Estrogenic Project BI 242 Environmental Science

1. Estrogenic ProjectBI 242 Environmental Science Rachel Coughlin Sarah Laudenslager Meghan Maginnis Michael Southerton

3. The Endocrine System • Controls biological processes • Consists of Glands (Pituitary, Adrenal, Thyroid, Pineal) • Hormones are produced by the Glands • When the Hormones are released they bind with receptors (plasma membrane or intracellular) located in certain organs and tissues. • This binding equals biological activity

4. Effects of Endocrine Disrupters • Endocrine Disrupters are chemicals that interfere with the natural Endocrine System • They “trick” the body into believing they are natural hormones • Mimic • Trigger • Block

5. http://www.mindfully.org/Water/Wastewater-Contaminants-US-StreamsMar02.htmhttp://www.mindfully.org/Water/Wastewater-Contaminants-US-StreamsMar02.htm

6. What is the Government Doing? • In 2003, 2004, 2005, the Bush administration tried to cut all EPA funding for independent scientists who do Endocrine-Disrupting research. • Total Budget for those three years was a combined 15 million dollars. • By comparison, Japan recently spent 135 million dollars on a research program. • http://bb.desales.edu/@@6212558F3460BDB0A498840036A3CF7C/courses/1/BI242-

7. What We Did • Testing of Lehigh Valley Wastewater Treatment Plants • We looked for the concentration of the EDC Estradiol in the influent (pre-treatment water) and the effluent (post-treatment water) over the course of three years. • Upper Saucon • Easton • Philipsburg • Nazareth • Bath • Allentown

8. But First… A little more about EDC’s… EDC Disaster???

9. Insecticides • Pesticides are intentionally released • Operate by killing the insect or otherwise preventing it from engaging in behaviors deemed destructive • Little is known about the extent of environmental transport and fate of these compounds. • Many believed to be Endocrine Disrupting Compounds • Interfere with endocrine systems of humans and wildlife • DES and methoxychlor – delay ovulation in adult rats • Dicofol- retards reproductive development in alligators • Alters concentrations of sex steroid hormones • PCB- alter sexual determination in turtles • Vinclozolin- delay puberty in rats (Crews 2000) (Lyons 1999)

10. Chlordane IUPAC Name: 1,2,4,5,6,7,8,8-octachloro-2,3,3a,4,7,7a-hexahydro-4,7-methanoindene • C10H6Cl8 • First used in the 1940s as an insecticide • Usage was restricted to termite killing in 1978 • Banned in 1988 • Garages and soil still have remnants of previous chlordane usage • Not flammable, but may decompose in flammable solvents • Chlordane decomposes to produce toxic fumes that include chlorine, phosgene (mustard gas), and hydrogen chloride. (Stuart 2001)

11. Transported • Major sources of transport are through the air and water • Enters atmosphere by: soil erosion by wind, dust and sprays • Enters aquatic systems by: surface runoff and rainfall • Absorbed into the bottom sediments • Can last from 3-15 years • Highest samples are found closest to where it was sprayed • Convulsions, seizures, coma, respiratory depression • It is considered a toxic systemic poison that is absorbed well by inhalation, and through the skin • On occasion, has been found in breast milk, as has oxychlordane the stable metabolite. (Eisler 1990) Effects on Humans

12. Studies Exhibiting Endocrine Effects • To determine the combined affect of environmental chemicals on hormonal activity • estrogen receptors from either an Alligator (aER) or Human (hER) were incubated with one of three chemicals: Chlordane, dieldrin, and toxaphene. • Chlordane, dieldrin, and toxaphene individually demonstrated no significant displacement of 17ß-estradiol from aER and hER at the concentration tested. • A combination of these chemicals; however, inhibited the binding of 17ß-estradiol by 20 to 40% (Arnold 1997). • Toxicologist John A. McLachlan did a similar study involving a combination of these three compounds. • Inserted human estrogen receptors into yeast cells and a chemical response system that turned yellow when activated • Chlordane and the compounds, taken individually, are weak when mimicking estrogen • When chlordane was combined with another compound they acted synergistically and created an effect almost as toxic as dieldrin (insecticide endocrine disrupting compound). (Raloff 1996).

13. Effect of endocrine-disrupting chemicals on sex determination in the red eared slider turtle • Analysis of yolk from alligator eggs identified different compounds in different amounts • Same concentrations were applied to red-eared slider turtle eggs • Significant number reversed sexes • Influenced steroid hormone levels after hatching (low compared to the amount of natural estrogen) • Data suggests that even at very low concentrations, endocrine disruptors found in the environment carry risk (Crews 2000).

14. Air Concentrations (ng/m3) Fresh Water Concentrations (ng/L) (Eisler 1990)

15. Seawater Concentrations (ng/L) Soil Concentrations (ng/kg) Sediment Concentrations (ng/kg) (Eisler 1990)

16. Estradiol vs. Chlordane (Gordon 2004) Chlordane is 10,000 times weaker than estradiol

17. Works Cited • Crews, David. (2000, September). Endocrine Disruptors: Present Issues, Future Directions. The Quarterly Review of Biology. 75, 243-260. Retrieved March 19, 2006 from EBSCO.Stuart, Bennent M. (2001). Chlordane. Retrieved March 20, 2006 : http://www.the-piedpiper.co.uk/th13(k).htm • Eisler, Ronald. (1990, July). Chlordane Hazards to Fish, Wildlife and Invertebrates: A Synoptic Review. Contaminant Hazard Reviews. 85, 1-62. Retrieved March 20, 2006: http://www.pwrc.usgs.gov/infobase/eisler/CHR_21_Chlordane.pdf • Arnold, Steven F. (1997, March). in vitro Synergistic Interaction of Alligator and Human Estrogen Receptors with Combinations of Environmental Chemicals. Environmental Health Perspectives. 103, 1. Retrieved March 20, 2006: http://www.ehponline.org/docs/1997/Suppl-3/arnold.html • Lyons, Gwynne. (1999, December). Endocrine Disrupting Pesticides. Pesticides News. 46, 16-19. Retrieved March 19, 2006. http://www.pan-uk.org/pestnews/actives/endocrin.htm • Raloff, J. (1996, June). `Estrogen' pairings can increase potency. Science News. 149, 356. Retrieved March 20, 2006 from EBSCO. • Gordon, John D. Detection of Estrogen Receptor Endocrine Potency of Commonly Used Organochlorine Pesticides Using the LUMI-CELL Bioassay. Organohalogen Compounds. 66, 196-174. Retrieved March 20, 2006: http://www.dioxins.com/pdfdocs/LUMI-CELL%20ER%20Pesticides%20Dioxin%202004.pdf

18. Plasticizers Danger may lurk in the most unexpected places.

19. Plasticizers • Plasticizers:Small, often volatile molecules that are added to hard, stiff plastics to make them softer and more flexible. • From http://www.epa.gov/envirohealth/children/background/glossary.htm

20. Types of Plasticizers • Pthalate-based Plasticizers • Bis(2-ethylhexyl)phalate (DEHP) • Diisononyl phthalate (DINP) • Butyl benzyl phthalate (BBzP) • Bis(n-butyl)phthalate (DnBP, DBP) • Diisodecyl phthalate (DIDP), • From http://www.answers.com/topic/plasticizer

21. More Types of Plasticizers • Adipate-based plasticizers • Used for low-temperature or resistance to ultraviolet light • Bis(2-ethylhexyl)adipate (DOA) • Dimethyl adipate (DMAD) • Monomethyl adipate (MMAD) • Dioctyl adipate (DOA) • From http://www.answers.com/topic/plasticizer

22. Still More Types of Plasticizers • Trimellitates • used in automobile interiors and other applications where resistance to high temperature is required • Trimethyl trimellitate (TMTM) • Tri-(2-ethylhexyl) trimellitate (TEHTM-MG) • Tri-(n-octyl,n-decyl) trimellitate (ATM) • From http://www.answers.com/topic/plasticizer

23. Other Plasticizers • Maleates • Benzoates • Epoxidized Vegetable Oils • Sulfonamides • Phosphates • Glycols / Polyethers • From http://www.answers.com/topic/plasticizer

24. Phthalates • Most Common type of Plasticizer • “Phthalates migrate to the surface of plastics, and can then evaporate or leach into the surrounding environment. This limits the usefulness of phthalate plasticizers, as they eventually migrate out of plastics entirely, and as a result the plastics become brittle 2. The release of phthalates into the environment represents an environmental hazard as well. Because of the widespread use of phthalates, they have become one of the most abundant industrial pollutants in the environment.” • From http://www.carbohydrateeconomy.org/library/admin/uploadedfiles/Biochemical_Plasticizers.html • Photo from http://www.answers.com/topic/phthalates

25. Food Packaging Children’s Toys Medical Devices Cling Wrap Vinyl Tiles Traffic Cones Food Conveyer Belts Artificial Leather Plastic Foams From http://www.answers.com/topic/plasticizer Shoes Garden Hoses Building Materials Cellulose Plastics Food Wraps Adhesives Perfumes Cosmetics Where are Phthalates Found?

26. Health Concerns • Health Care Without Harm • “Aggregate Exposures to Phthalates in Humans” – July 2002 • Analyzed and exposed many concerns regarding human contact with phthalates • http://www.noharm.org/details.cfm?type=document&id=796

27. Health Care Without Harm FindingsWho is exposed? • Young, developing organisms are more vulnerable to exposure to phthalates than adults • In particular, the developing male reproductive tract appears to be the most sensitive to exposure to several phthalates. • Effects on the liver, kidneys, lungs, and blood clotting are also of concern. • Women of reproductive age experience some of the highest exposure levels to phthalates - a special concern because phthalates can interfere with normal reproductive tract development of developing boys during pregnancy. • Children also have elevated levels of some phthalates.

28. Health Care Without Harm FindingsHow are people exposed? • Direct Exposure • Direct use of cosmetics or exposure to medical devices containing Phthalates • Indirect Exposure • Puts general population at risk • Exposure through discarded products, causing “Worldwide Ecosystem Contamination.”

29. Phthalate Exposure and EarlyThelarche • In 2000, Colon, I, D Caro, CJ Bourdony, and O. Rosario. Released their findings that that since 1969 4,674 cases of Thelarche (the premature growth of breasts in girls) in Puerto Rico. This included the development of breast in a 23 month old child. • They documented the Thelarche as being a result of Phthalates present in the water supply. • http://bb.desales.edu/webapps/portal/frameset.jsp?tab=courses&url=/bin/common/course.pl?course_id=_3897_1

30. Dangerous Levels • The June 1998 edition of Consumer Reports sought to find the migration of phthalates from plastic wrappers to cheese slices. • The study provides an indication of the danger of Phthalates. • They identified three levels of contamination: • Very High (50 to 160 parts per million) • Moderate (1 to 4 parts per million) • Little to No Contamination • From www.mindfully.org/Pesticide/Hormone-Mimics-In-Food.htm

31. Avoiding Phthalates • http://www.charityadvantage.com/mhop/whatyoucandotoreduce.asp

32. Personal Care Products: phthalates DEHP di(2-ethylhexyl) Homer doesn’t allow cosmetics under his roof…should you?

33. Chemicals DEHP is a plastcizer in chemical form: di(2-ethylhexyl Phthalic Acid, Di(2-ethylhexyl) EsterMolecular Formula: C24H38O4Abbreviations: DEHP, MEHP [mono(2-ethylhexyl) phthalate], PVC (polyvinyl chloride), PP (peroxisome proliferation).Synonyms and Trade Names: 1,2-Benzenedicarboxylic acid, bis(2-ethylhexyl) ester; Bis(2-ethylhexyl) phthalate; Compound 889; Di(2-ethylhexyl) orthophthalate; Di(2-ethylhexyl) phthalate; Ergoplast FDO; 2-Ethylhexyl phthalate; Vestinol AH; Vinicizer 80; Witcizer 312

34. Molecular formulae:

35. Physical and Chemical Properties of DEHP • DEHP is a colorless liquid with almost no odor • Solubility: insoluble in water • Water 41 μg/L at 25 ECa • Organic solvent(s) Miscible in mineral oil and hexane • Soluble in most organic solvents

36. Potency Compared to Estradiol Estradiol DEHP

37. Potency (cont.) • Di(2-ethylhexyl) phthalate (DEHP) showed no estrogenic activity within in vitro • phthalate plasticiser DEHP (di-2-ethylhexyl phthalate) measured estrogenic potency is very weak

38. Sources • DEHP is widely used in medical devices, flooring, auto parts, and many other products. • DEHP is a phthalate ester widely used as a plasticizer to make polyvinyl chloride (PVC or vinyl) medical products soft and flexible and to increase moisturizing properties of some personal care products • It is used to detect leaks in protective face gear, and as a test material for filtration systems. It has also been used as a replacement for polychlorinated biphenyls (PCBs) in dielectric fluids for electric capacitors and in vacuum pumps. • According to EPA’s Toxics Release Inventory for 2000, reporting industries released nearly 250,000 pounds of DEHP into the environment.

39. Sources (cont.) • Point sources-from emissions or spills from sites that use DEHP in their manufacturing processes.Diffuse sources, and point sources included in aggregated emissions data-its wide use, volatility, & persistence mean that DEHP is widely distributed in the environment.Natural sources-DEHP has been suggested as a possible natural product in some animals and plants.Mobile sourcesNone Consumer products which may contain Di-(2-Ethylhexyl) phthalate (DEHP)-DEHP is in polyvinyl chloride (PVC) plastic products like toys, vinyl upholstery, shower curtains, adhesives, and coatings. It is used in some food packaging, and medical product containers (including those for blood) and equipment. It is also used in some inks, pesticides, cosmetics, and vacuum pump oil

40. Effects; why are they hard to determine? • Humans are exposed to substantial levels of DEHP through medical devices. Hemophiliacs, kidney dialysis patients, and high risk newborns are particularly heavily exposed. • The few studies of exposed human populations means that conclusions about DEHP risks must be based on laboratory animal studies. • Studies of laboratory animals, supported by very limited human data, suggest that a wide range of toxic effects occur in exposed mammals. Insufficient evidence exists to conclude that the toxic mechanisms found in laboratory animals do not occur in humans. • Uncertainty about the potential health hazards of DEHP remains. Quantitative estimates of risk to humans at various stages of life or health, or of safe levels of exposure, cannot be validated at this time. • Materials exist which do not contain DEHP or similar plasticizers, and are currently being used in medical devices. These materials are potentially safer alternatives to DEHP-containing medical devices.

41. Effects on Humans • Health effectsExposure: from use of medical products packaged in plastic such as blood products (particularly when used extensively, such as for kidney dialysis), eating some foods packaged in certain types of plastics or coated papers (ex. fatty foods, milk products, fish and seafood), soils, drinking contaminated water or breathing air containing DEHP where it is used or spilled. Di-(2-Ethylhexyl) phthalate (DEHP) enters the body:DEHP enters the body by breathing the vapors or ingesting it directly or through contaminated products. Absorption through skin contact is also possible but slow.Relative health hazardOn a health hazard spectrum of 0 - 3 Di-(2-Ethylhexyl) phthalate (DEHP) is a 1.3. A 3 represents a very high hazard, 2 represents a medium hazard and 1 is harmful to health. Factors for this ranking are; the extent of the material's toxic or poisonous nature and/or its lack of toxicity, and the evaluation of its tendency to cause, or not cause cancer and/or birth defects. It does not take into account exposure to the substance. • There is no evidence that DEHP causes serious health effects in humans. Most of what we know about the health effects of DEHP comes from high exposures to rats and mice which may not be representative of the effects on humans. Effects observed in animals were from very high and prolonged doses. Exposure to DEHP in air did not result in any observed effects. Exposure in food and water resulted in effects on sperm production, the ability to reproduce and birth defects. Kidney damage similar to the damage seen in the kidneys of long-term dialysis patients has also observed.DEHP is one of a range of phthalates which have been suggested as being able to be effect human and animal endocrine systems (endocrine disruptors)

42. Effects of Wildlife, Environment, & Water and Soil • Environmental effectsDEHP in the atmosphere is present either as a gas or attached to solid particles. It breaks down quickly ( 1-2 days) due to the action of other chemicals in the atmosphere. The solid particles are removed from the atmosphere in 2-3 weeks by precipitation, wash out by rain, and reaction with other chemicals. DEHP is slightly prevalent in the environment. Small organisms in surface water or soil break it down into harmless compounds. It doesn't break down easily in deep soil, or in lake or river bottoms. It is in plants, fish, and other animals, but animals high on the food chain are able to breakdown DEHP, so tissue levels are usually low.Phthalate is slightly present in water but will break down in a few months.Environmental TransportDEHP from plastic materials, coatings, and flooring can increase indoor air levels. It dissolves faster in water if gas, oil, or paint removers are present. DEHP in the particle-phase is subject to wet and dry deposition. It will be transported in food chain & broken down and does not usually bioaccumulate. Concentration of DEHP in fish is expected to be much higher then the concentration in water in which the fish live.About 42.8% of DEHP will eventually end up in terrestrial soil; about 40% will end up in aquatic sediments; and about 17% will end up in air.Relative hazard to the environmentOn an environmental spectrum of 0 - 3 Di-(2-Ethylhexyl) phthalate (DEHP) is 1.2. A 3 represents very high hazard to the environment and 0 a negligible hazard. Factors include the extent of the material's toxic or poisonous nature and/or its lack of toxicity, and the measure of its ability to remain active in the environment and whether it accumulates in living organisms. It does not take into exposure to the substance.

43. Phthalates as Endocrine Disruptor Compounds: • a very significant relationship between a mother's exposure during pregnancy to phthalates changes in the ways that baby boy's genitals develop. • Baby boys with greater phthalate exposure had smaller AGI (anogenital index). measurements than normal. • Boys exposed to multiple phthalates simultaneously were also more likely to have smaller AGI scores. • Boys with lower AGIs had smaller penis volumes and were more likely to experience incomplete testicular descent. • The FDA warns anyone who is exposed to PVC (polyvinyl chloride) in a medical setting. DEHP is released from medical devices into solutions that come in contact with this plasticizer. Alternative methods are suggested when high exposure must take place; Especially for male neonates, pregnant women carrying male fetuses and peripubertal males (all having the greatest risk of negative effects).

44. EDC’s (cont.) • A valid human study linked prenatal phthalate exposure to adverse effects on the male reproductive system. • The associations between prenatal phthalate exposure and decreased AGI were very large and highly statistically significant. • In rodents, fetal exposure to phthalates causes a “phthalate syndrome.” It includes decreases in AGD, increased frequency of incomplete testicular descent and a birth defect of the penis called 'hypospadias,' increased risk of testicular cancer in adulthood, impaired sperm quality and more. • Scientists studying people found a pattern of male abnormalities called testicular dysgensis syndrome.' It includes hypospadias, cryptorchidism, impaired sperm quality and testicular cancer. One potential cause of TDS has been exposure to endocrine disrupting chemicals like phthalates. • The changes in AGI were observed at phthalate levels below those found in one-quarter of women in the United States, based on CDC’s nationwide sample. • Women are also denying products with any chemical link to defects (ex. Phthalates) • One of the key points is DEHP's impact on developing Sertoli cells, cells in the male reproductive tract that are central to sperm formation. Damaged Sertoli cells during development lead to sperm maladies in adulthood, including low sperm count. DEHP does not cause Sertoli damage directly; damage is caused by a metabolite of DEHP, monoethylhexyl phthalate (MEHP).

45. Sources Used: www.epa.gov www.fda.gov http://www.atsdr.cdc.gov/toxprofiles/tp9-c4.pdf http://www.mindfully.org http://www.noharm.org http://www.envirohealthaction.org http://www.npi.gov http://lb.chemie.unihamburg.de/static/data2/92_1upvg4mr.html

46. Detergents • Essential to personal and public health • Contribute to good personal hygiene • Reduce germs that cause infectious diseases • Four categories of detergents • Personal cleansing • Laundry • Dishwashing • Household cleaning

47. Chemicals found in Detergents • There are many different chemicals that are found in cleaning detergents • Acetone • Benzaldehyde • Benzyl Acetate • Linalool • Nonylphenol • Ethylene Oxide

48. Nonylphenol • Part of the alkylphenol family • Used as a surfactant in cleaning and cosmetic products and as a spermicide in contraceptives • C15H240 • Properties • Thick, light yellow, color liquid with slight characteristic phenolic odor • Poorly soluble in water, soluble in alcohols • Effects • Shown to have estrogenic properties • Poisoning decreased weight gain, caused changes in the liver, hemorrhages • Stimulates breast tumor cells in vitro • Increase uterine weight • Allergenic effect-skin sensitization reaction • Safe level in drinking water– 0.001mg/l

49. Nonylphenol (cont.) • Gets into water through everyday actions • Washing hands • Doing the laundry • Taking showers and baths • Have been shown to have effects on many aquatic organisms (bound estrogen receptors) • Non-occupational exposure does not pose any risk for humans • To get the same effect with 17B estrodial you need 11,000 times higher concentration of nonylphenol • Other effects • Human males over the past 2 generations have 50% lower sperm counts • Rising rates of testicular cancer and birth defects (undescended testicles) • Change of genitals in animals

50. Estradiol • Sex hormone labeled as the female hormone but also found in men because it represents the major estrogen in humans • Critical for sexual functioning • Supports bone growth • Derived from cholesterol • Androstendione – key intermediary; is converted to testosterone which undergoes aromatization and then turns into estradiol • Chemical Formula – C18H24O2