Conferencia Dr. Carrasco

1. “This time results of an
unlimited license granted to
technology” and....
“constitute the perfect crime
to kill reality”
Jean Baudrillard
Andres E. Carrasco
Laboratorio Embriologia Molecular
Facultad de Medicina UBA-CONICET

2. Argentina and agrotoxics
and
Territorial conflicts

4. El
futuro:
P.A.N

5. Soy in Argentina
Extension: 20.000.000 Ha.
GBH: 200.000.000 liters year , (Paraguay GBH 8 liters year)
Population living in the territory 10.000.000 people.
70 % of the effects are by inhalation.

6. Increasing of use of plaguicides 1999-2009

7. Preliminary epidemiological humans
studies in Argentina

8. Birth prevalence of 27 selected congenital anomalies in 7 geographic
regions of Argentina ,
Campaña et al (ECLAMC group). Arch Argent. Pediatr. 108 (5) 409-417, 2010
Out of the 27 congenital anomalies analyzed, 14 showed a frequency
significatively higher in one or more regions
Centro spina bifida, microtia, cleft lip with cleft palate, polycystic kidney,
postaxial polydactyly and Down Syndrome,
Northest omphalocele, gastroschisis, cleft lip with and without cleft palate,
anorectal atresia/stenosis, indeterminate sex, preaxial polydatily and pectoral
agenesis,
Patagonia cleft lip with cleft palate.
Cuyo postaxial polydactyly,
Pampa, severe hypospadias

9. CORDOBA

10. Obstetric and Neonatology Hospital, Cordoba Univ, (Dra. Trombotto)
On 111.000 newborns,
CRANEOFACIAL , GASTROSCHISIS, LIMB DEFORMITIES
Incidence of MCM (not due to genetics or familiar bias)
1991: 16.2 %o newborns.
2003 : 37.1 %o newborns.

11. Ituzango, Cordoba.
RED DOTS: CANCER CASES

12. Chaco, Argentina
Departamento
Bermejo
Puerto Eva Perón
General
Vedia
Puerto Bermejo
LA LEONESA
Las Palmas
Isla del Cerrito

13. La Leonesa, Dept. Rio Bermejo, Chaco
Población Menores
Casos Esperados Casos Incidencia
Municipio Total de 15 años
Ca Infantil - por año Registrados años
2001 - 2001
1995: 1 caso 1990-1999
1997:
0,41 casos por año 2000:
1 “
1 ”
0,2 casos/año –
1 caso/60meses
300%
La
10.067 2960 ( 1 caso c/ 24- 36 2003: 2 “
Leonesa 2000-2009
meses) 2004: 1 “
2008: 1 “ 0,6 casos/año – 1 caso/20
2009: 1 “ meses
1990-1999
0,2 casos/año
0,3 casos por año 1993: 1 “ 1 caso/60 meses
Las
6593 2146 ( 1 caso c/ 36-42 1995: 1 “
Palmas 2000-2009
meses) 2006: 1 “
0,1 caso/año
1 caso/ 120 meses
1990-1999
0,1 caso/año
0,09 casos x año
Puerto 1995: 1 caso 1 caso/20 meses
1832 652 ( 1 caso c/ 96
Bermejo 2008: 1 “ 2000-2009
años)
0,1 caso/año
1 caso/120 meses

14. Malformations % in total live newborns
Año N /year Live born Incidency
(malformations *10000/
live born )
1997 46 malformacions 24030 (live born 1997) 19,1 /10.000
2001 60 malformacions 21339 (live born 2001) 28,1 /10.000
2008 186 malformacions 21.808 (live born 2008) 85,3 /10.000
400%
en
los
ul,mos
10
años

15. Benitez Leite et al, Rev Ped. Paraguay (2009)
139 newborns were analyzed.
Where founded 52 malformations
associated to exposure to agrotoxics
(plus 34 death fetus).
In Roque Saenz Peña, Chaco, Argentine.
In 4 years, over 4255 of newborns, 157
malformations were detected (average 3.6%)

16. Documents:
CONICET (2009)
UNIV. DEL LITORAL (2010)
GM SOY: SUSTAINABLE?
RESPONSIBLE? (2010)
ROUNDUP and BIRTH DEFECTS
(2011)

17. PESTICIDES USED IN SOUTH AMERICAN
GMO-BASED AGRICULTURE: A REVIEW OF
THEIR EFFECTS ON HUMANS AND ANIMAL
MODELS
Silvia L. López; Delia Aiassa; Stella Benítez-Leite; Rafael
Lajmanovich; Fernando Mañas; Gisela Poletta; Norma
Sánchez; María Fernanda Simoniello; and Andrés E.
Carrasco.
Advances in Molecular Toxicology
Vol 6, 2012, (in press)

18. Legal actions to block effects of agrochemicals
San Jorge, Santa Fe.
Sentence: 800 m. for ground machine and 1500 m. aerial aspersion.
Legal action. Santa Fe´s Court of Apppels.
La Leonesa, Chaco.
Sentence: 1000 m. for ground machine and 2000 m. aerial aspersion.
Legal action. Chaco’s Superior Court .
National Ombusman.
Resolution recommending to the Minister of Agriculture to review
classification of all agrochemicals used in Argentina.

19. Glyphosate and teratogenesis

20. Experimental evidences of TOXICITY of
AGROCHEMICAL
GLYPHOSATE
 LOCK THE DNA REPAIR MACHINERY. (checkpoint enzymes)
B
 LOCK CELL CYCLE DURING DEVELOMENT.
B
 CTIVATE APOPTOSIS IN CELL CULTURE.
A
 RODUCE TERATOGENESIS DURING VERTEBRATE
P
DEVELOMENT (AVIAN, ANPHIBIAN, MAMMALIANS)

21. Laboratorio de Embriología Molecular
Facultad de Medicina
Universidad de Buenos Aires – CONICET
acarrasco@fmed.uba.ar
andresecarrasco.blogspot.com
6th
European
Conference
of
GMO-­‐Free
Regions
Brussels
and
Ghent
16-­‐17
September,
2010
ARGE
Gentechnik-­‐frei,
Soy?
–GMO-­‐free
&
Sustainable
Vienna,
April
28
2011

22. Two types of experiments were designed :
1. Embryos were cultured saline solution
(BM) with 1/5000 dilution of GBH (430 uM)
2. 200 to 300 pg of glyphosate (in 4.6nl) was
injected in one blastomere of the two cell
embryo (final concentration in the injected
cell was 10 uM)
4.6nl
Injection volume represent 1/220 fold the
volume of the injected egg (1ul) Izq Der

23. Both GBM and glyphosate
injection alter midline formation
by down regulation of Shh and
Pax6
In tadpoles down regulation of
Otx2 and Sox9 produces:
1. Microcephaly
2. Microphthalmia
3. Craniofacial disruption
(branchial arches)
.
5. Shortening of anterior posterior
(AP) axis.
Paganelli, et al 2010

24. In larvae stage both GBH and
glyphosate produce disruption
of development craniofacial due
to inhibition of regulation of
genes expressed in the
embryonic midline (Shh, Otx2)
1. Microcephaly
2. Microftalmia
3. Craniofacial cartilages
4. Shortening of Anterior posterior
axis.
5. Ciclopia and Holoprosencephaly
Paganelli,
et
al
2010

25. Retinoic acid (a derivate of vitamin
A) plays an important role in the
regulation of expression of genes
controlling early embryogenesis in
vertebrates.
Retinoic acid is an important
morphogen for the pattern
formation of three embryonic axis
• BH increase retinoic acid
G
activity.
• A antagonist (Ro) rescue the
R
GBH induced phenotype.
Paganelli,
et
al,
2010

26. In
conclusion
Synthesis
or
degradaNon
Glyphosate
(RALDH
or
cytochrome
P450
CYP
26
A,
B
y
C)
FGF
8
Anencephaly
Retinoic
acid
HOX
anterior-­‐posterior
axis
(HNF3B)
Cranial
Otx2
Shh
neural
Otocephaly
crest
HPE
and
cyclopia

28. Retinoic acid

29. Retinoic Acid
 RA is a diffusible morphogen present in vertebrates that biding
a family of nuclear receptors (RAR), regulating early genes during
gastrulation (3 week of pregnancy) and late morphogenesis.
 RA regulates formation of AP embryonic axis
 In adults play role in fertility, vision, prevents neoplastic growth,
and neurodegenerative disease.
RA excess cause 20 % of the abortions and 35 % of
the malformations during embryonic pattern
formation

30. RA is ligand for a nuclear receptor and gene regulator

31. Retinoic Acid gradients in the embryonic A/P axis
Pera et al 2009

32. Human syndromes caused by
excess of RA
(excess of vitamin A)

33. CRS: Caudal Regression Syndrome in mice.
(excess of RA) (Padmanabhan R., Retinoic Acid-induced caudal regression syndrome in the mouse
fetus, Reproductive Toxicology, 1998)
Exencephaly. Microcephaly, Microphthalmia. Exophthalmia.
Holoprosencephaly,(HPE) Spina bífida y mielomenigocele
Agnatia or micrognatia, aglosia, palate agenesis, cleft palate craniofacial
abnormalities. Astomia (mouth agenesis) and Anotia (auricular agenesis),
Imperforate anus, Rectouretral fistula , Kidney malformations or kidney
agenesis, Criptoquidia. Gastroschisis,
Reduction ribs number
No tail and caudal vertebral defects,
Limb malformations (meromelia, sirenomelia* (see picture) and
syndactily)
Pathological morphogenesis of heart, lung, kidney, pancreas)

34. Caudal Regression Syndrome could be explained as:
 A disorder of embryonic midline formation
results in the inhibition of several regulatory
pathways (Hox complex, A/P axis, Otx2, Shh, FGF8,
etc.).
 Functional disruption of developmental
mechanisms: proliferation, apoptosis and cell
migration during early vertebrate morphogenesis.

35. Retinoic acid syndrome
Inhibition of Otx2 (forebrain and midbrain)
HPE (ciclopia and prosbosis) + agnatia and NEURAL CREST
sinotia
Inhibition of Shh (embryonic midline) HPE,
NEURAL CREST situs inversus, limbs (amelia, sirenomelia,
sindactily, etc.)
Missregulation of Hox genes (affects A/P axis,
vertebral identity and hindbrain)
Malformations: lung, diafragm, eyes , hearth, digestive tract,
kidney, gonades, etc.

36. Scientific reports associate chemicals and abnormal
morphogenesis for retinoic acid pathway disruption.
Atrazine Lenkowski J, et al. Env Health Persp (2008, 2010)
Tridimefon, Papis E, et al. Gene expression patterns
(2007)
Arsenic, Davey C, et al. Env Health Persp (2008)
Alcohol, Kot-Leibovich H, et al. Dis Model Mech (2009)
Azoles, Giavini E. and Menegola E., Tox. Letters (2010)

37. Glyphosate in mammalian embryos

38. Toxicokinetics of glyphosate
Doses:
IV 100 mg/Kg
Oral 400 mg/kg
Slowly disappearance from blood
Intravenous half life: 10 hr.
Oral half life: 14 hr (degradation to AMPA 6%)
Distribution (two compartment model) both
administrations indicates that glyphosate penetrate easily
all tissues (according with Brewster et al 1991)
Anadon, et al, Toxicology letter (2009)

39. Implantation in humans
7 days
10 days
Human Embryo 14 days
14 days

40. “These experimental conditions do not reflect human exposure, kinetics
(absorption, distribution, excretion and their time course) in the mammalian
organism or factors such as the placenta barrier in the pregnant female. In no
case, exposure of the developing human fetus in utero would be similar to that
one of the frog or chicken embryos in the study by Paganelli et al.” (page 3-4)
Bundesant fur
Verbraucherschutz und
Lebenmittelsicherheit
19 October 2010
Wrong!!!!!!
Placenta is permeable to glyphosate.
15 a 20 % of glyphosate in blood, cross the
placenta barrier.

41. In rats: In mammalians (rats).
Teratogenesis at 3.5 g/kg
(acute oral LD50: 5.6 g/kg)
They used:
1 g/kg
075 g/kg
0.5 g/kg
Administration 9 days/pc
Gastrulation 7 days/pc
Dallegrave et al, 2003
LD50 5.6 g/kg glyphosate Roundup (Brasil)
38% glyphosate More information see:
LD50 1g/kg POEA 18% POEA
“Reply to the letter to the Editor Regarding our article”
Chem. Res. in Toxicology, 24, 2011, 610-615

42. Comments
treatment was done after late gastrulation when the
primitive streak and the axis are already setup.
craniofacial malformations
malformation in the skeleton (vertebrae, ribs, limbs, femur,
etc)
All of these malformations are consistent with the
increase of RA during development.
(see also “Reply to the letter to the Editor Regarding our article” Chem. Res. in Toxicology,
24, 2011, 610-615)

43. Persistency of glyphosate in the soil

44. New evidences of Roundup (glyphosate formulation) impact
on the periphyton community and the water quality of
freshwater ecosystems. Marıa S. Vera, Leonardo Lagomarsino,
Matıas Sylvester, Gonzalo L. Perez. Patricia Rodrıguez, Herna´n
Mugni, Rodrigo Sinistro, Marcela Ferraro, Carlos Bonetto, Horacio
Zagarese, Haydee Pizarro
Ecotoxicology (2010) 19:710–721
“Unexpectedly, we detected that a single application of glyphosate in
2005 shifted the mesocosms from a ‘‘clear’’ to a ‘‘turbid’’ state which
remained until the next year. As was discussed above, the glyphosate
may be adsorbed to sediments and a slow later desorption might
produce a long turn effect suppressing growth of the most sensitive
groups and favoring the abilities to compete of the more resistant
algae. This trend in long term effect was suggested by Holtby and
Baillie (1989) who reported an enhancement of periphytic production
as a response to increased levels of phosphorus produced by a unique
application of Roundup done 1 year before their experiment, carried
out in natural streams”

45. GENOTOXICITY

46. GENOTOXICITY (Comet assay )
Type A (normal)
Type C
Type E
DNA Fragmentation
Type B Type D Type F

47. Genotoxicity (comet assay) (Rio Cuarto, CORDOBA)
Mañas y Aiassa 2010

48. PARAGUAY

49. PARAGUAY
Benitez Leite 2010