Présentation sur PCB, dioxines, furanes et analyses par DR CALUX (reporter gene) lors du GRMHST du 02.10.2014

1. Groupement romand de médecine,
d’hygiène et de sécurité au travail
Journée de présentation de cas
02 octobre 2014
Exposition aux PCB et Dioxines, qu’en
pense le Dr CALUX ?
Vincent PERRET
Hygiéniste du travail certifié SSHT
Hygiène du travail
Toxicologie industrielle

2. Les PCB quelques points
PolyChloroBiphenyles
209 congénères
2
• Liquides visqueux
• Stables à la chaleur, inertes chimiquement
• Isolant électrique
• Très liposoluble et s’accumulent dans le long de la chaîne alimentaire
• Perturbateurs endoctriniens
• Cancérogènes probables (IARC 2a)

3. Nomenclature des PCB
Nomenclature de Ballschmiter & Zell
3

4. Utilisation des PCB
Usage des PCB répartition
Condensateurs 50.3%
Transformateurs 26.7%
Plastifiants (joints) 9.2%
Huiles hydrauliques 6.4%
Papier carbone 3.6%
Fluides caloporteurs 1.6%
Additifs pétrolier 0.1%
Autres 2.2%
Usage industriel des PCB (1929-1975) – EPA 97
Production mondiale : 1.5 x 106 tonnes (UNEP 98).
4

5. Exemples d’application des
PCB
5

6. Exemples d’application des
PCB
Joints de
séparation
(coupure)
entre
bâtiments
Joints de
raccordement
Joints entre
éléments
Joints de retrait
Joints dans bâtiment (1955-1975)
< 200’000 ppm (20%)
6

7. Exemples d’application des
PCB
7
Peintures industrielles
(1955-1975)
< 20’000 mg/kg (ppm)

8. Exemples d’application des
PCB
Peintures ignifuge de
faux-plafonds
8

9. Exemples d’application des
PCB
Eléments
bitumes
d’étanchéité
de toiture
Source EPA
9

10. Exemples d’application des
PCB
10
Eléments en amiante
ciment
Nombreux cas de
contamination
d’oeufs aux PCBs
dans des fermes du
nord de la Hollande
et d’Allemagne

11. Effets sur la santé
PCB et composés
dioxin-like
11

12. Polychlorodibenzodioxines
(PCDD)
75 congénères
Polychlorodibenzofuranes
(PCDF)
135 congénères
Les PCB
baby dioxine ?
Polychlorobiphényls
(PCB)
209 congénères
12 congénères planaires
12

13. Toxicité relative des
composés dioxines,
furanes et PCB-dl
Facteurs
d’équivalence
toxique relatif au
2,3,7,8- TCDD
13

14. Les grand évènements
impliquant PCBs et dioxines
¡ 1953 Ludwigsfhaven BASF
¡ 1960’ Vietnam, Agent Orange
¡ 1968 Yusho, Japon
¡ 1976 Seveso ICMESA
¡ 2004 Ukraine, Viktor Iouchtchenko
14
Liste non exhaustive, ne manquez pas le prochain épisode

15. SEVESO 15
A : Teq TCDD 15.5 – 580 μg/m2
B : Teq TCDD < 5 μg/m2
C : Teq TCDD < 1.5 μg/m2
Dégâts :
- Fort impact sur végétaux et
animaux (oiseaux)
- Environ 200 cas de chloracné
(88% enfants)
- Modification du sex ratio dans la
région (filles +)
- Augmentation de cancer sujette
à débat

16. Hamster
Golden Syrian
Face à la dioxine,
qui est le plus fort ?
Cochon d’inde
Hartley
16

17. Toxicology Letters 230 (2014) 225–233
4.3. Controversy Although TCDD and the US National of debate regarding A completed report of PCDD TCDD was considered was not conclusive from an application clinical trials in from (1) some et al., 1982; Simon of other 2010; Brookes, between oncogenesis (Seifert Contents lists available at ScienceDirect
Toxicology Letters
journal homepage: www.elsevier.com/locate/toxlet
Mini review
AhR signalling and dioxin toxicity
Olivier Sorg∗
University of Geneva Swiss Centre for Applied Human Toxicology (SCAHT), 1 rue Michel-Servet, 1211 Geneva 4, Switzerland
O. Sorg / Toxicology Letters 230 (2014) 225–233 Table 2
Relation between TCDD LD50 and body fat precentage.
Species (strain) Body fat [%] LD50 [!g/kg]
Guinea pig (Hartley) 4.5 1
American dark mink 4
Hare 7.5 10
Chicken 35
Macaque 10 50
Rat (Sprague-Dawley) 10 50
Dog 100
Rabbit 10 120
Moiuse (C57BL) 8 150
Rat (Fischer) 10 300
Mouse (BALB/c) 400
Dog (Beagle) 14 1000
Frog 1000
Mouse (DBA) 20 2500
Hamster (golden Syrian) 15 10,000
h i g h l i g h t s
• Besides its canonical pathway,
AhR may activate other receptor-mediated
one of the most resistant species with a LD50 of 12.5 mg/kg! How-ever,
pathways.
• AhR activity may be assessed by
chemical or biological assays.
• Dioxin toxicity cannot be explained
only by AhR activation.
• AhR activation leads to either upre-gulation
or downregulation of genes.
• TCDD as a human carcinogen is still a
matter of debate.
• NaturalAhRagonists found in vegeta-bles
might have a beneficial effect.
g r a p h i c a l a b s t r a c t
a r t i c l e i n f o
Article history:
Received 30 August 2013
Received in revised form 14 October 2013
Accepted 18 October 2013
Available online 12 November 2013
Keywords:
Dioxin
TCDD
AhR
Cell signalling
Skin
Toxicity
a b s t r a c t
Dioxins are a family of molecules associated to several industrial accidents such as Ludwigshafen in
1953 or Seveso in 1976, to the Agent Orange used during the war of Vietnam, and more recently to the
poisoning of the former president of Ukraine, Victor Yushchenko. These persistent organic pollutants
are by-products of industrial activity and bind to an intracellular receptor, AhR, with a high potency.
In humans, exposure to dioxins, in particular 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces a
cutaneous syndrome known as chloracne, consisting in the development of many small skin lesions
(hamartoma), lasting for 2–5 years. Although TCDD has been classified by the WHO as a human car-cinogen,
its carcinogenic potential to humans is not clearly demonstrated. It was first believed that AhR
activation accounted for most, if not all, biological properties of dioxins. However, certain AhR agonists
found in vegetables do not induce chloracne, and other chemicals, in particular certain therapeutic agents,
may induce a chloracne-like syndrome without activating AhR. It is time to rethink the mechanism of
dioxin toxicity and analyse in more details the biological events following exposure to these compounds
and other AhR agonists, some of which have a very different chemical structure than TCDD. In particular
various food-containing AhR agonists are non-toxic and may on the contrary have beneficial properties
to human health.
© 2013 Elsevier Ireland Ltd. All rights reserved.
Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
2. AhR signalling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
17

18. Effets aigüs
Le cas Iouchtchenko
2000 2004 Empoisonnement
au TCDD de Viktor
Iouchtchenko (alors
premier ministre
ukrainien)
(6 sept 2004)
18

19. TOXICOLOGICAL SCIENCES 125(1), 310–317 (2012)
doi:10.1093/toxsci/kfr223
Advance Access publication October 13, 2011
19
The Cutaneous Lesions of Dioxin Exposure: Lessons from the Poisoning
of Victor Yushchenko
Jean-Hilaire Saurat,*,†,1 Guerkan Kaya,*,† Nikolina Saxer-Sekulic,*,† Bruno Pardo,* Minerva Becker,‡ Lionel Fontao,†
Florence Mottu,*,† Pierre Carraux,† Xuan-Cuong Pham,† Caroline Barde,† Fabienne Fontao,* Markus Zennegg,§
Peter Schmid,§ Olivier Schaad,{ Patrick Descombes,{ and Olivier Sorg*,†
*Swiss Centre for Applied Human Toxicology, Dermatotoxicology Unit, University of Geneva, 1211 Geneva 4, Switzerland; †Dermatology Department and
‡Radiology Department, Geneva University Hospital, 1211 Geneva 14, Switzerland; §EMPA (Swiss Federal Laboratories for Materials Testing and Research),
8600 Du¨bendorf, Switzerland; and {Genomics Platform, National Center of Competence in Research Frontiers in Genetics, University of Geneva, 1211
Geneva 4, Switzerland
1To whom correspondence should be addressed at Swiss Centre for Applied Human Toxicology, University of Geneva, 1, rue Michel-Servet, 1211 Gene`ve 4,
Switzerland. Fax: 0041-22-379 5502. E-mail: jean.saurat@unige.ch.
Received August 10, 2011; accepted August 10, 2011
Several million people are exposed to dioxin and dioxin-like
characterization of dioxin exposure remains difficult to establish,
… pour la Science

20. of sampling were prepared as previously reported (Sorg et al., 2008). RNA
TOXICOLOGICAL SCIENCES 125(1), 310–317 (2012)
quality doi:10.1093/toxsci/was kfr223
assessed using an Agilent 2100 Bioanalyzer with an RNA 6000
Advance Access publication October 13, 2011
Nano LabChip kit. We generated a hybridization mixture containing 15 lg of
biotinylated complementary RNA and hybridized it to GeneChip HG U133
Plus 2.0 according to manufacturer’s instructions (Affymetrix). To identify
differentially expressed transcripts, comparisons were carried out after
normalization with the Affymetrix GCOS 1.2 (MAS5) software.
The Cutaneous Lesions of Dioxin Exposure: Lessons from the Poisoning
of Victor Yushchenko
Jean-Hilaire Saurat,*,†,1 Guerkan Kaya,*,† Nikolina Saxer-Sekulic,*,† Bruno Pardo,* Minerva Becker,‡ Lionel Fontao,†
Florence Mottu,*,† Pierre Carraux,† Xuan-Cuong Pham,† Caroline Barde,† Fabienne Fontao,* Markus Zennegg,§
Bioinformatic analysis. Responsive elements for genes corresponding to
Peter Schmid,§ Olivier Schaad,{ Patrick Descombes,{ and Olivier Sorg*,†
differentially expressed transcripts were searched with the University of
small fiber peripheral neuropathy (The organs and systems
involved, other than the skin, are just cited here. Each will be
fully addressed in future publications when the mechanism has
been better analyzed by appropriate ongoing data analysis.).
Facial involvement worsened with diffuse nodular lesions on an
edematous background, sparing of the periocular zone, but
major involvement of the ears and retroauricular folds (Figs. 2A
and 2B). The basic skin lesions were small nodules (Fig. 2C),
*Swiss Centre for Applied Human Toxicology, Dermatotoxicology Unit, University of Geneva, 1211 Geneva 4, Switzerland; †Dermatology Department and
‡Radiology Department, Geneva University Hospital, 1211 Geneva 14, Switzerland; §EMPA (Swiss Federal Laboratories for Materials Testing and Research),
8600 Du¨bendorf, Switzerland; and {Genomics Platform, National Center of Competence in Research Frontiers in Genetics, University of Geneva, 1211
Geneva 4, Switzerland
1To whom correspondence should be addressed at Swiss Centre for Applied Human Toxicology, University of Geneva, 1, rue Michel-Servet, 1211 Gene`ve 4,
Switzerland. Fax: 0041-22-379 5502. E-mail: jean.saurat@unige.ch.
Received August 10, 2011; accepted August 10, 2011
Several million people are exposed to dioxin and dioxin-like
compounds, primarily through food consumption. Skin lesions
historically called ‘‘chloracne’’ are the most specific sign of
abnormal dioxin exposure and classically used as a key marker in
humans. We followed for 5 years a man who had been exposed to
the most toxic dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin
(TCDD), at a single oral dose of 5 million-fold more than the
accepted daily exposure in the general population. We adopted
a molecular medicine approach, aimed at identifying appropriate
therapy. Skin lesions, which progressively covered up to 40% of
the body surface, were found to be hamartomas, which developed
parallel to a complete and sustained involution of sebaceous
glands, with concurrent transcriptomic alterations pointing to the
inhibition of lipid metabolism and the involvement of bone
morphogenetic proteins signaling. Hamartomas created a new
compartment that concentrated TCDD up to 10-fold compared
with serum and strongly expressed the TCDD-metabolizing
enzyme cytochrome P450 1A1, thus representing a potentially
significant source of enzymatic activity, which may add to the
xenobiotic metabolism potential of the classical organs such as the
liver. This historical case provides a unique set of data on the
human tissue response to dioxin for the identification of new
markers of exposure in human populations. The herein discovered
adaptive cutaneous response to TCDD also points to the potential
role of the skin in the metabolism of food xenobiotics.
Key Words: dioxin; toxicity; skin; hamartoma; morphology.
Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD) is the
most potent of a large number of industrial-era halogenated
polyaromatic hydrocarbon pollutants, including other dibenzo-p-dioxins,
dibenzofurans, and certain polychlorinated biphenyls.
Human populations are exposed to low levels of dioxin and
dioxin-like compounds, primarily through food consumption
(Connor et al., 2008; Schecter et al., 1999). The risk
characterization of dioxin exposure remains difficult to establish,
although it is an issue that broadly affects important public health
policy decisions (Gies et al., 2007; Steenland et al., 2001). Thus,
chronic exposure to low/moderate doses of dioxin may be
involved not only in the classic dioxin toxicity in some
genetically predisposed individuals (IARC Monograph, 1997;
Aylward et al., 2005) but also in the newly identified role of
these compounds in autoimmunity (Brembilla et al., 2011;
Marshall and Kerkvliet, 2010; Ramirez et al., 2010).
In humans, skin lesions called ‘‘chloracne’’ are the most
visible and consistent response to dioxin exposure and therefore
play the role of a sentinel sign for toxicity (Caputo et al., 1988).
The mechanism by which chloracne appears was not previously
known and its diagnostic value is not straightforward, especially
in mild and sporadic cases, which could still relate to significant
exposure (Passarini et al., 2010). A robust indicator that would
trigger specific ecotoxicology diagnostic processes is lacking; in
the current situation, it is likely that many cases have not been
recognized (Saurat and Sorg, 2010).
We have previously reported on the TCDD poisoning in
Victor Yushchenko with identification and measurement of
TCDD metabolites (Sorg et al., 2009). The maximum accepted
daily dose exposure in human is 4 pg/kg, and this patient
received a single dose of 20 lg/kg.
With the approval of the patient to release peer-reviewed
scientific information on his case, we now report on a set of
data that has never been obtained in humans and helps define
the phenotype of the dioxin-induced skin pathology.
FIG. 1. Evolution of the dioxin disease. Chronology of organ involvement a.p. The peak of skin involvement is delayed as compared with the other organs,
and skin lesions show a longer and chronic course.
MATERIALS AND METHODS
Clinical specimens. Skin sampling was performed under general anesthe-sia
during therapeutic procedures.
! The Author 2011. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved.
For permissions, please email: journals.permissions@oup.com
Downloaded from http://toxsci.oxfordjournals.org/ by guest on October 1, 2014
20

21. SCIENCES 125(1), 310–317 (2012)
Downloaded TOXICOLOGICAL doi:10.1093/toxsci/kfr223
Advance Access publication October 13, 2011
The Cutaneous Lesions of Dioxin Exposure: Lessons from the Poisoning
FROM THE POISONING OF VICTOR YUSHCHENKO 315
sebum lipid
! Downloaded the promoter
and sterol O-acyltransferase
tissue renewing
induced genes,
factor A serine
of the bone
highly relevant to
al., 2006).
from structural/
extracellular matrix’’
expected from
specific pattern.
strongly repressed
involved in the
of Victor Yushchenko
Jean-Hilaire Saurat,*,†,1 Guerkan Kaya,*,† Nikolina Saxer-Sekulic,*,† Bruno Pardo,* Minerva Becker,‡ Lionel Fontao,†
Florence Mottu,*,† Pierre Carraux,† Xuan-Cuong Pham,† Caroline Barde,† Fabienne Fontao,* Markus Zennegg,§
Peter Schmid,§ Olivier Schaad,{ Patrick Descombes,{ and Olivier Sorg*,†
*Swiss Centre for Applied Human Toxicology, Dermatotoxicology Unit, University of Geneva, 1211 Geneva 4, Switzerland; †Dermatology Department and
‡Radiology Department, Geneva University Hospital, 1211 Geneva 14, Switzerland; §EMPA (Swiss Federal Laboratories for Materials Testing and Research),
8600 Du¨bendorf, Switzerland; and {Genomics Platform, National Center of Competence in Research Frontiers in Genetics, University of Geneva, 1211
Geneva 4, Switzerland
1To whom correspondence should be addressed at Swiss Centre for Applied Human Toxicology, University of Geneva, 1, rue Michel-Servet, 1211 Gene`ve 4,
Switzerland. Fax: 0041-22-379 5502. E-mail: jean.saurat@unige.ch.
Received August 10, 2011; accepted August 10, 2011
Several million people are exposed to dioxin and dioxin-like
compounds, primarily through food consumption. Skin lesions
historically called ‘‘chloracne’’ are the most specific sign of
abnormal dioxin exposure and classically used as a key marker in
humans. We followed for 5 years a man who had been exposed to
the most toxic dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin
(TCDD), at a single oral dose of 5 million-fold more than the
accepted daily exposure in the general population. We adopted
a molecular medicine approach, aimed at identifying appropriate
therapy. Skin lesions, which progressively covered up to 40% of
the body surface, were found to be hamartomas, which developed
parallel to a complete and sustained involution of sebaceous
glands, with concurrent transcriptomic alterations pointing to the
inhibition of lipid metabolism and the involvement of bone
morphogenetic proteins signaling. Hamartomas created a new
compartment that concentrated TCDD up to 10-fold compared
with serum and strongly expressed the TCDD-metabolizing
enzyme cytochrome P450 1A1, thus representing a potentially
significant source of enzymatic activity, which may add to the
xenobiotic metabolism potential of the classical organs such as the
liver. This historical case provides a unique set of data on the
human tissue response to dioxin for the identification of new
markers of exposure in human populations. The herein discovered
adaptive cutaneous response to TCDD also points to the potential
role of the skin in the metabolism of food xenobiotics.
Key Words: dioxin; toxicity; skin; hamartoma; morphology.
Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD) is the
most potent of a large number of industrial-era halogenated
polyaromatic hydrocarbon pollutants, including other dibenzo-p-dioxins,
dibenzofurans, and certain polychlorinated biphenyls.
Human populations are exposed to low levels of dioxin and
dioxin-like compounds, primarily through food consumption
(Connor et al., 2008; Schecter et al., 1999). The risk
characterization of dioxin exposure remains difficult to establish,
although it is an issue that broadly affects important public health
policy decisions (Gies et al., 2007; Steenland et al., 2001). Thus,
chronic exposure to low/moderate doses of dioxin may be
involved not only in the classic dioxin toxicity in some
genetically predisposed individuals (IARC Monograph, 1997;
Aylward et al., 2005) but also in the newly identified role of
these compounds in autoimmunity (Brembilla et al., 2011;
Marshall and Kerkvliet, 2010; Ramirez et al., 2010).
In humans, skin lesions called ‘‘chloracne’’ are the most
visible and consistent response to dioxin exposure and therefore
play the role of a sentinel sign for toxicity (Caputo et al., 1988).
The mechanism by which chloracne appears was not previously
known and its diagnostic value is not straightforward, especially
in mild and sporadic cases, which could still relate to significant
exposure (Passarini et al., 2010). A robust indicator that would
trigger specific ecotoxicology diagnostic processes is lacking; in
the current situation, it is likely that many cases have not been
recognized (Saurat and Sorg, 2010).
We have previously reported on the TCDD poisoning in
Victor Yushchenko with identification and measurement of
TCDD metabolites (Sorg et al., 2009). The maximum accepted
daily dose exposure in human is 4 pg/kg, and this patient
received a single dose of 20 lg/kg.
With the approval of the patient to release peer-reviewed
scientific information on his case, we now report on a set of
data that has never been obtained in humans and helps define
the phenotype of the dioxin-induced skin pathology.
MATERIALS AND METHODS
Clinical specimens. Skin sampling was performed under general anesthe-sia
during therapeutic procedures.
from http://toxsci.oxfordjournals.org/ by guest on October 1, 2014
21

22. Compassionate use of tumor necrosis factor a (TNF-a) blockade
was considered because non-steroidal anti-inflammatory drugs and
systemic steroids were not effective. The patient received three
infusions of infliximab but because of intolerancewas then switched
to adalimumab, which was given for M18 (M16th to M34th a.p.).
AL.
! Downloaded FIG. 5. Volumetric analyses of the skin lesions. (A) Three-dimensional
representation of the calculated total skin volume of the face using the
methodology described in the text. (B) Three-dimensional representation of the
calculated volume of the hamartomatous lesions seen in the same area as in (A).
Anterior view. Note that most lesions are located in the ear lobes and lateral
cheeks. (C) Thick slab reconstruction of an FDG PET data set obtained from
a whole-body PET/CT acquisition showing the distribution and the metabolism
org/ by guest on October 1, 2014
TOXICOLOGICAL SCIENCES 125(1), 310–317 (2012)
doi:10.1093/toxsci/kfr223
Advance Access publication October 13, 2011
The Cutaneous Lesions of Dioxin Exposure: Lessons from the Poisoning
of Victor Yushchenko
Jean-Hilaire Saurat,*,†,1 Guerkan Kaya,*,† Nikolina Saxer-Sekulic,*,† Bruno Pardo,* Minerva Becker,‡ Lionel Fontao,†
Florence Mottu,*,† Pierre Carraux,† Xuan-Cuong Pham,† Caroline Barde,† Fabienne Fontao,* Markus Zennegg,§
Peter Schmid,§ Olivier Schaad,{ Patrick Descombes,{ and Olivier Sorg*,†
*Swiss Centre for Applied Human Toxicology, Dermatotoxicology Unit, University of Geneva, 1211 Geneva 4, Switzerland; †Dermatology Department and
‡Radiology Department, Geneva University Hospital, 1211 Geneva 14, Switzerland; §EMPA (Swiss Federal Laboratories for Materials Testing and Research),
8600 Du¨bendorf, Switzerland; and {Genomics Platform, National Center of Competence in Research Frontiers in Genetics, University of Geneva, 1211
Geneva 4, Switzerland
1To whom correspondence should be addressed at Swiss Centre for Applied Human Toxicology, University of Geneva, 1, rue Michel-Servet, 1211 Gene`ve 4,
Switzerland. Fax: 0041-22-379 5502. E-mail: jean.saurat@unige.ch.
Received August 10, 2011; accepted August 10, 2011
Several million people are exposed to dioxin and dioxin-like
compounds, primarily through food consumption. Skin lesions
historically called ‘‘chloracne’’ are the most specific sign of
abnormal dioxin exposure and classically used as a key marker in
humans. We followed for 5 years a man who had been exposed to
the most toxic dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin
(TCDD), at a single oral dose of 5 million-fold more than the
accepted daily exposure in the general population. We adopted
a molecular medicine approach, aimed at identifying appropriate
therapy. Skin lesions, which progressively covered up to 40% of
the body surface, were found to be hamartomas, which developed
parallel to a complete and sustained involution of sebaceous
glands, with concurrent transcriptomic alterations pointing to the
inhibition of lipid metabolism and the involvement of bone
morphogenetic proteins signaling. Hamartomas created a new
compartment that concentrated TCDD up to 10-fold compared
with serum and strongly expressed the TCDD-metabolizing
enzyme cytochrome P450 1A1, thus representing a potentially
significant source of enzymatic activity, which may add to the
xenobiotic metabolism potential of the classical organs such as the
liver. This historical case provides a unique set of data on the
human tissue response to dioxin for the identification of new
markers of exposure in human populations. The herein discovered
adaptive cutaneous response to TCDD also points to the potential
role of the skin in the metabolism of food xenobiotics.
Key Words: dioxin; toxicity; skin; hamartoma; morphology.
Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD) is the
most potent of a large number of industrial-era halogenated
polyaromatic hydrocarbon pollutants, including other dibenzo-p-dioxins,
dibenzofurans, and certain polychlorinated biphenyls.
Human populations are exposed to low levels of dioxin and
dioxin-like compounds, primarily through food consumption
(Connor et al., 2008; Schecter et al., 1999). The risk
characterization of dioxin exposure remains difficult to establish,
although it is an issue that broadly affects important public health
policy decisions (Gies et al., 2007; Steenland et al., 2001). Thus,
chronic exposure to low/moderate doses of dioxin may be
involved not only in the classic dioxin toxicity in some
genetically predisposed individuals (IARC Monograph, 1997;
Aylward et al., 2005) but also in the newly identified role of
these compounds in autoimmunity (Brembilla et al., 2011;
Marshall and Kerkvliet, 2010; Ramirez et al., 2010).
In humans, skin lesions called ‘‘chloracne’’ are the most
visible and consistent response to dioxin exposure and therefore
play the role of a sentinel sign for toxicity (Caputo et al., 1988).
The mechanism by which chloracne appears was not previously
known and its diagnostic value is not straightforward, especially
in mild and sporadic cases, which could still relate to significant
exposure (Passarini et al., 2010). A robust indicator that would
trigger specific ecotoxicology diagnostic processes is lacking; in
the current situation, it is likely that many cases have not been
recognized (Saurat and Sorg, 2010).
We have previously reported on the TCDD poisoning in
Victor Yushchenko with identification and measurement of
TCDD metabolites (Sorg et al., 2009). The maximum accepted
daily dose exposure in human is 4 pg/kg, and this patient
received a single dose of 20 lg/kg.
With the approval of the patient to release peer-reviewed
scientific information on his case, we now report on a set of
data that has never been obtained in humans and helps define
the phenotype of the dioxin-induced skin pathology.
MATERIALS AND METHODS
Clinical specimens. Skin sampling was performed under general anesthe-sia
during therapeutic procedures.
from http://toxsci.oxfordjournals.org/ by guest on October 1, 2014
22
FIG. 3. Histological analyses of the skin lesions. (A) Photomicrograph of

23. Prélèvements et
analyses
PCB et composés
dioxin-like
23

24. 24
Ordonnance sur les travaux de
construction
Obligation de planification des
travaux si la présence de PCB est
suspectée

25. fixe une limite de teneur
de PCB dans les joints
de
50 ppm (mg/kg)
Appliqué par extension
aux autres matériaux
(peintures)
25

26. 26

27. Analyse des PCBs
27

28. Les PCB indicateurs
les 6 grands électeurs
PCB 28 PCB 101 PCB 153
PCB 52 PCB 138 PCB 180
28

29. 1016$
1242$
1248$
1248$
1254$"Late"$
1254$
1260$
0$
2$
4$
6$
8$
10$
12$
14$
1$
4$
7$
10$
13$
16$
19$
22$
25$
28$
31$
34$
37$
40$
43$
46$
49$
52$
55$
58$
61$
64$
67$
70$
73$
76$
79$
82$
85$
88$
91$
94$
97$
100$
103$
106$
109$
112$
115$
118$
121$
124$
127$
130$
133$
136$
139$
142$
145$
148$
151$
154$
157$
160$
163$
166$
169$
172$
175$
178$
181$
184$
187$
190$
193$
196$
199$
202$
205$
208$
%"masse"
1016$ 1242$ 1248$ 1248$ 1254$"Late"$ 1254$ 1260$
La composition des Aroclors
29
Données sources : ATDSR

30. Les facteurs de conversion 30
version massique
28 52 101 138 153 180 Total % Facteur
1016 8.50 4.63 0.04 13.2 7.6
1242 6.86 3.53 0.69 0.10 0.06 11.2 8.9
1248 3.59 6.93 2.22 0.38 0.23 0.02 13.4 7.5
1248 5.57 5.58 1.89 0.41 0.43 0.21 14.1 7.1
1254 "Late" 0.06 0.83 5.49 5.95 3.29 0.42 16.0 6.2
1254 0.19 5.38 8.02 5.80 3.77 0.67 23.8 4.2
1260 0.03 0.24 3.13 6.54 9.39 11.38 30.7 3.3
Aroclor
PCB

31. Et au niveau toxicité des 31
PCB-dl ?
77 0.0001 81 0.0003 126 0.1 169 0.03 105 0.00003 114 0.00003 118 0.00003 123 0.00003 156 0.00003 157 0.00003 167 0.00003 189 0.00003
%masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq %masse Teq
1016
1242 0.31 3E-05 0.01 3E-06 0.47 1E-05 0.04 1E-06 0.66 2E-05 0.03 9E-07 0.01 3E-07
1248 0.41 4E-05 0.01 3E-06 1.6 5E-05 0.12 4E-06 2.29 7E-05 0.07 2E-06 0.06 2E-06 0.01 3E-07 0.01 3E-07
1248 0.52 5E-05 0.02 6E-06 1.45 4E-05 0.12 4E-06 2.35 7E-05 0.08 2E-06 0.04 1E-06 0.01 3E-07
1254 "Late" 0.2 2E-05 0.02 0.002 7.37 0.0002 0.5 2E-05 13.59 4E-04 0.32 1E-05 1.13 3E-05 0.3 9E-06 0.35 1E-05 0.01 3E-07
1254 0.03 3E-06 2.99 9E-05 0.18 5E-06 7.35 2E-04 0.15 5E-06 0.82 2E-05 0.19 6E-06 0.27 8E-06 0.01 3E-07
1260 0.22 7E-06 0.48 1E-05 0.52 2E-05 0.02 6E-07 0.19 6E-06 0.1 3E-06
Teq (ng/g) pour 50 ppm
28 52 101 138 153 180 Total % Facteur
1016 8.50 4.63 0.04 13.2 7.6
1242 6.86 3.53 0.69 0.10 0.06 11.2 8.9
1248 3.59 6.93 2.22 0.38 0.23 0.02 13.4 7.5
1248 5.57 5.58 1.89 0.41 0.43 0.21 14.1 7.1
1254 "Late" 0.06 0.83 5.49 5.95 3.29 0.42 16.0 6.2
1254 0.19 5.38 8.02 5.80 3.77 0.67 23.8 4.2
1260 0.03 0.24 3.13 6.54 9.39 11.38 30.7 3.3
Aroclor
PCB
Teq$ng/kg
1016 0
1242 5
1248 39
1248 41
1254 "Late" 3244
1254 217
1260 4

32. OK, les PCB
indicateurs c’est
pourri,
mais comment
on peut mesurer
un Teq TCDD ? La question que le
premier rang se pose

33. GC-HRMS 33
Profil GC-HRMS d’un extrait de
cendres volantes. © Restek

34. pg eq TCDD /
gr échantillon
34
Oui, mais
7 PCDD sur 75
10 PCDF sur 135
12 PBC-dl sur 12
… et les autres ligands ?
… et les synergies ?

35. Au secours
DR CALUX !!!
Dr Heinrich von Calux
1875 - 1910
Peut-on mesure35r
réellement un
Teq TCDD alors ?

36. Dioxin
Receptor
Chemically
Activated
LUciferase gene
eXpression
36
Vous allez
voir, c’est
tout simple
Méthode de biologie
moléculaire de mesure
de l’induction d’un
récepteur par un ligand.

37. 37
Voyez,
c’est simple

38. 38
NOYAU
Moins
simple

39. 39
… désolé

40. Principe du gène rapporteur
(reporteur gene)
40

41. DR-CALUX méthodologie
41

42. GC-HRMS vs DR-CALUX
42
GC-HRMS DR-CALUX
Spécificité +++
Substances
+
Capacité d’activation du
récepteur
Sensibilité ++
1 pg eq TCDD
+++
0.3 pg eq TCDD
Capacité de screening ++
Analyse par substance
+++
Détection des ligands
inconnus
Détection des effets de
synergie (coktail)
Mise en oeuvre Coût : env 1000 CHF
Rapidité : env 15 jours
Coût : env 250 CHF
Rapidité : env 5 jours
Validation Golden Standard Validations, food, sang, terres,
poussières, eaux.

43. Exemples d’application du DR CALUX en
hygiène du travail
Suivi des expositions des travailleurs lors des travaux de
sécurisation d’un site contaminé aux PCB.
43

44. Mesure de la perméation des PCB
indicateurs dans les combinaisons

45. Suivi sanguin DR-CALUX
analyse Teq TCDD sang
A (avant travaux):
21.8 +/- 5.03 pg TCDD TEQ/g fat
B: (après travaux):
19.9 +/- 6.42 pg TCDD TEQ/g fat
Pas de différence statistiquement significative
entre les 2 séries (Wilcoxon signed rank test,
paired values)
No patient
1
2
3
4
5
6
7
8
9
10
Average
Median
Standard deviation
Coeff. of variation
Minimum
Maximum
Range
Stnd. skewness
Stnd. kurtosis
CALUX A
PCDD/PCDF and dl-
PCBs (only total TEQ)
[pg TEQ/g fat]
CALUX B
PCDD/PCDF and dl-
PCBs (only total TEQ)
[pg TEQ/g fat]
19* 29
28 25
32 18
20 16
24 16
20 13
16* 17
23 21
19 31
17 13
21.8 19.9
20 17.5
5.03 6.42
0.23 0.32
16 13
32 31
16 18
1.34 0.98
0.31 -0.50
(*) < LOQ
Concentrations usuelles dans le sérum
(publications)
Non exposé ou nouveau né:
23 to 27 pg TCDD TEQ/g fat
Habitant proche d’une usine d’incinération
55 to 76 pg TCDD TEQ/g fat
T0 T+4 mois
45

46. Exemples d’application du DR CALUX en
environnement
Analyse des PCB dans des éléments en amiante-ciment
46

47. Concentration*TCDD-TEQ
Etat pg/g*TEQ pg/cm2*TEQ
Site*A*-*gris*clair <*0.067 <*0.12
Site*A*-*gris*foncé*après*incendie 1.900 0.50
Site*B*-*peinture*de*surface*rouge 0.940 2.70
Site*C*-*gris*clair 0.400 0.34
Site*D*-*gris*foncé 0.066 0.07
Site*E*-*gris*foncé 0.037 0.05
Site*F*-*gris*clair 0.029 0.03
Site*G*-*gris*clair 0.061 0.05
47
Ech.%2 Ech.%3
PCB$indicateurs pg/g pg/g
PCB'28 <5.8 <0.24
PCB'52 <5.8 14
PCB'101 28 58
PCB'138 103 57
PCB'153 71 46
PCB'180 141 13
Somme'des'7'PCB'indicateurs 344 188
1.72 ppm < 50 ppm
Ech.%2 Ech.%3
PCB$dioxin$like pg/g pg/g
PCB'77 <5.8 11
PCB'105 81 29
PCB'114 <5.8 <0.24
PCB'118 49 30
PCB'123 <5.8 <0.24
PCB'126 <5.8 1.6
PCB'156 <5.8 8.3
PCB'157 <5.8 2.6
PCB'167 <5.8 3.9
PCB'169 <5.8 <0.24
PCB'189 <5.8 <0.24
WHO(2005)'PCB'TEQ 2.5 0.19
8 échantillons d’éléments en amiante-ciment
(Suisse Romande)
TOXpro 2014, unpublished data

48. Toiture :
13 pg/g PCB-dl (Teq TCDD)
Prélèvement de surface
345’141 pg/g PCB-dl (Teq TCDD)
Sol
139 pg/g PCD-dl (Teq TCDD)
48

49. Exemples d’application du DR CALUX en
hygiène du travail
Contamination des surfaces de travail aux composés Dioxin-like
dans un incinérateur municipal

50. 50
Prélèvement des surfaces (wipe-test)
TOXpro 2014, unpublished data
Valeurs seuils
Allemagne
10 ng/m2

51. SIG$D13$($fines$sous$grille$
2378*TetraCDD#
40%#
30%#
20%#
10%#
0%#
12378*PentaCDD#
123478*HexaCDD#
123678*HexaCDD#
123789*HexaCDD#
1234678*HeptaCDD#
12346789*OctaCDD#
2378*TetraCDF#
12378*PentaCDF#
23478*PentaCDF#
123478*HexaCDF#
123678*HexaCDF#
123789*HexaCDF#
234678*HexaCDF#
12346789*OctaCDF#
1234789*HeptaCDF# 1234678*HeptaCDF#
PCB#77#
PCB#81#
PCB#123#
PCB#118#
PCB#114#
PCB#169#
PCB#126#
PCB#105#
PCB#156#
PCB#157#
PCB#167#
PCB#189#
PCDD$
PCDF$
PCB'dl$
SIG$D3$'$Cendres$volantes$
2378*TetraCDD#
40%#
30%#
20%#
10%#
0%#
12378*PentaCDD#
123478*HexaCDD#
123678*HexaCDD#
123789*HexaCDD#
1234678*HeptaCDD#
12346789*OctaCDD#
2378*TetraCDF#
12378*PentaCDF#
23478*PentaCDF#
123478*HexaCDF#
123678*HexaCDF#
123789*HexaCDF#
234678*HexaCDF#
12346789*OctaCDF#
1234789*HeptaCDF# 1234678*HeptaCDF#
PCB#77#
PCB#81#
PCB#123#
PCB#118#
PCB#114#
PCB#169#
PCB#126#
PCB#105#
PCB#156#
PCB#157#
PCB#167#
PCB#189#
PCDD$
PCDF$
PCB'dl$
51
Recherche des sources par GC-HRMS
Cendres volantes Dépôt sur armoire
TOXpro 2014, unpublished data

52. Mais bon,
y’a pas que les
PCB / Dioxines
dans la vie !
PCB et composés
dioxin-like

53. Les retardateurs de flamme
Les retardateurs de flamme
53

54. 54
Brommer et al. J
Environ Monitor
2012;
Marklund et al.
Chemosphere 2003;
Stapleton et al.
Environ Sci Technol
2005;2009;
Takigami et al.
Environ Int 2009; Van
den Eede et al.
Environ Int 2011

55. profiles to in vivo toxicity of the compounds?
La gamme des récepteurs et « pathway) 55
compound ‘profile’
Compound
CALUX assays currently available:
Nuclear receptors Signaling pathways
name endpoint name endpoint
DR CALUX dioxins NFκB CALUX inflammation
PAH CALUX PAHs p21 CALUX DNA damage
ERα CALUX estrogens Nrf2 CALUX oxid. stress
ERβ CALUX estrogens p53 CALUX DNA damage
AR CALUX androgens TCF CALUX carcinogenesis
PR CALUX progestins AP1 CALUX stress
GR CALUX glucocortocoid HIF1α CALUX hypoxia
TRβ CALUX thyroids ESRE CALUX ER stress
RAR CALUX retinoids Cytotox CALUX cytotoxicity
PPARγ CALUX obesogens
PPARα CALUX obesogens
PPARδ CALUX obesogens
PXR CALUX xenobiotics
LXR CALUX oxysterols

56. Les possibilités de screening 56

57. Screening des
micropolluants, eaux uséés
57

58. OUF !
58