bi 242 environmental science
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BI 242 Environmental Science. Environmental Endocrine Disruptors (EDC’s) Dr. Joseph Colosi DeSales University. The Endocrine system. General Features of the endocrine system:. Transport. Gland. Hormone. Target Cell. rich blood supply. hormone receptors are very specific. - PowerPoint PPT PresentationTRANSCRIPT
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BI 242 Environmental Science
Environmental Endocrine Disruptors (EDC’s)
Dr. Joseph ColosiDeSales University
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The Endocrine system
GlandTransport
Target Cell
Hormone
General Features of the endocrine system:
ductless
rich blood supply
secreted into the blood
can reach virtually every cell in the body
hormone receptors are very specific
C. Corbitt
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Endocrine Glands
don’t forget the heart, placenta, fat
All of these glands produce hormones and are also targets for hormones
C. Coebitt
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Insulin, a protein hormone
Steroid hormones
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Link to diagram showinglocations of the endocrine glandsThyroid-stimulating hormone (TSH)protein (201)Anterior lobe of pituitaryFollicle-stimulating hormone (FSH)protein (204)Luteinizing hormone (LH)protein (204)Prolactin (PRL)protein (198)Growth hormone (GH)protein (191)Adrenocorticotropic hormone (ACTH)peptide (39)Antidiuretic hormone (ADH)(vasopressin)peptide (9)Posterior lobe of pituitaryOxytocinpeptide (9)Thyrotropin-releasing hormone (TRH)peptide (3)HypothalamusGonadotropin-releasing hormone (GnRH)peptide (10)Growth hormone-releasing hormone (GHRH)peptides (40)Corticotropin-releasing hormone (CRH)peptide (41)Somatostatinpeptides (14, 28)DopamineTyrosine derivativeMelatoninTryptophan derivativePineal glandThyroxine (T4)Tyrosine derivativeThyroid GlandCalcitoninpeptide (32)Parathyroid hormone (PTH)protein (84)Parathyroid glandsGlucocorticoids (e.g., cortisol)steroidsAdrenal cortexMineralocorticoids (e.g., aldosterone)steroidsAndrogens (e.g., testosterone)steroidsAdrenaline (epinephrine)Tyrosine derivativeAdrenal medullaNoradrenaline (norepinephrine)Tyrosine derivativeEstrogens (e.g., estradiol)steroidOvarian follicleProgesteronesteroidCorpus luteum and placentaHuman chorionic gonadotropin (HCG)protein (237)Trophoblast and placentaAndrogens (e.g., testosterone)steroidTestesInsulinprotein (51)Pancreas (Islets of Langerhans)Glucagonpeptide (29)Somatostatinpeptides (14, 28)Amylinpeptide (37)Erythropoietin (EPO)protein (166)KidneyCalcitriolsteroid derivativeCalciferol (vitamin D3)steroid derivativeSkinAtrial-natriuretic peptide (ANP)peptides (28,32)HeartGastrinpeptides (14)Stomach and intestineSecretinpeptide (27)Cholecystokinin (CCK)peptides (8)Somatostatinpeptides (14,28)Neuropeptide Ypeptide (36)Ghrelinpeptide (28)PYY3-36peptide (34)Insulin-like growth factor-1 (IGF-1)protein (70)LiverAngiotensinogenproteinThrombopoietinprotein (332)LeptinproteinFat cellsNote (1): Numbers within parentheses indicate the number of amino acids in the protein or peptide(s).
There are many hormones
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Types of hormone receptors
R
Enzyme effector
R G-protein
Enzyme effector
R Ion channel
HORMONE
NUCLEUS
R
R
DNA binding sites
Nuclear membrane
CELL MEMBRANE
Protein hormones
Intracellular Receptors
Plasma Membrane Receptors
Steroid hormones
C. Corbitt
Protein product
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HYPOTHALAMUS
PITUITARY
TESTIS
Testosterone
The H-P-G Axis
-
-
- = negative feedback
GnRH-
- LH
C. Corbitt
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It’s actually much more complicated
C. corbitt
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Do exogenous hormones affect people?
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Diethylstilbestrol (DES)
• 1938 Charles Dodds synthesized DES• 1941 Harvard: DES enhances pregnancy and prevents miscarriage• 1947 FDA Approved DES for pregnancy• 1947-1971 DES prescribed for 5 million US pregnant women, dose =
700 birth control pills• 1953 U of Chicago study: no DES miscarriage benefit
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Diethylstilbestrol (DES)
• 1954 DES put in chicken and cattle feed• 1959 DES banned for chickens and lambs• 1964 Charles Dodds Knighted• 1971 Surgeon General warned against DES for pregnant women• 1979 USDA banned DES for cattle feed• 1970’s DES prescribed to many thousands of pregnant women
throughout the world
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DES Outcomes (CDC DES page http://www.cdc.gov/des/consumers/)
DES Daughters CDC DES daughter page
http://www.cdc.gov/des/consumers/about/effects_daughters.html
• One third have reproductive tract abnormality • 100X greater risk for clear cell adenocarcinoma, from 0 to 1/1000• 2.5X greater chance of miscarriage• 2 to 3X greater chance of ectopic pregnancy• 33% increase in infertility• 25% increase in premature delivery
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DES Outcomes (http://www.cdc.gov/des/consumers/)
DES Sons• 4X increase in non-cancerous epidymal cysts• 3 to 4X increase in genital deformity?
DES Grandchildren?
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So avoid high doses of steroid drugs and you’ll be safe.
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First nationwide survey to detect 95 pharmaceuticals, hormones, and other organics in 139 urban and agricultural streams in 30 states. Two chemicals found in 80% of the samples, and 82 chemicals occurred in at least one sample.
USGS Water-quality survey, 1999-2000
Flame retardant
Insect repellant
Fungicide
antibacterial
surfactant
surfactant
surfactantplasticizer
Survey study 2002 http://pubs.acs.org/hotartcl/est/es011055j_rev.html
New study on pesticides 2006 http://pubs.usgs.gov/fs/2006/3028/
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US
GS
Wa
ter-
qu
alit
y d
ata,
19
99-
200
0
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PPB
PPT
Detection limits for USGS study
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Compounds known to bind steroid receptors in humans and animals
Rooney, AA, and LJ Gillette, Jr. Contaminant interactions with steroid receptors: evidence for receptor binding. In Guillette, LJ, Jr., and DA Crain. 2000 Environmental Endocrine Disrupters: An
Evolutionary Perspective. Taylor and Francis.
• Diethylstilbestrol (DES) Food contaminant• Butylated hydroxyanisole Food additive• Vinclozolin fungicide• Alachlor Herbicide• DDT Insecticide• P-Nonylphenol Industrial chemical• Genistein Phytoestrogen
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Compounds known to bind steroid receptorsRooney, AA, and LJ Gillette, Jr. Contaminant interactions with steroid receptors: evidence for receptor binding. In Guillette,
LJ, Jr., and DA Crain. 2000 Environmental Endocrine Disrupters: An Evolutionary Perspective. Taylor and Francis.
Pesticides Industrial chemicals PhytoestrogensVinclozolin
Alachlor
Atrazine
Cyanazine
Desiopropyl
Simazine
Prometryne
o,p’-DDD
p,p’-DDD
p,p’-DDE
o,p’-DDE
o,p’-DDT
p,p’-DDT
DDOH
2,4-Dichlorophenol
Chlordecone (Kepone)
Dicofol
Endosolfan sulfate
-Endosolfan
Methoxychlor
cis-Nonachlor
trans-Nonachlor
Toxaphene
Tamoxifen (drug)
Aroclor 1242 (PCB
mixture)
p-sec Amyl phenol
p-sec-Amyl phenol
p-Isoamyl phenol
p-tert Amyl phenol
o-sec-amyl phenol
Bensophenone
Bisphenol-A
bis(2-Ethylyexyl) phthalate
bis(2-Ethylyexyl) adipate
6-Bromonaphthol-2
4-sec-Butylphenol
n-butyl-benzene
Di-n-Butylphthalate
Butylbenzyl phthalate
4,4’-Dihydroxy
2’chlorobiphenyl
4,4’-Dihydroxybiphenyl
4-Hydroxy, 2’,4’,6’-
trichloro biphenyl
4-Hydroxy, 2’,3’,4’,5’-
tetrachloro biphenyl
1-Naphthol
2-Naphthol
4-nitrotoluene
p-Nonylphenol
Nonylphenol
4-Nonylphenoxy carboxylic acid
4-tert-Octylphenol
-Phenylcresol
Tetrahydronaphthol-2
3,4,3’,4’-tetrachloro biphenyl
Anglolensin
Apigenin
Biochanin-A
Chrysin
Coumestrol
Daidzein
o-Desmethylangolensin
4,4’Dihydroxychalcone
4,7’-Dihydroxyflavanone
Equol
Genistein
Isoliquiritigenin
Kaempferide
Luteolin
Miroestrol
Naringenin
Phloretin
-Sitosterol
Zearalenone (Mycoestrogen)
Zearalenone (Mycoestrogen)
Zearalanol (Mycoestrogen)
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Every year, 5000 new chemical compounds are introduced.
“Better living through chemistry.”
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How do these chemicals get into the environment?
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What it takes to be beautiful today.
Where do these chemicals go when we shower and pee?Why pee?
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Bethlehem Sewage Treatment Plant
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Plastics
R. Bolen
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Cattle feedlot Sheep feedlot
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Pesticides
R. Bolen
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Industrial chemicals
R. Bolen
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Do these chemicals in the environment affect wildlife?
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There are hundreds of research articles relating EDC’s to effects on wildlife:• feminization of male alligators, tadpoles
• masculinization of female gastropods; female fish
• impaired immune and decreased reproductive function in Baltic seals
• eggshell thinning in water birds, eagles• all have resulted in population decline.
R. Bolen
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Reproductive and developmental abnormalities attributed to endocrine disruption
from McLachlan (2001)
Species Observation Contaminant
alligators abnormal gonads, decreased phallus size, altered sex hormone levels
DDT, DDE, dicofol
snails masculinization,, imposex, formation of additional female organs, malformed oviducts, increased oocyte production
tributylin, bisphenol A, octylphenol
seals impaired reproductive function coumestrol
water birds egg shell thinning, mortality, developmental abnormalities, growth retardation
DDE, PCBs, AhR agonists
mosquito fish abnormal expression of secondary sex characteristics, masculinization
androstenedione
R. Bolen
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McLachlan, J.A. 2001 Environmental Signaling: What Embryos and Evolution Teach Us About Endocrine Disrupting Chemicals, Endocrine Reviews 22(3): 319-341.
How Do Environmental EDC’s Work?
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Do these chemicals in the environment affect humans?
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Trends in human reproductive health• testicular dysgenesis syndrome
– decreased semen quality (reduced numbers, motility, altered morphology of sperm)
– increased frequency of male reproductive tract abnormalities• hypospadias (urinary tract opening located not at the tip of the penis)• cryptorchidism (testis fails to descend by age 12 months)
• altered sex ratio (fewer male babies)• premature breast development (thelarche)• premature onset of menstruation (menarche)• increase in rates of breast cancer, testicular cancer
Are these trends In humanscaused by EDC’S?
R. Bolen from Sharpe and Irvine (2004)
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What evidence links human reproductive problems to EDCs?
• Girls exposed to higher levels of PBCs and DDE in utero entered puberty an average of 11 months earlier than controls
• Higher levels of organochlorine chemicals found in mothers of men with testicular cancer
• Exposure of boys to endosulfan associated with delayed puberty• Premature thelache in Puerto Rico associated with high blood
phthalate levels.• Prenatal blood phthlate associated with feminization of male babies.
R. Bolen
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Premature Thelarche
23-month-old Puerto Rican girl with premature thelarche
Thelarche: breast development, timing depends on estrogen/androgen ratio
Premature thelarche “epidemic” in Puerto Rico
Premature Thelarche and Early Sexual Development Registry tracked 4,674 cases in P.R. since 1969
Several causes hypothesized: diet, EDCs
Colon, I, D Caro, CJ Bourdony, and O.Rosario. 2000. Identification of phthalate esters in the serum of young Puerto Rican girls with premature breast development. Environ. Health Perspect. 108:895-900
Plasma phthalate ester levels correlated with premature thelarche cases
Phthalate esters used as plasticizers; have estrogenic or anti-androgenic activity in vitro
C. Corbitt
But, McKee, RH. 2004. Phthalate exposure and early thelarche. Environ. Health Perspect. 112:A541-A543. Questions the results because the blood levels are too high and phthalates do not act like estrogens in female rodents
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penis
Anogenital distanceAGD
AGI = AGD/weight
. August 2005
anus
Environmental Health Perspectives 113: 1056-1061
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What is EPA doing about it?
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EPA Screening protocols being testedTier 1 Screening will include a battery of screening assays that would identify substances that have the potential to interact with the estrogen, androgen, or thyroid hormone systems.
Tier 2 Testing AssaysWill determine whether a substance may cause endocrine-mediated effects through or involving estrogen, androgen, or thyroid hormone systems,
Will determine the consequences to the organism of the activities observed in Tier 1
Will establish the relationship between doses of an endocrine-active substance administered in the test and the effects observed.
Amphibian Metamorphosis
(Tadpoles)
Androgen Receptor (AR) Binding
Aromatase
Estrogen Receptor (ER) Binding
Fish Screen (Whole fish)
Hershberger (Whole rats)
Pubertal Female (Whole rats
Pubertal Male (Whole rats)
Amphibian 2 (Whole frogs)
Avian 2-Generation (Whole birds)
Fish Lifecycle
Invertebrate Lifecycle (Whole mysid
shrimp)
Mammalian 2-Generation (Whole rats)
Tier (to be determined)
In Utero through Lactation (Whole rats)
http://www.epa.gov/scipoly/oscpendo/edspoverview/chronology.htm 1996-2005
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EPA will test for pesticide concentration in the the 20 most consumed foods for pesticides
http://www.epa.gov/fedrgstr/EPA-TOX/2005/September/Day-27/t19260.pdf
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Current US regulations on environmental EDC’s
•Banned: aldrin, chlordane, DDT, dieldrin, endrin, kepone, mirex, PCB’s, toxaphene•Restricted: dicofol, dienochlor, endosulfan, heptachlor, lindane, methoxychlor•Not regulated: Vast majority of EDC’s.
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Yeast Bioassay for Estrogenic CompoundsEstrogen-inducible expression system in yeast
Routledge, EI, JP Sumpter. 1996. Estrogenic activity of surfactants and some of their degradation products assessed using a recombinant yeast screen. Environmental Toxicology and Chemistry 15: 241-248
Sensitive to 10 PPT 17 -estradiol
1 part per trillion is one second in 32 thousand years.
A playing card on a football field is one part per million. Cut the card into 1 million pieces. One of these pieces on a football field is one part per trillion.
ONPG
(Clear)
Yellow
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PPB
PPT
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Witters et al. (2001) Method for Extraction of Estrogenic Compounds from Wastewater
2.5 Liters of wastewater passed through 3 sets of filters (final pore size 0.45μm), then solid phase extraction disk. Estrogenic compounds eluted with acetone, dried with nitrogen, and dissolved in 100μl DMSO.
Revised method -0.2μm sterile filter
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Standard Curve for Quito 10/27/2005
y = -2E+30x3 + 3E+20x2 + 6E+09x - 0.0092
R2 = 0.9999
0
0.5
1
1.5
2
2.5
0.00E+00 2.00E-11 4.00E-11 6.00E-11 8.00E-11 1.00E-10 1.20E-10 1.40E-10 1.60E-10
Estradiol Molar Concentration
OD
405
Estrogenic compounds stimulate the yeast to convert the ONPG substrate into a yellow pigment. More yellow signifies more estrogenic compounds in the water.
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Estradiol Equivalents in Six Lehigh Valley, PA Wastewater Treatment Plants
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
Pberg Easton Usaucon Nazareth Bath Allentown
Wastwater source
Est
rad
iol
Eq
uiv
alen
t,
Par
ts p
er T
rill
ion 7/20/04 Influent
7/20/04 Influent
10/8/04 Influent
10/8/04 Effluent
3/14/05 Influent
3/14/05 Effluent
6/28/05 Influent
6/28/05 Effluent
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•What is the daily pattern of estrogenic chemicals in influent wastewater?•What is the seasonal pattern?•Do these vary with source of wastewater?•How much is the concentration reduced by the treatment? Are some treatments more effective?•Do the levels in the effluent affect aquatic wildlife? Terrestrial wildlife? Humans?
For further study:
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Summary
•The endocrine system is complex and depends on extremely low concentrations of hormones.•Many chemicals are found in US surface waters.•Many of these are EDC’s.•There are disturbing trends in sexual and devolpmental dysfunction in wildlife and humans.•Progress on detection of EDC’s is slow.•Transgenic yeast has potential to expand our knowledge about the occurrence of EDC’s.