Documente Academic
Documente Profesional
Documente Cultură
Asesor
Ral Gerardo Can Suarez, Docente Facultad de Ciencias Econmicas
Dedico esta obra a mi hijo, Erik Martin Colmenares Velasco, a mi prometida Vivian Katerine
Velasco Aldana y a mis padres, quienes han inspirado mi investigacin y dedicacin constante a la
sublevacin del consumo desproporcionado y mundano del mundo actual, al cuidado de la
naturaleza sobre los intereses individuales, al profundo amor por mi nacin y todo lo que significa
tan majestuoso significado, en ndole insuperable, dedico esta obra a aquellos que prefieren no
guardar silencio para no ser sentenciados como inocentes o en su defecto, muertos como borregos.
Agradezco a mi ncleo familiar, a mis maestros y docentes, por ser quienes han
dispuesto de su sacrificio profesional y humano, para hacer de m un hombre recto y
verdadero, adems de mostrar el camino del aprendizaje constante, la
autodeterminacin y la ansias de romper los esquemas, con el fin de generar un criterio
propio como evidencia de nuestro objetivo terrenal, ser humano.
TABLA DE CONTENIDO
TITULO ...................................................................................................................................... 7
DEFINICION DEL PROBLEMA............................................................................................. 7
JUSTIFICACION ...................................................................................................................... 8
OBJETIVO GENERAL ............................................................................................................ 9
OBJETIVOS ESPECFICOS .................................................................................................. 9
HIPOTESIS ............................................................................................................................. 10
METODOLOGIA ................................................................................................................... 10
CAPITULO 1: LA RESPONSABILIDAD SOCIAL EMPRESARIAL COMO PUNTO
DE PARTIDA .......................................................................................................................... 12
PRINCIPIOS DE LA RESPONSABILIDAD SOCIAL ........................................... 12
La Ciudadana Corporativa ............................................................................................................... 14
STAKEHOLDERS ..................................................................................................... 20
La teora de los stakeholders ........................................................................................................... 21
Los intereses de los stakeholders .................................................................................................. 23
Los intereses de tipo material ........................................................................................... 24
Los intereses polticos ........................................................................................................ 24
Los intereses de afiliacin .................................................................................................. 24
Los intereses referentes a la informacin ...................................................................... 25
Los intereses de lineamiento simblico ......................................................................... 25
Los intereses de tipo metafsico o espiritual................................................................. 25
TITULO
En 1901 en San Lus, Misuri, Estados unidos; John Francis Quenny, un qumico de
la industria farmacutica, fund la empresa Monsanto, nombre que recibi del
apellido de soltera de su esposa, Olga Mndez Monsanto.
Comenz su funcionamiento con capital propio y con distribucin de sacarina; a
travs de los aos y gracias a su crecimiento econmico se convirti en la
transnacional de insumos agropecuarios ms grande del mundo; especializada en la
creacin, produccin y distribucin de semillas modificadas genticamente;
actualmente, Monsanto tiene distribucin en el mercado mundial de sus productos y
como es de esperarse, tambin se encuentra en el mercado nacional colombiano, la
adquisicin de sus productos por parte de los agricultores colombianos, sin un
estudio riguroso de los insumos, frente al consumo de lo que se considera orgnico,
convirtindose en una amenaza directa para las semillas tradicionales, los intereses
de la poblacin rural, el consumo de la sociedad civil, las tradiciones alimentarias y la
salud.
Para tal fin se considera que un anlisis de la corporacin Monsanto es de gran
importancia para los consumos alimentarios de la nacin, tomando como referencia
su ascenso, sus productos y la responsabilidad social empresarial, que hace de esta
empresa una de las compaas ms crecientes del mundo con dudosa reputacin
internacional.
Por qu Monsanto es una empresa tan cuestionada internacionalmente?
JUSTIFICACION
OBJETIVO GENERAL
Generar
argumentos
adicionales
al
planteamiento
actual,
referente
la
OBJETIVOS ESPECFICOS
HIPOTESIS
La transnacional Monsanto maneja ms del 60% del mercado mundial de
productos genticamente alterados; un ejemplo de ello es la soya, producto que
en estados unidos equivale al 90% de su produccin total como nico del
desarrollo transgnico y que es encontrado en la mayora de alimentos vendidos
para el consumo de la poblacin civil en general. Los organismos genticamente
alterados estn en su apogeo, pese a que son ms duraderos y resistentes en la
siembra y cosecha que las semillas orgnicas; los productos transgnicos han
desencadenado fuertes contradicciones y apoyo en los hbitos de consumo
mundial; por lo que es necesario dar un vistazo ms profundo a Monsanto, ya
que de ser la alimentacin transgnica el futuro alimentario mundial, no se puede
reducir el grado de importancia para su aprobacin en general, teniendo en
cuenta que Monsanto ha efectuado millonarias inversiones a los OMG y que
adems, muestra fuertes nexos con los entes de regulacin y los gobiernos en
los pases destino de sus productos, por lo que amenaza directamente los
intereses generales de la ciudadana y que suspicazmente alinea poltica y
econmicamente su entrada a los pases anfitriones de sus alimentos
favorablemente resistentes, aunque, la historia de esta gran organizacin mundial
oculta un rostro que ha salido en ocasiones a contradecir con gran complicidad
de los aliados de la compaa, que Monsanto guarda ms de un pernicioso
secreto .
METODOLOGIA
Descriptiva: se partir de los conceptos de la responsabilidad social empresarial,
escenarios estratgicos, stakeholders, con el fin de identificar la RSE en
Colombia; adems, generar contenido con la recopilacin de informacin
concerniente a la empresa Monsanto a nivel mundial a travs de:
1. Rastreo de informacin de la empresa a nivel mundial, su modo de
operacin para entrar a mercados extranjeros, los productos ms
10
construccin
de
un
documento
que
permita
identificar
las
11
12
contemplar los aspectos de la estrategia. Por otra parte Wood (1991) define: la
disposicin de una organizacin empresarial con principios de responsabilidad social
est compuesta por, procesos de respuesta, las polticas, los programas
contemplados, y los resultados observables de las relaciones sociales que pretende
(p.693), Wood expone que los principios de la responsabilidad empresarial, los
procesos, programas y anlisis son aquellos que conforman la RSE. Es decir, la
idea de la responsabilidad social empresarial ha sido ampliada progresivamente, en
donde tambin se han acuado expresiones para dar cabida a nuevos conceptos.
Waddock (2004) acua el concepto de Ciudadana Corporativa, en donde permite
incorporar a la responsabilidad social empresarial con sus principios fundamentales,
en donde se consolida la necesidad de incorporar estrategias y polticas dentro del
proceso general de gestin en las organizaciones, permitiendo la idea de relacionar
los diferentes pblicos de la organizacin como parte de desarrollo del proceso de la
ciudadana corporativa. La responsabilidad social empresarial ha tenido un cambio
ajustado a los intereses del bien comn, es decir, el productor dispone de recursos y
equipos para realizar un producto, adems de su inversin, pero el si el mismo no
contempla que se est usando un recurso escaso de la naturaleza o en su defecto
daa el ecosistema para la realizacin de un producto, el consumidor reaccionara
negativamente a los estmulos de comercializacin, por lo que los hbitos de
consumo se han favorecido de la responsabilidad de cada poca y de los mbitos
que lo rodean, como ejemplo claro de este paradigma esta la finalizacin de la
segunda guerra mundial, pese a que el auge industrial de la poca incremento
13
La Ciudadana Corporativa
ESCENARIOS ESTRATGICOS
Los escenarios de las empresas, son parte del anlisis continuo de la llamada
frontera entre la empresa y el entorno; mencionada frontera es la divisin terica de
lo que significa responsabilidad para el empresario, segn la tendencia que se
14
la eleccin de
Cuando
se
contempla
un
escenario
estratgico,
se
est
reuniendo
15
Con el fin de identificar una comprensin contextual de los escenarios sobre los
cuales despus se actuara, es de gran importancia antes, considerar los
lineamientos de anlisis. Garrido (2008) expone; los limites, establecen las fronteras
en torno al eje del cual se llevar el anlisis, lo que permite situar a los actores que
interferirn en el proceso y su alcance; el escenario y contexto, se identifican las
interrelaciones del anlisis, con el fin de aislar en detalle, la relacin
tensin/atraccin sobre la que se lleva el escenario; actuacin, identificar los actores
reales del proceso, en donde est la visin del estratega y la visin de los clientes,
determinando una realidad conjunta de los actores que intervienen en el proceso; la
interaccin, la identificacin y descripcin detallada del perfil de los actores, orientar
respecto a la significancia o insignificancia que los actores tengan para la
consecucin de la estrategia; la adaptacin, a partir de los indicadores obtenidos
desde los anlisis del optimo en los escenarios estratgicos en estado futuro, se
deben generar las formas de adaptacin de procesos y operaciones que aseguren la
rentabilidad y el retorno de la inversin.
16
Responsabilidad econmica
Responsabilidad legal
18
Responsabilidad filantrpica
Responsabilidad medioambiental
19
STAKEHOLDERS
20
la teora de los stakeholders constituye una feliz alusin que puede cambiar la
percepcin, hacia a una situacin en que las empresas y el propio sistema capitalista
contribuyan a configurar una realidad ms justa y beneficiosa para los interesados
de la compaa; quizs para los directamente implicados de la actividad econmica
(inversores y directivos), manifiestan una antropologa ms simplista, en donde el
valor real est en apropiarse del valor econmico y no de crearlo, se sobrepone la
importancia de la competicin, la escases de la materia prima, el juego de suma cero
y el hecho de que el ganador es quien se queda con todo.
Siendo as, no hay tiempo para la tica. Sin embargo, no tiene porqu ser as, no
se trata de redefinir los conceptos de stakeholders ante el capitalismo, si no, de
hacer de este un concepto til realmente.
La controversia en la teora de los stakeholders se encuentra en los aspectos
diferentes en la teora: el descriptivo, el instrumental y el normativo. El aspecto
21
que ejerce
la
compaa, adems de
afectado directa e
22
24
cosas tales como, los valores que promulga la empresa, y si se alinean con los
intereses de la comunidad local en donde se desarrolla la actividad econmica.
25
conlleva
competitividad,
sostenimiento
desarrollo
de
la
26
27
28
Un pas de
29
CAPITULO 2: MONSANTO
QUIN ES MONSANTO?
30
esperarse. Robin (2010) menciona que algunos cientficos alertaron al pblico sobre
los riesgos para la salud, excluyendo as la aprobacin de sus colegas para el
consumo de los alimentos transgnicos; indicados cientficos enfrentados a la
desaprobacin y a represaras en sus lugares de trabajo, abrieron las puertas al
debate nacional sobre las irregularidades del funcionamiento de Monsanto en un
pas en donde la poblacin desacreditaba sus productos; los cientficos demostraron
que el herbicida Roundup es sumamente toxico, aunque se vendiera como
biodegradable; Monsanto fue condenada por publicidad engaosa y fraude, para el
2009 Monsanto pago una multa de 15000 euros y se le condeno a dejar de utilizar
dicha publicidad del producto. A partir de all surgieron preocupaciones sobre el
riesgo de contaminacin de los cultivos transgnicos a plantaciones orgnicas; la
desaparicin de las pequeas granjas,
Prdida de la autosuficiencia alimentaria y la destruccin de procesos tradicional
de produccin de alimentos. Con tal contexto, el concepto de la soberana
alimentaria de los campesinos franceses, fue el concepto que avivo los movimientos
que desechaban los productos transgnicos de Francia, Alemania, Espaa, Austria.
arrojados al canal de la ciudad por parte de la compaa, fue la principal causa por lo
que los habitantes de Anniston se vieron directamente afectados, para cuando fue
expuesto al pblico el desastre, tuvieron que desalojar sus casas y abandonar estos
sitios pese a la alta toxicidad del qumico registrado en el ambiente, lo cual hacia
inhabitable las zonas en donde se hizo el estudio toxicolgico, para los habitantes
fue muy tarde en la mayora de los casos, pese a que la mayora ya haban tenido
una exposicin directa al qumico, por lo que se vieron afectados con cncer,
hepatitis, diabetes, malformaciones congnitas y por ltimo la muerte (Robin, 2010).
El ecologista ken Cook expone durante su investigacin del PCB, producto
empleado por Monsanto, que estos estaban al tanto de las implicaciones negativas
al estar en contacto con el qumico; en 1937 se exponen pblicamente los efectos al
PCB (cncer, efectos txicos en los rganos, erupciones cutneas, hepatitis); en
1961, dos obreros expuestos a la ruptura de un tubo dentro de la empresa, tubo que
contena el qumico, comenzaron a
Roundup
36
Es decir, en Estados Unidos, los ministros no dan la talla ante los intereses de las
multinacionales; la FDA, es la agencia encargada de controlar los productos para el
consumo de los estadounidenses, el 29 de mayo de 1992 con la resolucin de los
alimentos transgnicos se tom la decisin de no crear una categora especial para
los alimentos genticamente alterados, al contrario, se les clasifica en la misma
actividad de la cruza de especies tradicional. James Maryansky (coordinador
biotecnolgico de la FDA), en ese entonces diriga el departamento de biotecnologa;
Maryansky (2010) dice en la entrevista que hace Marie-Monique Robin en su
documental bsicamente el gobierno, tomo la decisin de no crear leyes nuevas,
pensbamos que las leyes existentes podan aplicarse a las nuevas tecnologas. Es
donde la casa blanca escribi un informe a la FDA, con el fin de que los
transgnicos, no tuvieran un trato diferente a los productos que eran cultivados
tradicionalmente. Robin (2008) afirma que esta decisin no se tom con
fundamentos cientficos, sino como una decisin poltica que congregaba a muchos
alimentos en la decisin, no solo a las plantas, sino a todo producto manipulado con
biotecnologa para el consumo humano, lo que la FDA llamo el principio de
equivalencia
sustancial;
El
principio
de
equivalencia
sustancial
expresa
deliberadamente que tanto los alimentos orgnicos como los transgnicos deben ser
tratados de igual manera, sin distincin alguna de los que son cosechados
orgnicamente con aquellos que son alterados genticamente, la FDA muestra a los
productos transgnicos como iguales con los de uso tradicional. El principio de
equivalencia sustancial, fue la justificacin para que las empresas como Monsanto,
sacaran al mercado rpidamente sus productos sin controles estrictos para el
consumo, esto efectuado con la mayor injerencia gubernamental.
Samuel Epstein, es el dirigente por la coalicin de la prevencin del cncer; en
1990, el profesor Epstein pblico un artculo en la revista MilkyWeek, despus de
recibir una informacin de una fuente annima, informacin que contena, datos
confidenciales de la FDA, acerca de los productos producidos por Monsanto, publico
una primicia basada en estos documentos e informacin recaudada por el profesor
minuciosamente, esta caja que recibi contena los estudios veterinarios que
Monsanto habra enviado a la FDA, desde haca 6 aos; los documentos fueron
37
38
El agente naranja
39
momento James Maryanski supo que la EMS era provocada por tal OMG y no
reporto a los entes gubernamentales encargados.
En el ao 1998 se lleg a conocer otro caso que tena como autor a la
multinacional Monsanto, el cientfico Arpad Pusztai, un estudioso independiente, que
pese a sus investigaciones perdi su empleo, como se ha mencionado en anteriores
casos; despus de demostrar sus hallazgos ante el instituto Rowett de Aberdeen,
Escocia; en su investigacin el Dr. Pusztai, muestra que las papas o patatas
transgnicas, tenan efectos colaterales; para el perodo, Gran Bretaa se
encontraba entusiasmada con la entrada de los OMG al continente, Pusztai y sus
colaboradores demuestran que los OMG, provocaban una proliferacin de clulas en
el estmago de las ratas con el cual llevaron su experimento, lo cual no es bueno;
este hecho genera tumores cancergenos causados por productos qumicos, el
sistema inmunolgico de los roedores se alteraba y el organismo de las ratas trataba
a las patatas transgnicas, con las que fueron alimentadas, como cuerpos extraos y
no como parte de la dieta, al ser consumida. Se demostr con ello que el causante
directo de tales malformaciones cancergenas eraprovocado por la alimentacin a
base de OMG. Al ser expuesto el estudio del Dr. Pusztai, este fue despedido junto a
su equipo de trabajo, por parte de los directivos del instituto Rowett. En una de sus
afirmaciones, Pusztai menciona que los OMG son organismos que no tiene un
control en las generaciones futuras, adems que as como ocurri con las ratas,
estas
mencionadas
malformaciones
cancergenas
son
heredadas
durante
generaciones, siendo cada vez ms agresivo. Pusztai (2008) sera muy injusto, muy
injusto, usar a nuestros conciudadanos como cobayas para los organismos
genticamente modificados Los cientficos y colaboradores que trabajaron con
Pusztai fueron despedidos y desacreditados.
42
Monsanto, al ser una Transnacional, con el poder que manifiesta en los crculos
econmicos estadounidenses, tienen leyes que protegen su actividad; la cuestin de
las patentes, es uno de los factores ms irrelevantes e incoherentes del buen
administrar que expresa Monsanto, Monsanto reclama las patentes de sus productos
como derechos de propiedad intelectual, con lo cual pretende proteger sus
inversiones; en estado unidos todo granjero que use OMG de Monsanto, est
obligado a firmar un documento de empleo de la biotecnologa, el cual compromete
al granjero a respetar la patente que la compaa ejerce sobre el gen manipulado.
Esta ley de patentes prohbe guardar granos para sembrarlos luego; Cmo controla
Monsanto que no se cultiven sus semillas?, Monsanto expone que la empresa se
har cargo de llegar a un acuerdo en los casos en donde los cultivos que puedan ser
expuestos a los OMG de la empresa, con aquellos de los cultivos tradicionales, este
acuerdo por parte de la empresa beneficiara los intereses del granjero anfitrin y de
la compaa. Una idea que parece idnea, pero que en mltiples ocasiones es
desastrosa para los agricultores; aquellos sembradores que son demandados por la
compaa al no respetar sus patentes de semillas, son llevados a procesos legales y
a numerosas quiebras en muchos de los casos. Un ejemplo especifico es una de la
familias que aun siembra soya orgnica del 90% de la soya transgnica que es
comercializada en estados unidos, la familia Rush estuvo expuesta a las fauces de la
multinacional.
El caso de Troy Rush un agricultor de indiana, comienza en 1999; un detective
privado contratado por Monsanto llega a las 19:00 a la granja Rush y les expone a
sus propietarios que est en bsqueda de sospechosos por haber guardado las
semillas de la compaa, los Rush mencionan al investigador que sus semillas han
sido adquiridas por medio de un distribuidor que no tiene nada que ver con
Monsanto, los herbicidas que adquieren en la granja tambin se hace por medio del
mismo proveedor. Dos semanas despus reciben una notificacin de la demanda
por las patentes, en donde Monsanto expone que los Rush han usado sus semillas,
43
MONSANTO EN EL MUNDO
viento, manipulacin humana de la planta; es por ello que se expone, prohibir la miel
que ha sido encontrada con polen de plantas transgnicas, con la finalidad de evitar
el consumo de transgnicos por parte de los pases que reglamentan su prohibicin
justificada.
En India, en febrero del 2010, el gobierno enfrento la oposicin masiva, entre
agricultores, ciudadanos y cientficos, preocupados por los impactos negativos de la
berenjena Bt de Mahyco-Monsato sobre los medios de la vida y la salud humana, las
autoridades ejercieron una moratoria para este OMG, en donde la soberana
alimentaria ganaba adeptos del pas asitico; para agosto del 2011, la Autoridad
Nacional de Biodiversidad, presento una demanda contra Mahyco-Monsanto y a sus
colaboradores por biopiratera. La empresa uso seis variedades de berenjena nativa
en el desarrollo de una versin transgnica del cultivo.
45
En Mali,
los
cultivos
transgnicos
son
constantemente
mencionados
en
46
MONSANTO EN COLOMBIA
47
identificar la comida orgnica de los OMG, ley que en otros pases, obliga a
discriminar los alimentos transgnicos de los puramente orgnicos.
Es decir, el
plazo.
El ICA como forma de proteccin al consumo de los productos autctonos y
ancestrales, prohbe la siembra de transgnicos en resguardos a una distancia no
menor a los 300 metros, sin contemplar que diversos estudios de la reproduccin
vegetal demuestran que la contaminacin gentica del maz transgnico corre por
medio del viento con rea no contemplada en metros si no a kilmetros de distancia,
adems de ello el intercambio de productos entre indgenas y la sociedad civil, en
donde la produccin a nivel campesina de los transgnicos no ha sido prohibida y
fcilmente se puede contaminar la semilla no alterada. En el pas se habla muy poco
acerca de las leyes que protegeran la contaminacin de los productos de consumo
para las poblaciones ms vulnerables. Frente a esta problemtica en el 2008 se
interpuso una demanda contra el ICA, alegando all todas las decisiones frente al
uso de organismos genticamente alterados, debe ser consultado con el pblico en
donde este principio nombrado en el protocolo de Cartagena an se encuentra en
proceso. Una de las situaciones ms delicadas en el actual curso de la entrada de
productos transgnicos al pas, fue la falta de informacin por parte de los
organismos gubernamentales que quienes sin una consulta popular han aprobado el
consumo de productos sin control riguroso de las consecuencias que pueden
generar.
Monsanto as como otras empresas congregadas a la produccin de productos
transgnicos, sostienen que estos cultivos son la solucin al hambre, al
almacenamiento de carbono y a efectos de los cambios climticos, a pesar de todos
los anlisis han fracaso una y otra vez (Acua, 2005). Estos anlisis muestran que
los
productos
transgnicos
son
iguales
en
rendimiento
los
productos
48
como
alimento
entonces
puede
ser
beneficioso?
Cuando
las
49
50
51
52
53
54
Monsanto puede ser la industria del milenio, generando respeto, dignidad para el
consumo, empleo digno, solidaridad, contribucin al bien comn, corresponsabilidad,
confianza, tica empresarial, transparencia, honestidad, legalidad, desarrollo social;
la responsabilidad social aplicada a Monsanto sera un gran ejemplo de compaa
realmente responsable con el mundo, actualmente para aquellos conocedores, lo
nico seguro de la compaa, es que, algo oculta que no puede revelar pese a la
tenacidad de sus repercusiones.
Monsanto supone que el terreno en el que sus cultivos crecen, tendrn una
productividad limitada, por lo que los agricultores tendrn que migrar sus cultivos del
punto de origen, esto pese a que el glifosato altera los nutrientes de la tierra virgen
de OMG, es decir, habra que esperar a que el glifosato se diluya totalmente de la
tierra para tratar de recuperar la tierra. Pese a que las malas hierbas bloquean el sol
directamente sobre la tierra, estas malas hierbas se desintegran y aportan nutrientes
importantes para el cultivo de nuevos cultivos orgnicos; al aplicar glifosato en un
cultivo, este elimina esas malas hierbas que bloquean el sol de manera ms rpida,
exponiendo a la tierra al calor directo del sol, secando la tierra y dejndola estril a
largo plazo, sin la posibilidad de recuperar esos nutrientes al estar contaminados de
glifosato durante generaciones de cultivos venideros, secando la tierra de nutrientes
y del agua subterrnea que tambin lentamente es envenenada por las filtraciones
de partculas de glifosato, dejando a largo plazo una parcela de tierra sin alguna
probabilidad de siembra para aprovechamiento en el futuro de la misma. Monsanto
es consciente de estos efectos y no ha revelado esta informacin; esto pese a que
es prohibido realizar estudios de sus productos para revelarlos a la luz pblica; sin
mencionar los casos medioambientales de gran envergadura en los que han estado
involucrados sus productos, como Vietnam, Anniston, India, Francia, Argentina,
E.E.U.U., UE, frica, Suiza, Mali, etc.
55
CONCLUSIONES
57
REFERENCIAS
59
Zacune, Joseph. & recopilacin activistas de todo el mundo (2012). Lucha contra
Monsanto: Resistencia de los movimientos de base al poder empresarial del
agronegocio en la era de la economa verde y un clima cambiante. La va
campesina, amigos de la tierra internacional, Combat Monsanto 12(1), 5-20.
Zadek, S. (2005). El camino hacia la responsabilidad corporativa. Harvard Business
Review Amrica Latina.
ILUSTRACIONES
ILUSTRACION 1: aspectos de la ciudadana corporativa
Accin econmica
CORPORATIVO
Accin social
Accin medioambiental
Gobierno corporativo
TICA CORPORATIVA
tica de negocia
RELACIONES CORPORATIVAS
60
ANEXOS
61
las entreguemos a las generaciones futuras. Para las comunidades campesinas, indgenas y afro
colombianas las semillas han sido el fundamento de su cultura, de sus sistemas productivos. Es
por ello que los agricultores tienen el derecho al libre acceso, a la produccin, a guardar,
intercambiar y vender las semillas. Las semillas nativas y criollas, se constituyen en el seguro que
tiene la humanidad para enfrentar la profunda crisis de la agricultura frente al cambio climtico.
Las semillas son la base de la libertad de los alimentos, ya que son el primer eslabn en la
cadena alimentaria. Sin semillas libres de propiedad intelectual y sin el control local de sus
territorios, no es viable la soberana y autonoma alimentaria de toda la poblacin y tampoco es
posible que las comunidades vivan dignamente en el campo y en paz. La libertad de las semillas
se ve amenazada por la biopiratera y las patentes que crean monopolios y vuelve ilegal que los
agricultores guarden e intercambien sus semillas; y tambin por las semillas transgnicas que
contaminan nuestros cultivos, cerrando as la opcin de alimentos libres de transgnicos.
En Colombia las leyes que vulneran la
Libertad de las Semillas son:
- El artculo 4 de la Ley 1032 de 2006 que criminaliza a los agricultores por sembrar semillas de
las empresas sin autorizacin y tambin las semillas similarmente confundibles a una semilla
protegida legalmente.
- La Resolucin 970 de 2010 del ICA, es un instrumento para perseguir, volver ilegal y
criminalizar el uso de semillas nativas y criollas por los agricultores y exige que solo se pueden
utilizar semillas certificadas y registradas.
- Ley 1518 de 2012 que aprueba el Convenio para la Proteccin de las Obtenciones Vegetales
UPOV 91, le permite a las empresas apropiarse de las semillas, puesto que define como
obtentor a La persona que haya creado o descubierto y puesto a punto una variedad;
permite una proteccin similar a una patente y desconoce los derechos de los agricultores al
impedirles la resiembra, uso, intercambio y comercializacin.
-El Decreto 4525 de 2005 de bioseguridad, permite sin los debidos controles, la liberacin
comercial de cultivos y alimentos transgnicos y la contaminacin de las semillas nativas y
criollas.
Las organizaciones locales y sociales de Colombia consideramos que estas leyes y normas de
semillas son ilegtimas e inconstitucionales, por que violan los derechos humanos fundamentales
de las comunidades rurales, los convenios internacionales suscritos por Colombia, como el
convenio 169 de la OIT y las leyes que protegen los derechos delas comunidades tnicas y la
soberana alimentaria nacional. Igualmente violan los derechos que tenemos los colombianos
sobre el patrimonio gentico de la nacin y los consumidores urbanos perdemos los derechos a
una alimentacin sana y biodiversa que nos proporcionan las semillas que estn en manos de los
agricultores.
Por todos los argumentos expuestos, las organizaciones locales y sociales de
Colombia, manifestamos:
Nuestro Rechazo al control corporativo sobre la vida, a todas las formas de propiedad intelectual
sobre las semillas, a los organismos vivos y su conocimiento asociado, la biopiratera, las semillas
62
transgnicas, las leyes ilegitimas que criminalizan el libre flujo y multiplicacin de las semillas.
Estas normas violan los derechos sobre el patrimonio gentico y cultural de los colombianos y
especialmente los derechos colectivos las comunidades indgenas y campesinas sobre su
biodiversidad y tambin el derecho de los ciudadanos a una alimentacin libre y soberana.
Consideramos ilegales todas las normas que operan en Colombia que pretenden regular el uso
de semillas, en el marco de la UPOV 91, la decisin 345 de la CAN, el artculo 4 de la ley 1032 de
2006, y la resolucin 970 de2010. De otra parte, todas estas normas han sido proferidas sin
realizar la consulta previa libre e informada con poblaciones tnicas. No vamos a reconocer
ninguna ley que otorga de manera ilegtima la propiedad privada de las semillas y el control
monoplico a las empresas de las semillas, puesto que estas son patrimonios colectivos de los
pueblos.
Rechazamos los decomisos de semillas y judicializaciones a agricultores que el Instituto
Colombiano Agropecuario ICA est realizando en diferentes regiones del pas. Entre 2010 y
2011 se decomis 1.167.225 kilogramos de semillas. Les notificamos que por cada semilla que
nos decomisen, haremos que estas germinen y florezcan de nuevo, se multipliquen, se
esparzan y caminen libremente con los agricultores por los campos de Colombia.
Saludamos el proceso de negociacin que actualmente adelanta el gobierno colombiano con las
Farc y el Eln y esperamos que finalmente se pueda acabar con esta cruel guerra que tanto dao
le ha costado al pas. Creemos que la paz solo se lograr si existe la voluntad poltica de todas
las partes para realizar cambios que resuelvan problemas estructurales, en donde se involucren
los acuerdos de paz especialmente a la poblacin afectada por la guerra.
No es fortuito que uno de los temas inciales y centrales en la agenda de negociacin actual, sea
el tema agrario y de tierras. Consideramos que entre los temas centrales en la negociacin se
debera incluir una verdadera reforma agraria, que permita a las poblaciones campesinas,
indgenas y afro vivan dignamente y en paz en sus territorios; y una poltica de fortalecimiento
de la soberana y autonoma alimentaria para el campo y la ciudad. En este contexto, se deben
reconocer los derechos colectivos y el control local de los territorios, que incluya los recursos
naturales all existentes, la tierra, el agua, los bosque y las semillas como fundamento de los
sistemas productivos de la agricultura familiar y comunitaria. Es por ello que consideramos que
el tema de las semillas debera ser incluido en el actual procesode negociacin de paz.
Exigimos polticas pblicas a favor de sistemas vivos de semillas campesinas, que estn en
nuestras comunidades y bajo nuestro control, que promuevan las semillas reproducibles y
locales, que favorezcan la agro ecologa, el acceso a la tierra y el cuidado de los suelos, y
tambin deben promover la investigacin participativa controlada por los campesinos. Estas
polticas deben prohibir la privatizacin y acabar con los monopolios de las semillas por la
industria y no deben promover las semillas no reproducibles como los hbridos y tampoco los
transgnicos.
Hacemos un llamado a las comunidades indgenas, afro y campesinas de Colombia a conservar,
proteger, desarrollar y compartir nuestras semillas; esta es la mejor forma de resistir contra el
despojo y la mejor forma de preservar la biodiversidad. Continuaremos trabajando dentro de
nuestros propios sistemas campesinos de semillas, los cuales han asegurado la biodiversidad y
alimentado a la humanidad y lo continuarn haciendo ante la crisis climtica actual.
63
Igualmente llamamos a la ciudadana en general que habita las ciudades, para que realice
acciones por la defensa del derecho a la alimentacin y reivindique la soberana alimentaria
nacional. Actualmente, las comunidades rurales en Colombia producen ms del 60% de los
alimentos que se consumen en las grandes ciudades; en ese sentido, la privatizacin y
penalizacin del uso de las semillas, pone en riesgo el derecho a la alimentacin de los
colombianos.
Las semillas en manos de los agricultores son un eslabn fundamental para que las poblaciones
rurales y urbanas garanticemos nuestra soberana y autonoma alimentaria. Es por ello que los
agricultores estamos guardando las semillas, no solo para nosotros sino tambin para nuestros
hijos; puesto que las semillas como lo reafirma la Va Campesina son patrimonio de las
comunidades campesinas y afro colombianas y de los pueblos indgenas al servicio de la
humanidad.
abstract
Polychlorinated biphenyls (PCBs) are
synthetic organochlorine chemicals that were
useful industrial products in the past, but
their production was ended because they
persist in both the environment and living
organisms. The PCBs are mixtures of up to
209 different com-ponents (congeners),
depending on the number and position of
chlorines around the biphenyl ring. The
PCBs are fat-soluble substances to which
everyone is exposedthrough ingesting animal
fats, inhalation, or dermal contact. Exposure to PCBs suppresses the immune
system, thereby increasing the risk of
64
_________________________________________
Reprint requests to: D.O. Carpenter, School of Public
Health, Department of Environmental Health Sciences, 5
University Place, A217, RensselaerNY12144, USA
e-mail: Carpent@uamail.albany.edu
BACKGROUND
meta ortho
2'
3'
para
4'
meta ortho 6'
5'
POLYCHLORINATED BIPHENYL
Polychlorinated Biphenyl
Fig. 1: Chemical structure of the PCB molecule. The 10 positions are numbered 2 to 6 on one ring and 2 to 6 on the second ring.
Chlorines can be substituted for hydrogen at 10 possible sites on the biphenyl rings. If no chlorines are present at ortho positions
(2,2,6,6), then the molecule exists primarily in a planar form and has dioxin-like activity. If two or more chlorines are present at
ortho positions, then the molecule assumes a three-dimensional configuration. Theoretically, 209 PCB congeners can be formed,
depending on the number of chlorines and their location.congener. For example, congeners containing few chlorine atoms are in
general more water soluble, more volatile, and more easily metabolized than are those having a high number. Conversely, PCBs
with large numbers of chlorines are resistant to biodegradation and therefore bioaccumulate in the environment. Congeners
containing chlorine only in the meta and para positions tend to assume a planar configuration and show dioxin-like activity,
whereas those having more than one ortho chlorine do not show significant dioxin-like activity.
The toxicity of PCB congeners that exist in a planar configuration occurs through the activation of a
protein called the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factorthat controls the
expression of cytochrome P450 1A (CYP1A) genes. The AhRis a member of the steroid-hormone receptor
family and acts by gene induction, increasing or decreasing the levels of a large number of gene products.
Unlike the steroid-hormone receptors, however, the AhRhasno known physiological ligand.The known AhR
ligands are foreign planar aromatic compounds, such as certain polycyclic aromatic compounds and
halogenated aromatic compounds like the dioxins. Thus, the actions of the coplanar PCBs on the AhRare
identical to those of the dioxins and furans.
Four coplanar PCB congeners having at least four chlorines each, as well as several monoorthocongeners, have significant AhR-activating potential. As all chemicals that activate the AhR have similar
actions but different potencies, a toxic equivalent factor (TEQ) (toxicity of a particular formulation
relativeto2,3,7,8,-tetra-chlorodibenzo-p-dioxin [TCDD]),was developed as a way to deal with mixtures of
compounds that act on a common receptor /12/. Although the coplanar PCBs are not as potent in activating
the AhRas TCDD, the most toxic dioxin congener, in many circumstances the concentration of the PCBs is
sufficiently greater than that of dioxins and furans, such that most AhR receptor activity comes from PCBs
/3/.
No person exposed to PCBs is exposed only to dioxin-like PCB congeners. Because the non-dioxin-like
PCB congeners have a different mecha-nism of action and because exposure to such congeners results in
different diseases, a PCB-exposed person is at risk for all diseases caused by dioxin, as well as for those
caused by non-dioxin-like congeners. Each individual congener has its own profile of actions in biological
systems. Thus, although having the measurements of individual congeners is important, we do not always
have adequate information on the actions of each individual congener /4/.
Various investigators have proposed grouping PCB congeners according to their degree of chlorination,
pattern of enzyme induction, or endocrine-disrupting activity /3, 56/. Different PCB congeners induce
different cytochrome P450 (CYP) isoenzymes, which are responsible for the oxidative metabolism (Phase I,
66
activation) of many drugs, steroids, and carcinogens/78/. Dioxins and coplanar PCBs induce the activity of
cytochromes P450 1A1, 1A2, 2A1, and 1B1 (numbers indicate the family, subfamily, and gene). These actions
result in the proliferation of endoplasmic reticulum in the liver, resulting in an increase in liver size and an
alteration in many, if not all, aspects of liver function, including the perturbation of endocrine function. The
immune system is altered and the size of the thymus gland is reduced. Fitzgerald et al. /9/ demonstrated that
individuals with high exposure to nine mono- or di-ortho-substituted PCB congeners metabolize caffeine
more rapidly than do those with low exposure, a measure of CYP1A2 activity in the liver. These observations
demonstrate that exposure to PCBs alters liver function in ways that can affect the metabolism of other
substances, through the induction of P450s, in this case CYP1A2, which is involved in the metabolic
activation of certain carcinogens.
Other congeners cause the induction of CYP 2B1 and 2B2, a pattern similar to that induced by
phenobarbital. These congeners have carcinogenic, neurotoxic, and endocrine disrupting actions that differ
from those of the dioxin-like congeners. A third major group of congeners have activities at both sites and are
called mixed congenersmono-ortho congeners that activate both CYP1A and CYP1B enzymes, as well
as CYP2B. This group contains nine major congeners and contributes significantly to the total TEQ because
some are present in relatively high concentrations /10/. Some congeners induce CYP3A enzymes, whereas
others do not induce any P450s but can act at other sites /7/.
Congeners containing ortho chlorines, but not having AhR activity, have a different profile of actions and
health effects, many unrelated to any P450 enzyme. Such PCBs alter the cells of the nervous /1113/ and
immune systems /1415/, causing a relatively rapid cell death that results from disruption of the membrane
structure /16/, an effect not seen by coplanar PCBs at comparable concentrations. These congeners also
stimulate insulin release from a human beta-receptor cell line, reduce synthesis of the neurotransmitter
dopamine in neurons, and activate neurophils to produce reactive oxygen species /17/.
Not only do different PCB congeners have unique sites and mechanisms of action but also their
metabolites have biological activity and are persistent in living organisms for a long period of time /1819/.
The endocrine disruptive actions of PCBs can be a result of the parent PCB, the enzymes induced by the
parent PCB, or due to an action of metabolites /20/. The coplanar and AhR-activating PCBs, for example, are
anti-estrogenic because they induce P450s 1A1 and 1B1, which degrade estrogen /21/. In contrast, most other
PCBs, and especially their hydroxylatedmetab-olites, are estrogenic. Because the great majority of PCBs are
not AhR activators, the net activity of most PCB mixtures is to mimic the actions of estrogen. This variation
in action found in different PCB congeners is particularly important in developing an understanding of the
relation between exposure to PCBs and the risk of breast cancer. Estrogen is by far the best-documented risk
factor for breast cancer /22/. Thus, exposure to AhR-activating PCBs might be expected to be protective
against breast cancer based only on this action, whereas exposure to the estrogenic congeners might be
expected to promote risk.
Because PCBs are found in the lipid fraction, blood levels are reported either as wet weight
concentrations or with the resultslipid-adjusted. Studies from the U.S. Centers for Disease Control and
Prevention (CDC) have shown no difference between fasting and non-fasting results, providing that a lipid
adjustment is made /23/. Nevertheless, lipid adjustment also has some problems in that PCBs can alter lipid
metabolism /24/ and because the method has been reported to be highly prone to bias /25/. The various ways
in which PCB levels are reported and the varying numbers of PCB congeners determined by various
laboratories often make it difficult to compare across studies.
The half-life of PCBs in the human body is long, but varies with the congener, in that the lower
chlorinated congeners are more rapidly metabolized and many of the highly chlorinated congeners persist for
many years. The rate of removal is also a function of body burden. Wolff et al. /26/ reported a half-life of 35
years for individuals with high serum PCBs, but 1317 years for those having lower values.
67
The U.S. Agency for Toxic Substances and Disease Registry (ATSDR) /27/ states that PCB levels in
persons without unusual exposure range from 0.9 to 1.5 ppb wet weight and summarizes information from
many studies showing that PCB levels increase with age. A recent study of older pregnant women (mean age
39 y) in Western Canada reported a mean wet weight concentration of 0.78 ppb, with the range being 0.43 to
3.34 ppb /28/. Thus, it is likely that the ATSDR values overestimate the background serum concentrations of
PCBs in persons without some unusual source of exposure and in persons less than 50 years of age. At older
ages, the ATSDR values are probably accurate.
Routes of Exposure
Ingestion has usually been assumed the primary route of exposure to PCBs /27/. These compounds are fat
soluble and bioaccumulative. The tissues of fish taken from contaminated fresh water are particularly high in
PCBs /27/, although farmed fish like salmon are also an important route of exposure /29/. Everyone in
developed countries has a certain amount of PCBs in their bodies, primarily from contamination of the food
supply /30/. Inhalation and dermal absorption are also possible routes of exposure. Animal studies have
shown that the inhalation of vapor-phase PCBs is an important route of exposure and that inhaled vapor phase
PCBs can bioaccumulate and cause pathological changes /31/. Vapor-phase PCBs are significantly elevated
near PCB-contaminated hazardous waste sites /3233/. The results of our investigations provide evidence that
inhalation is also an important route of exposure in humans /4/ and a cause of various health effects among
individuals who live near such contaminated sites /3436/. Liebl et al. /37/ also demonstrated the
accumulation of lower-chlorinated PCB congeners in humans, a result of breathing contaminated indoor air.
The lower-chlorinated congeners are more volatile but more easily metabolized by the body. Thus, the
appearance of transient congeners in human blood reflects relatively recent exposure, independent of the route
of exposure, but is characteristic of inhalation. Because of the relatively rapid disappearance of the more
volatile congeners, the relative magnitude of exposure by inhalation may be underestimated upon serum PCB
measurement.
oxygen species and the induction of a variety of genes /39/. Chronic exposure to PCBs results in
chromosomal aberrations /40/, an action probably mediated primarily by metabolites of the PCBs, although
they are generally not viewed as being cancer initiators /41/. Nevertheless, evidence from one study indicates
that dioxin-like congeners and dioxin itself produce oxidative DNA damage /42/. In a study investigating the
induction of hepatic tumors in rats, van der Plas et al. /43/ concluded that the majority of the tumor
promotion potential of PCB mixtures resides in the non-dioxin-like fraction, which is not taken into account
in the toxic equivalency factor (TEF) approach for risk assessment of PCBs.
Therefore, one cannot adequately assess cancer risk from PCB exposure by using TEFs. The situation is
further complicated by evidence for interactions, both antagonistic and synergistic, in certain combinations of
PCB congeners, with or without dioxins /4445/. By virtue of these actions, one would expect that they
would result in an increased risk of every kind of cancer /46/.
Studies of cancer in human populations are primarily of two major typesoccupational studies of
workers and case-control studies of individuals with a specific kind of cancer. Polychlorinated biphenyls are
classified as probable human carcinogens by the World Health Organization, based on proof in animals and
results consistent with this conclusion in humans, whereas dioxins are classified as known human
carcinogens. A major reason that the evidence for PCB carcinogenicity does not reach the level of proof in
humans is that PCBs accumulate with many other fat-soluble contaminants, therefore, it is impossible to be
100% confident that the cancer is caused by PCBs and not by DDT or other fat-soluble chemicals.
Nevertheless, it is almost certain that a substance will be carcinogenic in humans if that chemical is
carcinogenic in animals whoshare a majority of our genes. Therefore, when evaluating the impact of PCB
exposure on human health, discounting animal carcinogenic studies has no merit.
Occupational studies are limited in power, especially because (a) the number of employees is usually
small and (b) cancer is a relatively rare disease, for which the incidence in a small population will not be
large, even if the risk is elevated. A number of occupational studies have reported increased numbers of
various PCB-associated cancersliver, gall bladder, biliary tract, leukemia, gastrointestinal, skin (especially
malignant melanoma), lymphoma, lung, pancreatic /27, 47/, but these studiesdo not report PCB levels and are
not of great use in defining risk. The other problem is that not all occupational studies show elevations of the
same type of cancer, probably because of the size of the groups studied. In addition, many occupational
studies are going to show what is known as the healthy worker effect, which is that employed persons are in
general healthier than those who are unemployed. Such limitations are clearly shown in the studies conducted
by Kimbrough et al. /4849/. Although widely reported to be the largest occupational cohort studied, the
study of over 7,000 employees at two General Electric capacitor plants in New York included everyone who
had worked at the plant for a period as short as 90 days, including secretaries and workers that we would not
expect to be exposed. These studiesfocused on mortality,notincidence, and suffered from exposure
misclassification, failure to account for latency of cancer development, and poor statistical power /5051/.
Brain cancer. Sinks et al. /52/ reported an elevated incidence of brain cancer in workers in an Indiana
capacitor-manufacturing plant. Ruder et al./53/ recently published a follow-up study of the same population
and found a SMR of 1.91 (1.0-3.3) and a significant dose-response trend (p = 0.016). The highest SMRs were
found among workers in the highest tertile of PCB exposure.
Breast cancer. Many small clinic-based breast cancer studies have been published, some of which
suggest a dose-dependent relation to total PCB exposure, but most large retrospective and nested
epidemiologic studies have not demonstrated a relation between elevated total PCB levels and breast cancer.
These have been reviewed by Moysich, et al. /54/. As discussed above, such conflicting observations may be
a result of the different endocrine actions of different PCB congeners. Nevertheless, some evidence has been
found for a relation between PCB exposure and risk of breast cancer in individuals with exposure to certain
PCB congeners and in persons from certain racial or genetically susceptible subgroups.
69
PCBs are potent inducers of the enzyme cytochrome CYP1A1, which has several variant genotypes. In a
retrospective case-control study, Moysich et al. /55/ looked at the risk of breast cancer in relation to serum
PCB levels in relation to the specific genetic polymorphism of P450 1A1, which codes for a isoleucine to
valine substitution. Overall, postmenopausal womenhaving a higher body burden of PCBs were not at greater
risk ofbreast cancer than were women with lower levels. When stratified by genotype, however, among
women with serum PCB levels above the median of the distribution in the control group, increased risk of
breast cancer was associated with the presence of at least one valine allele (either CYP1A1 Ile:Val or Val:Val)
(OR 2.93; 95% CI 1.177.36) when compared with women who were homozygous for the isoleucine alleles
(CYP1A1 Ile:Ile). Women with lower PCB levels and the CYP1A1 Ile:Val or Val:Valgenotypes werenot at
greater risk for breast cancer, nor were women with elevated PCB levelsandCYP1A1 Ile:Ile.
Millikan et al. /56/ studied plasma DDE (1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene) and PCB levels in
relation to breast cancer in a population-based, case-control study of African-American and Caucasian
women. The ORs for the highest to lowest third for total PCBs were 1.74 (95% CI: 1.003.01) for AfricanAmerican women and 1.03 (95% CI: 0.681.56) for Caucasian women. For obese African-American women,
the OR was 4.92 (95% CI: 1.6314.83). These observations are consistent with the presence of
subpopulations that are genetically at greater risk of developing breast cancer following exposure to PCBs.
Aronson et al. /57/ examined breast tissue from 217 hospital cases of cancer and 213 controls. The
authors found no relation with total PCBs but significant correlations with PCBs 105 and 118. The ORs for
these two congeners increased linearly across categories (p for trend < 0.01). The elevated risk associated
with PCBs 105 and 118 were higher among premenopausal women, whereas for postmenopausal women the
risks were elevated with PCBs 170 and 180.
Demers et al. /58/ studied 315 cases of breast cancer and 523 hospital and population controls. The
authors reported significant elevations of three individual PCB congeners, PCB 99, 118, and 156, with
significant associations between breast cancer risk and PCB 118 (OR = 1.60, 95% CI: 1.012.53) or PCB 156
(OR = 1.80, 95% CI: 1.112.94). The group also found a significant relation with the sum of mono-orthoPCBs 105, 118, and 156 (OR = 2.02; 95% CI: 1.24, 3.28, 1stvs 4th quartile). These observations suggest that
certain congeners or combinations of congeners cause a dose-dependent increase in the risk of breast cancer,
whereas others do not. Although less information is available on the PCB-induced risk of other estrogendependent cancers, what is true for breast cancer is likely true for other types of cancers.
Holford et al. /59/ analyzed nine congeners in a hospital-based case-control study and reported that
although no relation between total PCBs and breast cancer risk could be established, an anti-estrogenic and
dioxin-like congener, PCB 156, had a protective effect, whereas two phenobarbital-like congeners PCBs 180
and 183 had adverse effects. The latter report indicates the importance of obtaining congener-specific
information.
Gastrointestinal cancers.Mallin et al. /60/ reported a mortality study of workers employed at a capacitor
manufacturing plant using PCBs and found an elevated incidence of intestinal cancer in women employed for 5
or more years (SMR 2.2) and an elevated risk of stomach cancer in men (SMR 2.2). Howsam et al. /61/ reported
an elevated risk of colorectal cancer with increased levels of the mono-ortho congeners PCBs 28 and 118. In the
top tertile, the OR was 2.94 (95% CI: 1.396.20). Gastrointestinal cancers have been reported to be significantly
elevated in some /62/ but not all occupational studies of PCB exposure. Hoque et al. /63/ reported a dosedependent increase in gastrointestinal cancers in a Michigan cohort exposed to polybrominated biphenyls
(PBBs). For PBB levels of 420, 2150, and > 50 ppb, the respective ORs were 8.23 (95% CI: 1.2753.3), 12.3
(95% CI: 0.80191), and 22.9 (95% CI: 1.34392).
Liver/biliary cancers.Liver and biliary cancers are some of the most common neoplasms induced in
experimental animals exposed to PCBs /6465/. Such tumors have also been reported in human occupational
studies /6667/. In the capacitor study of Mallin et al. /60/, an SMR of 6.2 was found for liver/biliary cancer
70
Hoque et al. /63/ studied various cancers in 187 persons exposed to PBBs, which would be expected to
have the same effect as PCBs, and 696 unexposed controls. The authors found no overall increased risk of
cancer related to serum PBB level, but did find a PBB-related dose-dependent elevated risk for both
lymphoma and digestive system cancers. For lymphoma, taking a PBB level of < 3 ppb as control, the ORs
were 3.85, 19.6, and 48.9 for the respective PBB levels of 420, 2150, and > 50 ppb, all of which are highly
significant.
Pancreatic cancer.Yassi et al. /78/ reported a significant elevation of pancreatic cancer mortality in
workers in a transformer manufacturing plant using transformer fluids, some containing PCBs (SMRs ranging
from 2.927.64). Studying a group of 108 cases and 82 controls, Hoppin et al. /79/ have shown that exposure
to PCBs results in a significant dose-dependent increased risk of pancreatic cancer (p for trend < 0.0.001).
Total PCBs were measured in serum and reported as lipid-adjusted values. Taking lipid-adjusted PCB levels
in serum below 185 ppb as reference, a 1.3-fold risk (95% CI: 0.62.8) was found for levels between 185
360 ppb, and a 4.2-fold increased risk for levels over 360 ppb (95% CI: 1.99.4). Not only is the last number
statistically significant but also a p for trend and a continuous (OR = 1.003 for each ppb increase in
concentration) variable are significant as well. Significantly elevated ORs were reported for the highest
tertiles of PCB congeners 153 (OR 3.0, 95% CI: 1.46.6) and PCB 180 (OR = 8.4, 95% CI: 3.421).
In a study of 51 cases of pancreatic cancer by Porta et al. /80/, cases with wild-type K-ras gene1 (n=17) were
frequency matched for age and sex to cases having a K-ras mutation (n=34, case-case study). The authors found
that cases having a K-rasmutation had higher concentrations of PCBs 153, 180, and 138 (the only congeners
measured). When compared with 26 hospital controls (case-control comparison), the total concentrations of the
three PCBs were higher in the 51 cases than in the 26 controls, but the differences were significant only for PCB
180. The 51 cases were more than four times more likely than the 26 controls to be in the upper tertile of PCB
180 (OR = 4.6, 95% CI: 1.119.0, p for trend = 0.037). The OR for the 34 K-ras-mutated cases was 7.4 (95%
CI: 1.634.4, p for trend = 0.012). Considered alone, lipid adjusted, and taking non-detected values as OR = 1.0,
in tertiles, the ORs for PCB 138 increased from 1.0 to 2.9 (95% CI: 0.517.2) to 6.9 (95% CI: 1.141.5), with a
p for trend of 0.034. For PCB 153, the ORs ranged from 1.0 to 1.8 (95% CI: 0.47.6) to 7.2 (95% CI: 1.145.6);
p for trend = 0.035. For PCB 180, the ORs ranged from 1.0 to 2.8 (95% CI: 0.614.3) to 6.3 (95% CI: 1.0
38.8); p for trend = 0.028.
Prostate cancer. For three specific groups of PCB congeners, Ritchie et al. /8182/ demonstrated a dosedependent increased risk of prostate cancer as a function of serum PCB concentration. Interestingly, the authors
did not find an increased risk with dioxin-like congeners, but did find a dose-dependent increased risk of up to
more than 2 orders of magnitude for moderately chlorinated congeners and transient and persistent
phenolbarbital-like inducers. This study is one of the clearest demon-strations that not only dioxin-like PCB
congeners can increase the risk of cancer. In an occupational nested case-control study, Charles et al. /83/
reported an OR of 1.47 (95% CI: 0.972.24) for serum PCB levels and prostate cancer mortality after
adjustment for suspected confounding factors.
Thyroid cancer. Thyroid cancer is commonly induced in animals upon exposure to PCBs /65, 84/. The
occupational study of capacitor workers conducted by Mallin et al. /60/ reported a significant SMR of 15.2
for thyroid cancer in men.
Family of retrovirus-associated (ras) DNA sequencesisolated from Harvey (H-ras, Ha-ras, rasH) and Kirsten (K-ras, Ki-ras, rasK)
murine sarcoma viruses. Sequences corresponding to both H-ras and K-ras genes have been detected in human, avian, murine, and
non-vertebrate genomes.
72
Recurrent Infections
Suppression of the immune system is an important factor because individuals with sub-normal immunity
show increased susceptibility to infections and to cancer. As exposure to PCBs can suppress both the
antibody (immunoglobulins) and the cellular immune response, frequent infections can be a direct result of
PCB exposure. Until recently, the ATSDR considered immune suppres-sion to be the biologic effect that
occurred at the lowest PCB concentration, although they now believe that effects on neurobehavior occur at
even lower doses.
Human studies have clearly shown that persons exposed to PCBs have a greater incidence of all kinds of
infections. One year after the event,Lu and Wu /85/ studied patients in Taiwan who were exposed to PCBcontaminated rice oil in 1979. As compared with controls, exposed patients had more infections, especially of
the respiratory tract and skin. The initial PCB levels in the patients varied between 3 to 1156 ppb (mean = 89
ppb). When tested later, a subgroup showed elevated immunoglobulin classes IgA and IgM, a reduced
delayed-type response to skin test antigens, and a reduced number of several classes of white blood cells
involved in the immune response. This study does not report PCB levels in these subjects, but the differences
are significant relative to unexposed controls.
Weisglas-Kuperus et al. /86/ showed that at 3 months of age,Dutchchildren exposed to a mixture of PCBs
and dioxins had lower levels of monocytes and granulocytesthan did less exposed children. As these white
blood cellsscavenge and kill microbes, an abnormally low concentration reduces the bodys resistance to
many infections. In a later study /87/ of 207 Dutch mother-infant pairs, the same group measured the sum of
PCB congeners 118, 138, 153, and 180 in the mothers serum, in cord blood, and in breast milk. The authors
state that these four congeners constituted 46% of the total PCBs. The range of PCB levels in children at 42
months of age was from 0.08 to 5.90 ppb, with the average being well below 1 ppb. Adjusted for
confounders, higher PCB levels were associated with a higher incidence of recurrent middle-ear infections
(elevated 3-fold) and of chicken pox (elevated 7.6-fold), but a lower prevalence of asthma. The children with
higher PCB levels had more coughing, chest congestion, and phlegm. The authors conclude that the higher
the PCB level, the greater the frequency and severity of infections,but the lower the frequency of allergic
diseases.
These children were followed up until school age and found to have a persistently higher prevalence of
ear infections /88/. Dewailly et al. /89/ found that one-year-old infants fed milk contaminated with PCBs had
a 20-fold higher incidence of infectious diseases, such as measles, meningitis, and middle ear infections than
did children with less exposure.
Immunoglobulins(Ig) produced by lympho-cytesare good markers for the integrity of the immune system.
In a study of Belgian adolescents, Van Den Heuvel et al. /90/ found that as levels of three PCB congeners and
total dioxin equivalents (TEQs) increased, serum IgA levels increased and IgG and IgE levels decreased.
We recently studied the hospitalization rates for infectious diseases in relation to residence near PCBcontaminated sites.Kudyakov et al. /35/ found that the hospitalization rates for acute and chronic infectious
respiratory diseases were significantly elevated in persons living in PCB-contaminated zip codes in upstate
New Yorkwhencompared with clean zip codes without hazardous waste sites or with zip codes having
hazardous waste sites containing other kinds of wastes. As a control for other well-documented confounders,
the authors report that the family income of individuals living along the Hudson River is greater than that of
other upstate residents and, based on the Behavioral Risk Factor Surveillance System, such persons smoke
less, exercise more, and eat more fruits and vegetables than do other New Yorkers. Yet the Hudson River
residents show significantly higher rates of hospitalization for chronic respiratory infections. We attribute the
elevated rates to inhalation exposure to PCBs coming from the Hudson River, resulting in suppression of
immune function.
73
Neurobehavioral Effects
In utero exposure to PCBs has been linked to adverse effects on intellectual function in infants and young
children. Jacobson and Jacobson /91/ assessed whether such effects persist through school age in children
born to women who during pregnancy had eaten fish contaminated with PCBs. The authors followed 212
children from birth to 11 years of age. PCB levels were determined in maternal serum, in breast milk, in the
umbilical cord blood at delivery, and in the childrens blood 11 years later. The authors then administered a
battery of IQ and achievement tests. The average maternal serum level at the time of the childs birth was 6
ppb, whereas the childs was 1 ppb at age 11 years (not lipid adjusted). The breast milk on average contained
841 ppb of PCBs, lipid adjusted. The results show a significant reduction in full-scale IQ in children of
mothers whose breast milk contained PCBs at a concentration of 1,250 ppb (lipid adjusted) or greater and
poorer performance on reading mastery in children whose mothers had 1,000 ppb or greater PCBs in breast
milk. The authors conclude that perinatal exposure to PCBs causes an irreversible decrement of IQ.
Chen et al. /92/ reported on the cognitive development of children born to mothers who had eaten PCBcontaminated rice oil and dibenzofuransin Taiwan in 1976. These authors studied 118 children born to
exposed mothers (some born years after the poisoning) and 118 matched controls. The exposed children
scored approximately 5 points lower on the Wechsler Intelligence Scale for Children. The children were
followed through ages 4, 5, 6, and 7 years, but no obvious improvement occurred with time. This study did
not obtain PCB levels but reports that after the exposure, the average levels were 49.3 ppb in adults (range
2.0456 ppb) in the exposed population, whereas the mean PCB level was 9.8 ppb in 92 Taiwanese blood
donors (not used as controls in this study, but perhaps reflective of the population).
Lonky et al. /93/ attempted to replicate the studies carried out by Jacobson through studying children born
to mothers who had eaten PCB-contaminatedLakeOntario fish. This paper does not give PCB values (they
studied only cord blood, but the data appear in a different paper), but categorizes mothers either as no fish,
low fish, or high fish. Each group comprised over 150 mother-infant pairs. Using the Neonatal
Behavioral Assessment Scale to study neurologic development over the first 48 h of life, the investigators
found that infants in the high fish category showed more abnormal reflexes, greater responses to stress, and
less habituation to repeated stimuli than did no fish and low fish babies. The results are consistent with
known effects of PCBs on the brain before birth.
Rogan et al. /94/ studied 912 children in North Carolina born to mothers with no known special exposure
to PCBs and performed the Brazelton Neonatal Behavioral Assessment Scales (psycho-logic and neurologic
tests designed for use in newborn infants) in relation to the PCB levels in breast milk. Children who were
more exposed showed low muscle tone and depressed reflexes. Relating the breast milk PCB levels reported
in this study to those of other studies is hard, but the Rogan study is important because it shows that levels
that are present in the general population (about 1980 ppb) alter the nervous system performance of children.
Walkowiak et al. /95/ reported on 171 mother-infant pairs from Germany who were recruited into the
study between 1993 and 1995. The children were examined repeatedly for psycho-development at 7, 18, 30,
and 42 months of age. Prenatal and perinatal PCB levels were determined in cord blood and breast milk and
in the childrens serum at 42 months of age. Only three PCB congeners were measured at levels representing
European ambient background exposure. Prenatal exposure to PCBs had a negative effect on both mental and
motor development in children at all ages. The results show that even at background exposure levels, more is
worse, but because the investigators measured only three congeners, determining the relative risk is difficult.
Yu et al. /96/ studied behavior in 118 children born to the Taiwanese mothers who were exposed to PCBs
by consuming rice oil and matched controls. Consistent and significant differences were found on the Chinese
version of Rutters Child Behavior Scale A measuring emotional and behavioral disorders on health, habit,
and behavior. The exposed children were examined annually from 1985 to 1991 and performed more poorly
74
than controls on all three components, which persisted as the children aged.
In an assessment of impact of PCB exposure during adulthood on intellectual functioning, Schantz et al.
/97/ studied memory function in 572 49- to 86-year-old adults who eat a great amount of contaminated Great
Lakes fish and 419 people who do not. Three memory tests were used, as well as several visual tests, and
serum PCBs and a number of other contaminants were measured. The PCB levels were divided into four
groups: ND4.6 ppb, 4.77.8 ppb, 7.913.8 ppb, and 13.975.0 ppb. Inall three memory tests, performance
decreased as the PCB dose increased, butPCB exposure had no effect on the visual tests. No other
contaminant studied was associated with poor performance on memory tests. This study is important because
it is the only one clearly showing that even adults can suffer from a specific loss of IQ and memory upon
PCB exposure.
The general conclusion is that the higher the childs exposure to PCBs in early life, the lower the IQ and
the more the child exhibits anti-social behavior, depression, and attention deficit hyper-activity disorder-type
symptoms. Theseeffectsare found over the full range of IQ, however, and even bright kids would have been
brighter had they not been exposed. In adults exposed to PCBs, a decre-ment in IQ is paralleled by a
reduction of memory.
Hypothyroidism
Thyroid hormones have some structural similarity to PCBs but have iodine rather than chlorine
substitutions on the two phenyl rings. A number of animal studies clearly show that PCBs interfere with
thyroid hormone at multiple sites /98/.
Koopman-Essenboom et al. /99/ studied thyroid function in mother-infant pairs in the Dutch study /88/ and
found that high organochlorine levels in mothers milk correlates with low plasma maternal T3 and T4 levels
and with high thyroid stimulating hormone (TSH, thyrotropin) levels in their infants. Osius et al. /100/
measured thyroid hormones and eight PCB congeners in blood samples obtained from 1,091 second-grade
children living near an incinerator in Germany. One congener (PCB 118) correlated positively with TSH,
whereas five other congeners were negatively correlated with free T3, the active form of thyroid hormone.
Schell et al. /101/ studied PCBs and thyroid hormones in Native American adolescents living near the St.
Lawrence River where the fish are contaminated with PCBs. The mean PCB level was 1.82 ppb and the
maximum level 4.75 ppb. Schell found a statistically significant positive and dose-dependent relation between
total PCBs and TSH levels and a significant negative relation with both free and total thyroxine (T4). A
negative relation to T3 (triiodothyronine) was found, but not at a level that was significant. This study shows
clearly that PCBs at levels common in the population reduce thyroid function. Wang et al. /102/ reported
reduced levels of free T4 FT4 TSH (based on the normal hypothalamicpituitary axis in which decreased T4
feeds back to the hypothalamus and stimulates the anterior pituitary to secret TSH)with increasing levels of
placental non-orthoPCBsin neonates from the general population.
Taiwan exposed to PCBs in 19781979 and studied by Hsu et al. /109/ in 19992002 were found to have
more abnormal sperm than did controls; the sperm they did have showed a reduced capability to bind and
penetrate oocytes.
For both dioxins /110/ and PCBs /111/, clear evidence shows that the ratio of male to female births is
strikingly reduced following parental exposure. Exposure to these compounds also alters female reproduction.
Both PCB /112/ and PBB /113/ exposure causes an earlier menarche in girls. Cooper et al. /114/ reported that
women show a positive association between PCB levels and increasing menstrual cycle length (p = 0.02). The
exposure of monkeys to dioxins and dioxin-like PCBs results in endometriosis /115117/. Pauwels et al. /118/
showed an elevated incidence of endometriosis in humans with higher dioxin toxic equivalents, and Buck-Louis
et al. /119/ reported that women in the third tertile of anti-estrogenic PCBs have an OR of 3.77 (95% CI: 1.12
12.68) for visually confirmed endometriosis, which is consistent with the monkey evidence.
contaminant. The authors found that the rate of diagnosis of coronary heart disease were 15% higher in
residents who live in zip codes containing or abutting a POPs sites, and that rates of myocardial infarction
were 20% higher in these zip codes. Sergeev and Carpenter also studied a subset of zip codes along the
Hudson River, 200 miles of a National Priority Site highly contaminated with PCBs, where the average
income is higher, smoking rates are lower, residents exercise more frequently and consume fruits and
vegetables more regularly than in the other parts of the state. Despite a higher socio-economic status and
healthier life style, the authors found a 35.8% higher frequency diagnosis of coronary heart disease and a
39.1% more frequent diagnosis of myocardial infarction in this population.
Hypertension
Kreiss et al. /126/ reported that residents of Triana, Alabama having high serum PCB concentrations from
eating local contaminated fish showed a significantly higher incidence of high blood pressure than those with
lower PCB levels. The increase in blood pressure was independent of age, sex, body mass index, and social
class. In a population of 458 persons with an average serum PCB concentration of 17.2 ppb, the rate of
borderline or definite hypertension was 30% higher than expected from national rates. The relation with PCB
concentration was highly significant for diastolic blood pressure and of borderline significance for systolic
blood pressure. Excess hypertension has been reported in several occupational studies of PCB exposure/135
137/.
Conclusions
Polychlorinated biphenyls alter the functioning of many different organ systems in both animals and
humans and are risk factors for a large number of human diseases. Although levels in the environment and in
populations are declining since their manufacture and use has been reduced, these substances are persistent
and remain in the environment and in the human body. It is imperative to continue to reduce human exposure
to these dangerous compounds.
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7127), BP 74, 29682 RoscoffCedex, France
Received July 18, 2001
78
To assess human health risk from environmental chemicals, we have studied the effect on cell cycle regulation of the widely
used glyphosate-containing pesticide Roundup. As a model system we have used sea urchin embryonic first divisions
following fertilization, which are appropriate for the study of universal cell cycle regulation without interference with
transcription. We show that 0.8% Roundup (containing 8 mM glyphosate) induces a delay in the kinetic of the first cell
cleavage of sea urchin embryos. The delay is dependent on the concentration of Roundup. The delay in the cell cycle could be
induced using increasing glyphosate concentrations (1-10 mM) in the presence of a subthreshold concentration of Roundup
0.2%, while glyphosate alone was ineffective, thus indicating synergy between glyphosate and Roundup formulation products.
The effect of Roundup was not lethal and involved a delay in entry into M-phase of the cell cycle, as judged cytologically.
Since CDK1/ cyclin B regulates universally the M-phase of the cell cycle, we analyzed CDK1/cyclin B activation during the
first division of early development. Roundup delayed the activation of CDK1/cyclin B in vivo. Roundup inhibited also the
global protein synthetic rate without preventing the accumulation of cyclin B. In summary, Roundup affects cell cycle
regulation by delaying activation of the CDK1/cyclin B complex, by synergic effect of glyphosate and formulation products.
Considering the universality among species of the CDK1/cyclin B regulator, our results question the safety of glyphosate and
Roundup on human health.
98 29 23 46. Fax: 33 (0) 2 98 29 23 06. E-mail: belle@sb-roscoff.fr.
Introduction
Widespread use of pesticides leads to chronic
exposure to a combination of various products.
Despite the obvious benefits of pesticides, their
extensive use has posed problems for both
environment and human health. Among the human
health concerns, exposure to several groups of
pesticides has been associated with various cancers (1,
2) or reproductive and developmental disor-ders (3).
Therefore, searches for mechanisms by which
pesticides could interfere with cell cycle or
developmental processes are of great interest.
The cell cycle is the universal process by which
cells reproduce and underlies the growth and
development of all living organisms. Controls operate
during the cell cycle to regulate the onset of DNA
replication and segregation of the replicated
chromosomes. Surveillance mechanisms, called
checkpoint (4) pathways, ensure the proper order and
correct execution of cell cycle events. The molecular
basis of these controls is highly conserved from single
unicellular eucaryotes to complex metazoans such as
humans (for review, see refs 5-7). Human cancers are
associated with defects in the control of cell cycle
transitions (8). Errors in the choreography of the
process of mitosis and cytokinesis can lead to genetic
instability fostering cell death or disease. Failures of
these checkpoints allow cells to divide when DNA is
incorrectly replicated, or when chromosomes are
incor-
10.1021/tx015543g CCC: $22.00 2002 American Chemical Society Published on Web 02/22/2002
79
Figure 1. Kinetics of the first division of early development.Sea urchin gametes were fertilized and the embryos transferred 10 min postfertilization
into fresh seawater (full circles) or into seawater containing 0.8% Roundup (open circles). One hundred embryos were scored for the first cleavage by
phase contrast microscopy as a function of time after fertilization. Insert: nuclear division shown at 150 min following fertilization scored by
fluorescence microscopy observation of control- and Roundup-treated embryos stained with Hoechst dye. Bar: 90 M.
80
suc1
presence of 50 L p13
-Sepharose beads. After three washes, the activity of the bound kinase was determined under the standard conditions
reported above.
Determination of Protein Synthesis Rate in Vivo.
35
Batches of embryos (1 mL) were incubated in the presence of 10 Ci/mL [ S]-L-methionine. After 5 min pulse incubations, embryos were
35
rapidly pelleted and frozen. [ S]Methionine incorporation into proteins was measured on duplicate aliquots of the embryo extracts after 10%
TCA precipitation on Whatman 3M filters and counting in the presence of OptiphaseSupermix scintillation liquid.
Analysis of Cyclin B by Immunoblot. Cyclin B detectionwas performed on whole extracts from embryos taken every 15 min after
fertilization. Pellets from 200 L of a 5% egg suspension were boiled for 3 min in 200 L of electrophoresis sample buffer (17). After clearing
by centrifugation, 10 L of the embryos extract were resolved on 12% acrylamide gel electrophoresis (17), transferred on nitrocellulose (18),
stained by Ponceau Red and processed for western blotting using anti-cyclin B antibody (13) detected with goat anti-rabbit Ig G (H+L)-AP
conjugate (Bio-Rad) using the NBT-BCIP reagent (Fulka).
Results
Effect of Roundup on the First Cell Cycle. Seaurchin early development was used to monitor cellular dysfunction induced
by Roundup. Several steps of early development were affected by the presence of Roundup in the incubation medium.
Abnormality in the first cell cycle of early development was observed and further analyzed.
The presence of Roundup at 0.8% (containing 8 mM glyphosate) applied at 10 min postfertilization affected the first cell
cycle as judged by a delay in the occurrence of the first cleavage. In a typical experiment (Figure 1), the cleavage occurred 120
min postfertilization in the earliest embryos and had occurred in 100% of the embryos at 210 min, whereas cleavage started at
165 min postfertilization in the earliest pesticide-treated embryos and had occurred in all pesticide-treated embryos at 300 min.
Pesticide-treated embryos were undistinguishable from the control foster embryos at the cytological level after Hoechst
staining (see Figure 4). The insert of Figure 1
328 Chem. Res. Toxicol., Vol. 15, No. 3, 2002
Figure 2. Dose-response effect of Roundup on the first divisionof early development. Sea urchin gametes were fertilized and the embryos transferred
10 min postfertilization into fresh seawater or into seawater containing Roundup at various indicated concentrations. The time at which 50% of the
embryos cleaved was determined by observation of the embryos by phase contrast microscopy and compared for untreated versus treated embryos in
nine different experiments. Vertical bars represent the standard deviation for the nine independent experiments.
81
Figure 3. Synergic effects of Roundup and glyphosate on thefirst division of early development. Sea urchin gametes were fertilized and the embryos
transferred 10 min postfertilization into fresh seawater containing increasing concentrations of glyphosate, together with 0.2% Roundup which already
contains 2 mM glyphosate (open circles) or without Roundup (full circles). The time at which 50% of the embryos cleaved was determined by phase
contrast microscopy and compared in control-versus treated-embryos. The figure is representative of three indepen-dent experiments.
illustrates embryos at 150 min postfertilization. Embryos remained healthy as judged by further development of almost all
treated embryos through all the cleavage stage. When the pesticide-treated embryos were incubated up to 6 h and then
transferred in pesticide-free seawater, their development proceeded as for the control embryos reaching the prisme stage 2 days
postfertilization, thus indicating that 6 h incubation in 0.8% Roundup was not lethal and did not provoke early developmental
irrevers-ible damage.
The effect of various concentrations of Roundup on the first cell cycle delay was analyzed as the time at which 50% of the
embryos had cleaved. Kinetics of first cleavage among different batches of embryos show inherent bio-logical variability.
Comparison could be performed using the ratio between the times necessary to reach 50% of first mitotic division for the
control embryos versus that for the foster treated-embryos. The delay in the first cleavage kinetic was reproducibly observed
and was proportional to the concentration of Roundup (Figure 2). In all experiments, the dose-response effect of Roundup
occurred in a narrow concentration window (see FigureMarc et al.2). A concentration of Roundup of 0.68% (containing 6.8
mM glyphosate) was estimated to produce half the maximal delay effect. When Roundup concentration exceeded 1%
(containing 10 mM glyphosate), develop-ment was generally arrested at the first cell cycle stage.
The minimal exposure time required for cell cycle dysfunction was examined by scoring first cleavage under various
exposure conditions. In a set of incubations, Roundup was added to seawater at different times after fertilization and
maintained continuously. A significant delay in first cleavage was observed when the pesticide was added at 10, 30, or 60 min
postfertilization. Added after 90 min, Roundup was no longer able to delay the first mitotic division. In a second set of
incubations, Roundup was transiently added to the medium for different time intervals. A 30 min pulse exposure, applied from
10, 30, or 60 min postfertilization, did not delay the first cleavage. On the other hand, a 60 min exposure to Roundup, applied
from 10 or 30 min postfertilization, consistently delayed the first mitotic division. Therefore, dysfunctions in the first cell cycle
occur if 0.8% Roundup applied to embryos for at least 60 min is starting within the first hour following fertilization.
Effect of Glyphosate on the First Cell Cycle. Thecommercial Roundup contains glyphosate, the active product as
pesticide, associated with a combination of formulation products. The effect of pure glyphosate on the first cleavage was
investigated. At concentrations of glyphosate ranging from 1 mM up to 20 mM (correspond-ing to the amount present in
Roundup 0.1-2%), applied under the same experimental conditions as Roundup, pure glyphosate had no effect on the first
mitotic cell division. Therefore, we conclude that either the formula-tion products of Roundup are directly responsible for the
cell cycle dysfunction or glyphosate and formulation products act in synergy.
Possible synergy between the formulation products and glyphosate was assayed by analyzing the effects of various
concentrations of glyphosate on the first cell cycle, in the absence or in combination with subthreshold doses of Roundup.
Increasing concentrations of glyphosate were effective in dysregulating the first cell cycle in the presence of 0.2% Roundup,
whereas 0.2% Roundup alone was inefficient (Figure 3). As observed for the effect of Roundup alone, the combined effect of
glyphosate and Roundup occurred in a narrow concentration range. We conclude that glyphosate and formulation products
present in Roundup exert synergic effects that delay the first cell cycle of early development.
Cellular Analysis of Roundup-Induced Cell Cycle Dysfunction. The effects of Roundup on first cell cyclewere analyzed
at a cellular level. Fertilized embryos were treated with 0.8% Roundup (containing 8 mM glyphosate) at 10 min
postfertilization. At various times, batches of embryos were stained with Hoechst dye and observed by fluorescence
microscopy. Chromatin morphology during all phases of the first cell cycle was comparable in the Roundup-treated and control
embryos (Figure 4). How-ever, stages of chromatin evolution during the cell cycle were delayed in the treated embryos
compared to control embryos (Figure 4). Careful time course observations from fertilization to cytokinesis were performed. No
delay was observed in the Roundup-treated embryos during the first 30 min, from fertilization to the close association of the
two pronuclei. The period between association of the pronuclei up to their fusion, in which the male pronucleus
Roundup-Induced Cell Cycle Dysfunction
82
Figure 4. Changes in chromatin morphology during early development.Sea urchin gametes were fertilized and the embryostransferred 10 min
postfertilization into fresh seawater (control: upper panel) or into seawater containing 0.8% Roundup (Roundup: lower panel). Chromatin state was
observed by fluorescence microscopy of Hoechst stained embryos as a function of time after fertilization. Left, an unfertilized egg. Bar: 20 M.
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Figure 5. Kinetics of CDK1/cyclin B activation during earlydevelopment.Sea urchin gametes were fertilized and the em-bryos transferred
10 min postfertilization into fresh seawater (filled circles), into seawater containing 0.8% Roundup (open circles) or into seawater containing
0.2 mM emetine (open triangles). At the indicated times postfertilization, CDK1/cyclin B activation state was determined from the
measurement of its H1 kinase activity as indicated under material and methods. In this experiment, first cleavage occurs around 170 min for
control embryos and around 240 min for 50% of the Roundup-treated embryos. The figure is representative of four indepen-dent
experiments.
undergoes decondensation, went on for 20 min in control embryos and for 50 min in the Roundup-treated embryos.
The phase of zygotic chromatic condensation lasted around 50 min in both Roundup-treated and control embryos. The
characteristic mitotic stages, metaphase to cytokinesis, occurred with comparable time lags for all embryos.
Thus, Roundup induces a delay in the onset of M-phase of the cell cycle without affecting significantly the kinetic of
mitotic progression itself or the period of cytokinesis.
Effects of Roundup on H1 Kinase Activation in Vivo. The complex CDK1/cyclin B controls entry intoM-phase
(for review, see refs 5-7). We first tested for a direct effect of Roundup on the activity of the protein kinase. Neither
Roundup nor glyphosate inhibited the activity of purified CDK1/cyclin B in vitro. The effect of Roundup was therefore
tested on the activation of CDK1/ cyclin B in vivo, determined from H1 kinase activity of extracts prepared at time
intervals after fertilization. Figure 5 shows that control embryos presented charac-teristic peaks of H1 kinase activity at
105 and 195 min postfertilization corresponding respectively to the first
Figure 6. Kinetics of the increase in protein synthesis duringearly development. Sea urchin gametes were fertilized and the embryos
transferred 10 min postfertilization into fresh seawater (filled circles), into seawater containing 0.8% Roundup (open circles) or into seawater
containing 0.2 mM emetine (open triangles). At the indicated times after fertilization, batches of embryos were processed for protein
35
synthetic rate determina-tion, by 5 min pulse incubations in the presence of [ S]-methionine and measurement of the labeled proteins as
indi-cated under material and methods. In this experiment, first cleavage occurs around 170 min for control embryos and around 240 min for
50% of the Roundup-treated embryos. The figure is representative of four independent experiments.
and second cell cycle. The presence of Roundup 0.8% in the incubation medium totally abolished the peaks of H1
kinase activity. Therefore, Roundup acts on the mecha-nism of activation of the complex.
Inhibition of protein synthesis has been reported to delay or block first cell cycle in sea urchin (16, 19, 20). In our
experimental conditions, the protein synthesis inhibitor, emetine, did inhibit the CDK1/cyclin B activa-tion in vivo
(Figure 5). The effect of Roundup on protein synthesis was therefore investigated. In the control embryos, protein
synthesis rate readily increased after sperm addition as well documented (9) and was pre-vented by emetine treatment
(Figure 6). Roundup 0.8% was found to strongly inhibit the increase in protein synthesis associated with fertilization
(Figure 6).
Analysis of Cyclin B during Early Development.
The synthesis of cyclin B is a prerequisite for the activation of CDK1 (5-7). Roundup might have hindered
330 Chem. Res. Toxicol., Vol. 15, No. 3, 2002
Figure 7. Analysis of cyclin B during early development. Seaurchin gametes were fertilized and the embryos transferred 10 min
postfertilization into fresh seawater or into seawater containing 0.8% Roundup. Batches of embryos were processed for immubolotting using
anti-cyclin B antibody as indicated under material and methods, at the indicated time in minutes postfertilization. Roundup-treated embryos
(R) were compared to control (C) and to unfertilized (U) eggs processed in parallel. Molecular weight markers were run on the same
electrophoretic gel and are indicated (kDa) on the left side. Position of Cyclin B is arrowed on the right side. Control embryos underwent the
first division at 90 min postfertilization, cell division of the Roundup-treated embryos was delayed by 60 min in this experiment.
the activation of CDK1/cyclin B through the deficiency in the amount of cyclin B as a result of the general inhibition
of the protein synthesis machinery. We ana-lyzed the level of cyclin B during early development using a polyclonal
antibody directed against sea urchin cyclin B. The antibody recognized a major single band produced postfertilization
at 46 kDa (Figure 7). The protein amount increased and progressively resolved as a doublet 46-49 kDa at M-phase
(Figure 7). The upper band of the doublet then strongly decreases at anaphase (data not shown). The appearance of the
doublet reflected the phosphorylated form of the protein that takes place after the activation of the CDK1/cyclin B
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complex (5-7). In the Roundup-treated embryos, cyclin was synthesized postfertilization (Figure 7). Resolution of the
protein as a doublet was delayed compared to control (Figure 7), reflecting the delay in CDK1 activation. The
synthetic rate of cyclin synthesis was estimated from densitometry of immunoblots. No significant difference was
found between the Roundup-treated and control embryos indi-cating comparable rates in the accumulation of cyclin B
postfertilization.
Discussion
Our results demonstrate that Roundup affects the first cell cycle of early development by impeding activation of the
cell cycle regulator CDK1/cyclin B. Cell cycle is regulated through an all-or-nothing switch of CDK/cyclin activity
after progressive synthesis of the cyclin compo-nent (7). The narrow concentration range of Roundup efficiency (see
Figure 2 and 3) may be related to this all-or-nothing mechanism. The switch is under posttrans-lational control by
protein dephosphorylation/phospho-rylation modifications. The timing of the entry into M-Phase is dependent on the
activation of the mitotic switch that is under the control of the checkpoint pathways (for review, see refs 5-7). Since
CDKs and cell cycle checkpoints are universal from unicellular eucary-otes to human, the molecular target of Roundup
is likely to be of universal meaning. Deregulations of cell cycle checkpoints are directly linked to genomic instability,
which can generate diseases and cancer (see introduc-tion). Our results therefore question whether human health could
be affected by Roundup (10).
The concentration of Roundup that provokes cell cycle dysfunction appears to largely exceed the recommended
usage concentration as an herbicide. The residual concentration of Roundup and/or glyphosate in soil or water is lower, glyphosate being in the nanomolar range present
almost constantly. Cellular dysfunction in the cell cycle occurred at the millimolar range concentration of glyphosate
for minute range exposure. Nevertheless, the discrepancy between the residual doses of glyphosate or Roundup and the
doses inducing dysfunction should not lead one to underestimate the risk. First, the large difference in glyphosate
concentrations could be partially compensated by the great difference in the time exposure. Second, the initial target of
the pesticide in the embryos is not yet identified and might be affected by much lower concentrations of Roundup.
Third, in our experiments, the pesticide affects 100% of the cells whereas cancers and tumors develop from few cells
under the same environmental exposure (21), suggesting potential ef-ficiency of much lower concentrations of the
pesticide.
Roundup contains surfactants, which promote wetting of plant surface and rapid herbicide penetration. In pre-vious
reports, the toxicity of Roundup was ascribed to the surfactant component (12). We show that glyphosate and
formulation products act in synergy on the cell cycle indicating an effect of glyphosate by itself. Our experi-ments
indicate that glyphosate requires the presence of the formulation products to be effective on the embryo. It is likely that
the formulation products favor the pene-tration of glyphosate in the embryos that were already reported to be
impermeable to some compounds (9).
The synthesis of cyclin B is required during the first cell cycle in sea urchin (16, 19, 20). Roundup inhibits the rise
in protein synthesis associated with early develop-ment, without affecting significantly the accumulation of cyclin B as
judged from immunoblots performed with anti-cyclin B antibody. The discrepancy between the inhibition of global
protein synthesis together with the maintenance of specific synthesis of cyclin has already been described using sea
urchin (20). Global protein synthesis is very low in gametes and strongly increases after fertilization (9) due to the derepression of the translation machinery at the level of initiation control (22, 23) and to unmasking of maternal mRNAs
(24). Since the synthesis of cyclin B is not affected significantly, Roundup may hinder, through global protein
synthesis inhibition, the synthesis of a yet unidentified protein that would be necessary to turn on the mitosis switch
and activate the CDK1/cyclin B complex (7). Existence of such protein has been postulated since threshold doses of
emetine can delay cell cycle by inhibiting global protein synthesis without affecting accumulation of cyclin (20). This
hypothetical protein would be involved in the pathway leading to the switch activation CDK1/cyclin B. Roundup could
affect the activation switch by hindering the putative protein action or more directly by impeding with the mechanism
of the switch itself such as dephos-phorylation/phosphorylation activities (7).
Acknowledgment. We thank GerardPeaucellier(Banuyls, France) for the generous gift of anti-cyclin B antibody. We
are very grateful to CecileMaguer, for her expert technical assistance in all the developmental experiments. We address
many thanks to Julia Morales and Patrick Cormier for helpful comments. This work was supported by ConseilRegional de
Bretagne, FondsEuropeens de DeveloppementRegional (FEDER), As-sociation de la Recherchecontre le Cancer (ARC)
and LigueNationalecontre le Cancer.
85
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