11H13BF4N2(S7)
C11H13CIN2(S8)
C12H23BrN2(S9)
C14H23BF4N2(S10)
CFT/EFSA/PPR/2010/02
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Page 60 of 66
ionic liquid C14H27ClN(S11)
ionic liquid C16H31CIN2
ionic liquid C16H31CIN2(S12)
ionic liquid C6H11BF4N2(S1)
ionic liquid C8H15BF4N2 (S3)
ionic liquid C8H15CIN2 (S2)
ionic liquid C9H14BF4N(S4)
ionic liquid C9H17BF4N2(S5)
ionic liquid C9H17BrN2(S6)
ionic liquid emimCl
ionic liquid hmimCl
ionic liquid IM14 (CF3SO2)2N
ionic liquid IM14 BF4
ionic liquid IM18 BF4
ionic liquid omim CO3S
ionic liquid omimC8OS
ionic liquid omimCl
ioxynil
Iprobenphos
iprodione
irgarol 1051
irgasan
isobutylacrylate
isoniazid
Isoxathion
ketoconazole
lactonitrile
lamivudine
L-carvone
Lead chloride
lenacil
letrozole
light Libyan crude petroleum
linear alkyl benzene sulfonate
Magnesium
mancozeb
manganese
Mercuric chloride
mercury
metalaxyl-M
Metaldehyde
metformin
metham sodium
methanol
Methidathion
metoprolol
metribuzin
Miconazole
mitomycin c
Monochlorobenzene
musk ketone
naproxen
n-butanol
nicotine
NM283
N-methyl-N-nitrosourea
nonylamine
N-phenyl-2-napthylamine
ochratoxin A
octanol
oseltamivir acid
oseltamivir ethylester
o-Tolualdehyde
o-xylene
Oxytetracycline
paroxetine
patulin
PBDE-100
PBDE-209
PBDE-47
PBDE-99
PCB118
PCB-118
p-Cresol
p-Dimethoxybenzene
pentachloroethane
Pentachloropyridine
phenyl salicylate
Phoxim
piperonyl butoxide
Pirimiphos-methyl
p-nitroso-N,N-dimethylaniline
p-Octylphenol
potassium citrate
Potassium cyanide
potassium dichromate
Potassium nitrite
p-phenoxyphenol
Pretilachlor
prometon
pronamide
Propachlor
Propargyl alcohol
propionaldehyde
propylparaben
Prothiophos
p-tert-Pentylphenol
Pulegone
Pyraclofos
Ravuconazole
retene
ribavarin
rifampicin
APPENDIX B
hydrogen peroxide
hydroxylamine
hydroxyurea
imazalil
interferon
ionic liquid bmimBr
Page 61 of 66
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
APPENDIX B
SCH503034
Sea-nine 211
selenium
semicarbazide
silver nanoparticles
sodium azide
sodium citrate
Sodium metabisulphite
Spermine
spirodiclofen
styrene oxide
sucrose
sulfamethoxazole
tamoxifen
tanshinone I
tanshinone IIA
tebuconazole
telbivudine
Tenofovir
terbacil
Terbinafine
terbumeton
Testosterone
tetrabromobisphenol A
tetrahydrofuran
thifensulfuron methyl
thiram
Thymol
Tolclofos-methyl
tolylfluanid
trans-1,4-dichloro-2-butene
trans-anethole
tribenuron methy;
Triphenyltin chloride
Turbumeton
Vaelpar
Vamidothion
Vanadium
VX950
zinc pyrithione
CFT/EFSA/PPR/2010/02
APPENDIX C
Page 62 of 66
CFT/EFSA/PPR/2010/02
List of chemicals tested in 'dual relevance' mixture studies
Chemical name
Mode/mechanism, if stated
Mixture study: First
author, Pub. year
ID
Che Mix Art#
m# #
Aclonifen
porphyrin biosynthesis
Faust , 2003
1
2
1 1
8-azaguanine
RNA synthesis and function
Faust , 2003
3
3
1 1
Azaserine
Nucleotide biosynthesis
Faust , 2003
4
4
1 1
CCCP
Proton translocation and ATP
synthesis
Faust , 2003
5
5
1 1
Chloramphenicol
protein biosynthesis
Faust , 2003
6
6
1 1
DTMAC
Membrane functions
Faust , 2003
7
7
1 1
Fenfuram
Respiratory electron transport
Faust , 2003
8
8
1 1
Kresoxim-methyl
Respiratory electron transport
Faust , 2003
9
9
1 1
Metalaxyl
RNA synthesis and function
Faust , 2003
10
10
1 1
Metazachlor
lipid biosynthesis
Faust , 2003
11
11
1 1
Metsulfuron-methyl
amino acid biosynthesis
Faust , 2003
12
12
1 1
Nalidixic acid
DNA synthesis and function
Faust , 2003
13
13
1 1
Norflurazon
Carotenoid biosynthesis
Faust , 2003
14
14
1 1
Paraquat dichloride
Photosynthetic electron transport Faust , 2003
15
15
1 1
Terbuthylazine
Photosynthetic electron transport Faust , 2003
16
16
1 1
Triadimenol
steroid biosynthesis
Faust , 2003
17
17
1 1
Atrazine
photosystem II inhibitor
Walter , 2002
62
20
2 11
Biphenyl
not known
Walter , 2002
63
21
2 11
Chloral hydrate
not known
Walter , 2002
64
22
2 11
2,4,5-trichlorophenol
not known
Walter , 2002
65
23
2 11
Fluoranthene
not known
Walter , 2002
66
24
2 11
Lindane
not known
Walter , 2002
67
25
2 11
Naphthalene
not known
Walter , 2002
68
26
2 11
Parathion
not known
Walter , 2002
69
27
2 11
Phoxim
not known
Walter , 2002
70
28
2 11
Tributyltin chloride
inhibition of photosynthesis
Walter , 2002
71
29
2 11
Triphenyltin chloride
inhibition of photosynthesis
Walter , 2002
72
30
2 11
vinclozolin
not stated
Eustache , 2009
36
31
12 60
DEHP
Groten , 1997
97
34
18 15
1,3-Dichlorobenzene
Hermens , 1985
128
38
5 9
1,2,3-Trichlorobenzene
Hermens , 1985
129
39
5 9
1,2,3-Trichlorobenzene
Wade , 2002
83
39
17 16
Monochlorobenzene
Hermens , 1985
130
40
5 9
1,2,3,4-tetrachlorobenzene
Hermens , 1985
131
41
5 9
APPENDIX C
Chemical name
Page 63 of 66
Mode/mechanism, if stated
1,2,3,4-tetrachlorobenzene
Benzene
Nonpolar narcotic
Mixture study: First
author, Pub. year
Che Mix Art#
m# #
85
41
17 16
Wang , 2009
23
42
11 71
132
42
5 9
86
43
17 16
133
43
5 9
24
44
11 71
Hermens , 1985
Pentachlorobenzene
Wade , 2002
Pentachlorobenzene
Hermens , 1985
Nonpolar narcotic
ID
Wade , 2002
Benzene
Toluene
CFT/EFSA/PPR/2010/02
Wang , 2009
Toluene
Hermens , 1985
134
44
5 9
m-Xylene
Hermens , 1985
136
45
5 9
4-Chlorotoluene
Hermens , 1985
137
46
5 9
Chloroform
Hermens , 1985
138
47
5 9
2,4-Dichlorotoluene
Hermens , 1985
135
48
5 9
2,4-Dichlorotoluene
Hermens , 1985
147
48
5 9
2-Chloroaniline
Hermens , 1985
139
49
5 9
3,5-Dichloroaniline
Hermens , 1985
140
50
5 9
2,3,4-Trichloroaniline
Hermens , 1985
141
51
5 9
3,4-Dichloroaniline
Hermens , 1985
142
52
5 9
Aniline
Hermens , 1985
143
53
5 9
27
53
11 71
Aniline
Polar narcotic
Wang , 2009
3-Chloroaniline
Hermens , 1985
144
54
5 9
4-Chloroaniline
Hermens , 1985
145
55
5 9
2,5-Dichloroaniline
Hermens , 1985
146
56
5 9
2,4,5-Trichloroaniline
Hermens , 1985
148
58
5 9
2,3,4,5-tetrachloroaniline
Hermens , 1985
149
59
5 9
3-Chlorophenol
Hermens , 1985
150
60
5 9
30
61
11 71
2,4-Dichlorophenol
Polar narcotic
Wang , 2009
2,4-Dichlorophenol
Hermens , 1985
151
61
5 9
3,4,5-Trichlorophenol
Hermens , 1985
152
62
5 9
2,3,5,6-Tetrachlorophenol
Hermens , 1985
153
63
5 9
26
64
11 71
Phenol
Polar narcotic
Wang , 2009
Phenol
Hermens , 1985
154
64
5 9
2-Chlorophenol
Hermens , 1985
155
65
5 9
3,5-Dichlorophenol
Hermens , 1985
156
66
5 9
2,3,5-Trichlorophenol
Hermens , 1985
157
67
5 9
2,3,6-Trichlorophenol
Hermens , 1985
158
68
5 9
2,3,4,5-Tetrachlorophenol
Hermens , 1985
159
69
5 9
Pentachlorophenol
Hermens , 1985
160
70
5 9
Broderius , 2005
58
79
6 118
Hexachlorobutadiene
reactive toxicant
APPENDIX C
Chemical name
Page 64 of 66
Mode/mechanism, if stated
CFT/EFSA/PPR/2010/02
Mixture study: First
author, Pub. year
ID
Che Mix Art#
m# #
Sodium metabisulphite
Jonker , 1990
120
80
20 12
Mirex
Wade , 2002
81
81
17 16
Mirex
Jonker , 1990
121
81
20 12
Loperamide
Groten , 1997
94
82
18 15
Loperamide
Jonker , 1990
122
82
20 12
Metaldehyde
Jonker , 1990
123
83
20 12
Di-n-octyltin dichloride
Jonker , 1990
124
84
20 12
Stannous chloride
Groten , 1997
93
85
18 15
Stannous chloride
Jonker , 1990
125
85
20 12
Lysinoalanine
Jonker , 1990
126
86
20 12
Potassium nitrite
Jonker , 1990
127
87
20 12
Acephate
Ito , 1995
100
90
19 14
Butamifos
Ito , 1995
101
91
19 14
Chlorfenvinphos
Ito , 1995
102
92
19 14
Chlorpyrifos
Broerse , 2010
22
93
Chlorpyrifos
Ito , 1995
103
93
19 14
Dichlorvos
Ito , 1995
104
94
19 14
Dimethoate
Ito , 1995
105
95
19 14
Edifenphos
Ito , 1995
106
96
19 14
Endosulfan
Ito , 1995
107
97
19 14
Endosulfan
Wade , 2002
77
97
17 16
Etrimfos
Ito , 1995
108
98
19 14
Fenitrothion
Ito , 1995
109
99
19 14
Iprobenphos
Ito , 1995
110 100
19 14
Isoxathion
Ito , 1995
111 101
19 14
Malathion
Ito , 1995
112 102
19 14
Methidathion
Ito , 1995
113 103
19 14
Pirimiphos-methyl
Ito , 1995
114 104
19 14
Prothiophos
Ito , 1995
115 105
19 14
Pyraclofos
Ito , 1995
116 106
19 14
Tolclofos-methyl
Ito , 1995
117 107
19 14
Trichlorfon
Ito , 1995
118 108
19 14
Vamidothion
Ito , 1995
119 109
19 14
Aspirin
Groten , 1997
91 110
18 15
Cadmium chloride
Groten , 1997
92 111
18 15
Cadmium chloride
Wade , 2002
89 111
17 16
Spermine
Groten , 1997
95 112
18 15
9 181
APPENDIX C
Chemical name
Page 65 of 66
Mode/mechanism, if stated
Mixture study: First
author, Pub. year
CFT/EFSA/PPR/2010/02
ID
Che Mix Art#
m# #
BHA
Groten , 1997
96 113
18 15
dichloromethane
Groten , 1997
98 114
18 15
Formaldehyde
Groten , 1997
99 115
18 15
p,p'-DDE
Wade , 2002
75 117
17 16
p,p'-DDT
Wade , 2002
74 119
17 16
Aldrin
Wade , 2002
73 120
17 16
Dieldrin
Wade , 2002
76 121
17 16
Heptachlor
Wade , 2002
78 122
17 16
Hexachlorobenzene
Wade , 2002
79 123
17 16
Hexachlorocyclohexane
Wade , 2002
80 124
17 16
Methoxychlor
Wade , 2002
82 125
17 16
1,2,4-Trichlorobenzene
Wade , 2002
84 126
17 16
TCDD
Wade , 2002
87 127
17 16
Crofton , 2005
37 127
14 72
Arochlor1254
Wade , 2002
88 128
17 16
Lead chloride
Wade , 2002
90 129
17 16
TCDD
Upregulation of hepatic
catabolism of thyroid hormones
2,4-dinitrophenol
uncoupler of oxidative
phosphorylation
Broderius , 2005
57 139
genistein
not stated
Eustache , 2009
35 153
12 60
162 210
63 177
diclofenac
Rudzok , 2010
6 118
nonylamine
narcotic
Merino-Garcia , 2003
59 223
16 93
decylamine
narcotic
Merino-Garcia , 2003
60 224
16 93
ethyl-parathion
enzyme inhibitor
Merino-Garcia , 2003
61 225
16 93
pentachloroethane
Narcosis I
Broderius , 2005
55 265
4-nitrophenol
Oxidative phosphorylation
uncoupling
Wang , 2009
33 272
2,4-dimethylphenol
Narcosis II
Broderius , 2005
56 274
PCDD
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
38 310
14 72
TCDF
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
39 311
14 72
1-PCDF
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
40 312
14 72
4-PCDF
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
41 313
14 72
OCDF
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
42 314
14 72
PCB-28
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
43 315
14 72
6 118
11 71
6 118
APPENDIX C
Page 66 of 66
CFT/EFSA/PPR/2010/02
Chemical name
Mode/mechanism, if stated
Mixture study: First
author, Pub. year
ID
Che Mix Art#
m# #
PCB-52
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
44 316
14 72
PCB-77
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
45 317
14 72
PCB-101
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
46 318
14 72
PCB-105
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
47 319
14 72
PCB-126
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
49 321
14 72
PCB-138
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
50 322
14 72
PCB-153
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
51 323
14 72
PCB-156
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
52 324
14 72
PCB-169
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
53 325
14 72
PCB-180
Upregulation of hepatic
catabolism of thyroid hormones
Crofton , 2005
54 326
14 72
2,4,6-trichlorophenol
Oxidative phosphorylation
uncoupling
Wang , 2009
34 352
11 71
imidacloprid
Pavlaki , 2011
18 406
8 194
thiacloprid
Pavlaki , 2011
19 407
8 194
nickel
Pavlaki , 2011
20 408
8 194
nickel
Broerse , 2010
21 408
9 181
nickel
Rudzok , 2010
161 408
63 177
chlorobenzene
Nonpolar narcotic
Wang , 2009
25 454
11 71
nitrobenzene
Polar narcotic
Wang , 2009
28 455
11 71
3-nitrotoluene
Polar narcotic
Wang , 2009
29 456
11 71
hydroquinone
Pro-electrophile
Wang , 2009
31 457
11 71
3-nitrochlorobenzene
Pro-electrophile
Wang , 2009
32 458
11 71
Rudzok , 2010
163 486
63 177
Crofton , 2005
48 513
14 72
irgasan
PCB-118
Upregulation of hepatic
catabolism of thyroid hormones