Agenda

3M/EPA Science Seminar, April 4

• Introductions/Sign Introductions/Sign‐in ‐ All (5 min)• Introductory Remarks ‐ LZ/JS (5 min)• Comments On Grandjean Et Al. Study Of Immune Respo

Markers in Faroese Children (LRZ, 10 min)

• Is Clearance A Common Explanation Across The Epidem

Associations Associations Reported Reported At “Low” PFOA And PFOS Concen

Concen

(GWO, 120 min)

• Dietary Factors As Potential Effect Modifiers In Epidemio

Studies (SC, 20 min)

• General Discussion

• Adjourn
Comments on Grandjean Grandjean et 

Serum Vaccine Antibody Concentratio

in Children Exposed to Perfluorinated

Compounds

JAMA, Jan 25, 2012, 307(4): 391‐397
Brief Summary

• 656 consecutive singleton births in Faroe I

1999 – 2001

• Vaccinations at 3,5 and 12 months against 

tet, pert, polio and HfluB; DT booster booster age 

booster after 2003; titers measured age 5 

booster, 4 weeks post booster and age 7

• PFOS, PFOA, PFHxS, PFNA and PFDA meas

maternal sera week 32 and children age 5 

booster

• GM PFOS was 27.3 ng/mL in maternal sera 

16.7 ng/mL in children at age 5
Brief Summary

• Actual data (t te s i r v. fluo oc e ca rochemical

concentrations) not presented

• Results of model showing impact of twofo

change in each fluorochemical on titers is 

• Results of a “structural equation model” u

PFOS, PFOA and PFHxS together also show

tabular form.

• Odds ratio for f lli a ng b l e ow prot ti ec ve l 

eve 

antibodies against diphtheria and tetanus 

5 and 7 is presented presented in the text
Brief Summary

• In their model, a doubling of PFOS sera concen

either in maternal or child sera tended to nega

impact titers (14 neg and 2 pos slopes), but the 

included the null in 13 of 16 comparisons

• For PFOS: the odds ratio for falling below the

protective level of diphtheria antibody at age 5 

positive positive and excluded excluded the null based on either

maternal or child sera measurement; the odds 

included the null at age 5 for tetanus titers falli

below the protective protective level, and also included included th

for this effect at age 7 for both tetanus and dip

titers
Comments

• No case of diphtheria reported in Denmark sin

(WHO)

• The authors inexplicably fail to cite Fei et al, 20

found no impact of maternal maternal PFOS concentrati

concentrati

hospitalization for infectious disease up to age 

similar size Danish cohort

• The 3M episodes episodes of care study showed no incr

incr

risk for any infectious disease episode of care i

chemical v film plant employees between 1993 

1998

• SOT poster – lack of impact on infectious disea

• No justification for g pg rou ing fluorochemicals
Comments

• The immunotoxicology literature for PFOS is no

accurately represented; they state that immuneffects have been found in mice at “...serumconcentrations concentrations similar similar to those reported reported in the 

study...” (27 and 16 ng/mL)

• Peden‐Adams is reported an effect on humera

immunity at 92 ng/mL; Qazi could not replicate 

result and reported this effect at over 11,000 n

• No other study in which PFOS sera concentrati

concentrati

measured reported an effect on an immune en

at less than 600 ng/mL
Is Clearance a Common Explana

Across the Epidemiological 

Associations Reported at “Low

PFOA and PFOS Concentration

Geary Olsen*, Sue Chang, John Butenhoff, 

Carol Ley, Larry Zobel

April 4, 2012

*Presenter
Biomonitoring
30.4

34.9

253035404550

Concentration (ng/mL)

Geometric Mean Perfluoroalkyl Biomonitoring Trends in NHAN

and the American Red Cross Studies, 1999 – 2010

Olsen et al. Environ Sci Technol 2012 (submitted)

2.1 2.25 1.93 1.67 1.52 1.96 

20.7

17.1

14.5 13.2

5.2 4.7 3.95 3.92 3.44 4.1

05

101520

25

NHANES 

1999-2000

Am Red 

Cross 2000-

2001

NHANES 

2003-2004

NHANES 

2005-2006

Am Red 

Cross 2006

NHANES 

2007-200

Concentration (ng/mL)
30.4

34.9

3035404550

Concentration (ng/mL)

Geometric Mean Perfluoroalkyl Biomonitoring Trends in NH

and the American Red Cross Studies, 1999 – 2010

Olsen et al. Environ Sci Technol 2012 (submitted)

2.1 2.25 1.93 1.67 1.52 1.96 

20.7

17.1

14.5 13.2

5.2 4.7 3.95 3.92 3.44 

05

10152025

NHANES 

1999-2000

Am Red Cross 

2000-2001

NHANES 

2003-2004

NHANES 

2005-2006

Am Red Cross 

2006

NHAN

2007-20

Concentration (ng/mL) 
30.4

34.9

20.7

253035404550

Geometric Mean Biomonitoring Trends in NHANES

and the American Red Cross Studies, 1999 – 2010

NEW NHANES DATA Supports American Red Cross 2010 Analyse

New

NHANES

Data

2.1 2.25 1.93 1.67 1.52 1.96 1.52 

20.7

17.1

14.5

13.2

8.7 

5.2 4.7 3.95 3.92 3.44 4.13 2.84 

05

101520

NHANES 

1999-2000

Am Red 

Cross 2000-

2001

NHANES 

2003-2004

NHANES 

2005-2006

Am Red 

Cross 2006

NHANES 

2007-2008

NHANES 

2009-2010
20

253035404550

Concentration (ng/mL)

Geometric Mean Perfluoroalkyl Biomonitoring Trends by Loca

American Red Cross Studies, 1999 – 2010

Olsen et al. Environ Sci Technol 2012 (submitted)

05

1015

20

Boston 2000-2001

Boston 2006

Boston 2010

Charlotte 2000-2001

Charlotte 2006

Charlotte 2010

Hagerstown 2000-2001

Hagerstown 2006

Hagerstown 2010

Los Angeles 2000-2001

Los Angeles 2006

Los Angeles 2010

Mpls-St. Paul 2000-2001

Mpls St. Paul 2006

Mpls-St. Paul 2010

Concentration (ng/mL)
CONCLUSIONS (Olsen et al. Environ Sci Technol 2012 , submitted)

• Population halving time for PFOS is approximately 4.3 to 5.0 years.• This is similar to the individual geometric mean serum elimination half-life o• This similarity suggests the dominant PFOS-related exposures to humans in t

were greatly mitigated during the phase-out period.

• The PFOA decline observed between 2006 - 2010 may represent the initial re

of the U.S. EPA’s 2010/2015 PFOA Stewardship Program.
Epidemiology Associations Reporte

with PFOS and PFOA

Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion. The measurements we obtain may afford only a 

byproduct of the underlying pharmacokinetics, systems, biology, and pathogthese are not entirely new concepts, their importance may be clearer today before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion. The measurements we obtain may afford only a byproduct of the underlying pharmacokinetics, systems, biology, and pathogthese are not entirely new concepts, their importance may be clearer today before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion. The measurements we obtain may afford only a byproduct of the underlying pharmacokinetics, systems, biology, and pathogthese are not entirely new concepts, their importance may be clearer today before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion. The measurements we obtain may afford only a byproduct of the underlying pharmacokinetics, systems, biology, and pathogthese are not entirely new concepts, their importance may be clearer today before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance today than ever before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance 

today than ever before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance 

today than ever before.”

Epidemiologic associations are not equivalent to causation.
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance 

today than ever before.”

Epidemiologic associations are not equivalent to causation.

May expect physiologicaassociations
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance 

today than ever before.”

Epidemiologic associations are not equivalent to causation..

May expect physiologicaassociations“Reverse ca
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance 

today than ever before.”

Epidemiologic associations are not equivalent to causation.

May expect physiologicaassociations“Reverse ca
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance today than ever before.”

Epidemiologic associations are not equivalent to causation.

May expecphysiologicassociation“Reverse c
Is Clearance a Common Explanation Across the Epidemiological A

Reported at “Low” PFOA and PFOS Concentrations?
Specifically, Is Clearance a Common Explanation Across These Reported Epidemiologic
Definition of Clearance

1. the act of clearing.2. a quantitative measure of the rate at which a substance is removed from the blood, as by the kidneys, the liver, gut.3. expressed as the volume of plasma cleared per unit time.Cl = Kel x Vd
Study Populations and Metho

Study Populations

• General population

NHANESDenmarkOther

• Community

Parkersburg, W.V. (mid-Ohio River)

C8 Health Project 

Epidemiologic Study De

• Cross-sectional

General populatioCommunityOccupational

Medical survei

• Case-control

General populatio

Longitudinal (cohor

C8 Health Project C8 Science Panel

• Occupational

3M Company

Decatur, ALCottage Grove, MNAntwerp, Belgium

DuPont

Washington Works (W.V.)

Miteni (Italy)

• Longitudinal (cohor

CommunityOccupational

• Case-cohort• Cohort mortality

CommunityOccupational

Study Populations and Metho

Study Populations

• General population

NHANESDenmarkOther

• Community

Parkersburg, W.V. (mid-Ohio River)

C8 Health Project 

Epidemiologic Study De

• Cross-sectional

General populatioCommunityOccupational

Medical survei

• Case-control

General populatio

Longitudinal (cohor

C8 Health Project C8 Science Panel

• Occupational

3M Company

Decatur, ALCottage Grove, MNAntwerp, Belgium

DuPont

Washington Works (W.V.)

Miteni (Italy)

• Longitudinal (cohor

CommunityOccupational

• Case-cohort• Cohort mortality

CommunityOccupational
Approximate Average PFOA Concentrations (ng/m

3000400050006000700080009000

10000

Concentration (ng/mL)

5 15 28 67 224 500 650

1500

1000

0

10002000

3000

General Population Washington County, MN Other W. Va Water Districts Lubeck water district

Little Hocking Water Association DuPont Washington Works 3M Antwerp 3M Decatur 3M Cottage Grove (ECF cells) 3M Cottage Grove (spray dr

Concentration (ng/mL)
Approximate Average PFOS Concentrations (ng/mL

300

300400500600700800900

1000

Concentration (ng/mL)

20 35 20 20 20 20 50

0

100200

300

General Population Washington County, MN Other W. VaLubeck

Little Hocking Water Association DuPont Washington Works 3M Cottage Grove 3M Cottage Grove (work-up) 3M Antwerp 

Concentration (ng/mL)
Comparison of Human PFOA Biomonitoring Data t

Benchmark Lower Bound Internal Concentrations from Tox

Study Endpoints

400006000080000

100000120000140000

Concentration (ng/mL)Human Biomonitoring Toxicolo

5 15 28 67 224 500 650 1500 1000 5000 5600

19000230

0

20000

40000

General Population

Washington County, MN

Other W. Va Water Districts

Lubeck water district

Little Hocking Water Association

DuPont Washington Works

3M Antwerp

3M Decatur

3M Cottage Grove (ECF cells)

3M Cottage Grove (spray dryer)

Miteni (Italy)

Post-natal survival at weaning (mouse)

Liver weight-to-brain-weight ratio (monkeys)

Post-weaning mortality 

Body weight chan

Tuo

Concentration (ng/mL)
Comparison of Human PFOS Biomonitoring Data t

Benchmark Lower Bound Internal Concentrations from To

Study Endpoints

16,000

31,000

44,000

4

200003000040000500006000070000

Concentration (ng/mL) Human Biomonitoring

Toxicology

20 35 20 20 20 20 50 300 900 900

16,000

0

1000020000

General Population

Washington County, MN

Other W. Va

Lubeck

Little Hocking Water Association

DuPont Washington Works

3M Cottage Grove

3M Cottage Grove (work-up)

3M Antwerp

3M Decatur

Postnatal survival to day 8 (rats)

Pup weight gain through weaning (rats)

Liver weight (male rats)

Liver weight (female monkeys)

Postnatal survival to day 6 (

Liver tum

Concentration (ng/mL)
Is Clearance a Common Explanation Across the Reported Epidemiological 
Delayed Puberty

Lopez-Espinosa et al. Environ Sci Technol 2011;45:8160-8166

C8 Health ProjectCross Sectional Study3076 boys2931 girls8 – 18 years of age
Boys Quartile Geometric Mean (GSD)
Girls
Lopez-Espinosa et al. Conclusion:

“Results suggest later age at having reached puberty of 3 to 6 months acro

concentrations found in this population for both boys and girls.”
Considerations re Delayed Puberty Association

For those who had reached puberty, the measure of serum perfluoroalkyls after puberty had been reached.Physiological changes may explain the associations reported

MenstruationExpanding muscle/tissue mass
Time to Pregnancy

(subfecundity

)

Definition: > 12 months to become pregnant
Odds Ratio for Subfecundity (TTP > 12 months). 

Fei et al. Human Repro 2009;24:1200-1205

Study Pop: DenmarkStudy Design: CohortN = 1240

PFOS < 26.0 ng/mLPFOA < 3.91 ng/mL PFOS 26. 1 – 33.3 

ng/mLPFOA 3.91 – 5.20 ng/mL

PFOS 33.4 –43.2 ng/mLPFOA 5.21 –6.96 ng/mL

PFOS ≥ 43.3 ng/mLPFOA ≥ 6.97 ng/mL
Suggested causal diagram of maternal perfluorochemical concentrations and time to 

Olsen GW, Butenhoff JL, Zobel LR. Repro Toxicol 2009.
Acknowledgement: Dr. Matt Longnecker (NIEHS). Presented at EPA PFAA Days III (June 2010). See Whitworth et al. Ep
Odds Ratio for Subfecundity (TTP > 12 months). 

Whitworth et al. Epidemiology 2012;23:257-263

Study Pop:NorwayStudy Design: Case-ControlN = 421 cases, 504 controls

PFOS < 10.34 ng/mLPFOA < 1.66 ng/mL PFOS 10.34 – 13.09 ng/mL

PFOA 1.66 – 2.24 ng/mL

PFOS 13.10 –16.60 ng/mLPFOA 2.25 –3.02 ng/mL

PFOS ≥ 16.61 ng/mLPFOA ≥ 3.03 ng/mL
Odds RatioO Subfecundity (TTP > 12 months) Stratified by Par

Whitworth et al. Epidemiology 2012;23:257-263 Study Pop: Norway

Study Design: Case-controlN: 421 cases, 504 controls
Olsen, Butenhoff, Zobel Repro Toxicol 2009

Fei et al. Epidemiology 2012

“A subtlety not captured by the directed acyclic graph is that PFOA/PFOS were not measured at th

of the attempt at conception(which would have been ideal), but at the end, after a pregnancy had 

Thus, in the available data, the measurement of PFOA/PFOS can potentially be influenced by TTP 

through reaccumulation of the chemicals. Such influence produces a cycle in the graph through t

to the measured PFOA/PFOS. However, for nulliparous women, that arrow does not exist in a mo

adjusts for age.”
Olsen, Butenhoff, Zobel Repro Toxicol 2009

Fei et al. Epidemiology 2012

parous women

“A subtlety not captured by the directed acyclic graph is that PFOA/PFOS were not measured at th

of the attempt at conception(which would have been ideal), but at the end, after a pregnancy had 

Thus, in the available data, the measurement of PFOA/PFOS can potentially be influenced by TTP 

through reaccumulation of the chemicals. Such influence produces a cycle in the graph through t

to the measured PFOA/PFOS. However, for nulliparous women, that arrow does not exist in a mo

adjusts for age.”

parous women
Odds Ratio for Subfecundity (TTP > 12 months) Stratified by Pa

Fei et al. Epidemiology 2012:23:264-266

Study Pop: DenmarkStudy Design: CohortN = 1240

PFOS Quartile
Conclusions

Time to Pregnancy 

Whitworth et al. “Among parous women, increased body burden may be duinterpregnancy interval rather than the cause of a long time to pregnancy. T

from nulliparous women may be more informative regarding toxic effects of 

perfluorinated compounds. Our results among nulliparous women did not sassociation with subfecundity.”Fei et al. “We found limited evidence for reverse causation as an explanation 
Lower Birthweight
Author Type Journal Year Loc

Olsen et al. Occupational JOEM 2004 3M DGrice et al. Occupational JOEM 2007 3M DApelberg et al. (2 papers)

General population

ES&T and EHP 2006, 2007 Baltim

Fei et al. (4 papers)

General population

EHP (2), Am J Epi (1), Human Repro (1)

2007, 2008, 2008, 2009

Denm

Inoue et al. General 

population

EHP 2004 Hoik

Monroy et al. General 

population

EnvironmentalResearch

2008 Onta

Nolan et al. (2 papers)

Community Reproductive 

Toxicology

2008, 2009 Was

CounHeal

Stein et al. Community Am J Epidemiology 2009 “6 wa

C8 H

Hamm et al. General 

population

J Exp Sci Env Epi 2009 Edm
Covariates included in statistical models of birth weig

PFOS or PFOA, by study investigator

Adjusted For:

Olsen Grice Inoue Apelberg Fei Monroy Washino No

Maternal age Yes Yes Yes Yes Yes YeMaternal BMI Yes Yes Yes Yes Maternal net wt gain

Yes

Maternal height Yes Maternal race Yes YeMaternal smoking

Yes Yes Yes Yes Yes 

Education Yes SES Yes YeGestational age Yes Yes Yes Yes YeGetational age at blood draw

Yes Yes

Parity Yes Yes Yes Yes Yes Infant sex Yes Yes Yes Yes YeDiabetes YesHypertension Yes
Endpoint PFC Olsen Grice Inoue Apelberg Fei 

(4 papers)

Monroy Washino Nolan

(2 pa

Gestational age PFOS N.S. N.S. N.S. 

PFOA N.S. N.S. N.S. N.S. 

Birth Weight (g) PFOS N.S. N.S. N.S.* N.S. N.S. S.S.females 

PFOA N.S N.S.* S.S. N.S. N.S. N.S. 

Birth Length (cm) PFOS N.S. N.S. N.S.

PFOA N.S. S.S. N.S.

Head Circumference (cm)

PFOS S.S. N.S. N.S.PFOA S.S. N.S. N.S.

Abd/chest Circumference (cm)

PFOS N.S. N.S.PFOA S.S. N.S.

Apgar Score PFOS N.S.

PFOA N.S.

Ponderal Index PFOS S.S. N.S.

PFOA S.S. N.S.

Placental Weight (g) PFOS N.S.

PFOA N.S.

Miscarriage PFOS N.S. N.S. 

PFOA N.S. N.S. 

Birth defects (nonspecific) PFOS N.S. 

PFOA N.S. N.S. 

Preeclampsia PFOS 

PFOA 

Developmental milestones

PFOS N.S.PFOA N.S.
C8 Science Panel Very Recent Publications

Savitz et al. Epidemiology 2012 doi:L10.1097/EDE.0b013e31824cb93Savitz et al. Environ Health Perspect 2012 doi:org/10.1289/ehp.11047
C8 Health Project enrollees (Savitz et al. Epidemiology 20

Self-reported pregnancy histories form the C8 Health Project partiReconstructed historical exposures for C8 Health Project participabeginning in 1951.Use modeled estimates of PFOA exposure at the time of pregnancPFOA and pregnancy outcomes within this C8 Health Project popu1990 – 2006 time frame; 11,737 pregnancies1) Adjusted OR (95% CI) for IQR (lnPFOA)increase where = 2.192) Quintiles (2 lowest quintiles used as reference)3) Adjusted OR (95% CI) for 100 ng/mL increase in PFOA

OR 95% CI

Miscarriage 0.98 0.93 – 1.03Stillbirth 0.91 0.76 – 1.10Preeclampsia 1.08 1.01 – 1.15Preterm Birth 0.97 0.93 – 1.02Term Low Birthweight 0.96 0.79 – 1.16Birth Defect 0.97 0.90 – 1.06

Conclusion: “No associations between estimated serum PFOA levepregnancy other than possibly preeclampsia. 
Savitz et al. Environ Health Perspect 2012

TWO StudiesSTUDY 1. 

Examine birth certificate (OH and WV) and fetal death records (Wcounties (3 Ohio, 2 WV)Restricted to pregnancies from 1990 – 200410% random sampling of term births ≥2500 grams (n = 3616) sel

geocoding for PFOA exposure

PFOA exposure history considered at time of pregnancy and assulived at residence for 6 years prior to pregnancy1) Adjusted OR (95% CI) for IQR (lnPFOA) increase where = 2.192) Quintiles (2 lowest quintiles used as reference)3) Adjusted OR (95% CI) for 100 ng/mL increase in PFOA
STUDY 1 (change in 100 ng/mL)

OR 95% CI

Stillbirth 1.20 0.86 – 1.68Pregnancy-induced 

hypertension 1.06 0.86 – 1.31

Preterm births

<37 weeks gestation 1.02 0.94 – 1.10<32 weeks gestation 0.90 0.74 – 1.10

Term low birthweight 1.00 0.86 – 1.15Term SGA 0.96 0.67 – 1.11Term birthweight

(Change in Birthweight, g) -14.8 -43.28, 13.68

(By Quintiles of PFOA Exposure) OR 95% CI

Q1-Q2 (<6.1 ng/mL) referenceQ3 (6.1 - < 10.2 ng/mL) 22.8 -32.9, 78.5Q4 (10.2 – 21.0 ng/mL) 2.3 -50.3, 54.8Q5 (21.0 – 717.6 ng/mL) -9.5 -58.4, 39.4
STUDY 2

Linked birth records from 1990 – 2004 for 13 counties (5 Ohio, 8 WV) to sfrom the C8 Health Project (5 of these counties)Linked (matched) 4547 birth certificates linked to the C8 Health ProjectUsed lifetime residential history to calculate annual exposure PFOA estim
STUDY 2 (change in 100 ng/ml)

OR 95% CI

Pregnancy-induced 

hypertension 1.01 0.91 – 1.12

Preterm births

<37 weeks gestation 1.09 1.00 – 1.18<32 weeks gestation 1.10 0.86 – 1.40

Term low birthweight 1.00 0.82 – 1.21Term SGA 1.07 0.98 – 1.17Term birthweight

(Change in Birthweight, g) -9.14g -20.30, 2.02

(By Quintiles of PFOA Exposure) OR 95% CI

Q1-Q2 (<8.9 ng/mL) referenceQ3 (8.9 - < 21.8 ng/mL) -3.8 -40.4, 32.8Q4 (21.8 – 83.3ng/mL) -25.4 -63.7, 12.9Q5 (83.3 – 921.3 ng/mL) -33.3 -73.1, 6.5
Conclusion Savitz et al. (Environ Health Perspect)

Little support for PFOA having adverse effects on pregnancy-

hypertension or preterm birthInconsistent evidence of reduced fetal growthImprecise indications of an association with early preterm biThe potential association of PFOA with small decrements in bunknown significance, but is perhaps the most readily amenarefined studies.
Conclusion (continued)

Savitz et al. Environ Health Perspect, 2012“If an impact on birth weight is causal, which is yet to b

it reflects a biological process that is worth elucidating 

of direct health concern.”
Considerations(Olsen et al. Reprod Toxicol 2009;27:212-230)

• PFOA and PFOS distributed primarily in extracellular fluid•Protein bound• Increased maternal plasma volume expansion and placental volum

are strongly correlated with birth weight.

Other referencePlasma Volume Expansion in Pregnancy: Implications for Biomarkers in Population Stud

Faupel-Badger et al. Cancer Epidemiology Biomarkers & Prevention 2007:16:17201725.
Duration of Breastfeeding

(Fei et al. Scand J Work Environ Health 2010;36:413-421)

DenmarkCohort study1400 pregnant women
Higher maternal PFOS and PFOA concentrations were associated with short

of breastfeeding. For example, median PFOS concentration was 37.0 ng/mwho breast fed < 3 months compared to 32.3 ng/mL for women who breasFor PFOA, these values were 5.90 ng/mL and 4.96 ng/mL, respectively.

The duration of breastfeeding decreased with increasing concentrations of p

PFOS and PFOA among multiparous women but not among primiparous wo
Fei et al. conclusion. “These findings suggest that PFOA and PFOS may re

ability to lactate, but could equally reflect reverse causation since no asswas seen in primiparous women.”
Menopause

Knox et al. Implications of early menopause in women exposed to perfluorocarbons. J ClinEndocrinol Metab 2011;96:1747-1753

C8 Health ProjectCross Sectional Study25,957 women aged 18 - 65
Menopause is the permanent end of

menstruation and fertility, defined asoccurring 12 months after the lastmenstrual period. Menopause in the Knox et al analysis was defined by the participant’s answer to the question of whether she had experienced the “change of life” (menopause).They were not asked the age of menopause.
TABLE 2. Odds ratios for menopause by quintile of PFOA/PFOS

 Women 18 ≤ 42 yr Women > 42 ≤ 51 yr Women > 51 ≤ 65 

 Point Confidence Point Confidence Point Confide estimate limits Χ2 estimate limits Χ2 estimate limits PFOAa

PFOS quintiles.Quintiles in ng/mL. Quintile 1Quintile 2: 11.9 - 17.0, QuintiQuintile 4: 22.5 - 30.7, Quinti

PFOA quintiles.Quintiles in ng/mL. Quintile 1: 0.25 - 11.2, Quintile 2: 11.3 - 19.8, Quintile 3: 19.9 - 36.7, Quintile 4: 36.8 - 84.9, Quintile 5: 85.0 - 22412.0.

PFOAa2 vs, 1 0.9 0.5 – 1.6 0.009 1.4 1.1 – 1.8 0.839 1.5 1.1 – 2.3 vs. 1 0.9 0.5 – 1.5 1.2 0.9 – 1.6 1.6 1.2 – 2.4 vs. 1 0.9 0.5 – 1.7 1.4 1.1 – 1.9 1.4 1.1 – 1.5 vs. 1 1.2 0.7 – 2.1 1.4 1.1 – 1.8 1.7 1.3 – 2.PFOSa

2 vs, 1 1.1 0.7 – 1.9 0.804 1.2 0.9 – 1.5 0.028 1.5 1.1 – 2.

3 vs. 1 0.8 0.4 – 1.4 1.4 1.1 – 1.8 1.8 1.3 – 2.4 vs. 1 1.0 0.6 – 1.8 1.4 1.1 – 1.8 2.0 1.5 – 2.5 vs. 1 1.1 0.6 – 2.1 1.4 1.1 – 1.8 2.1 1.6 – 2.Results are adjusted for age, BMI, alcohol consumption, smoking, and exercise. Women with hysterectomy were excluded a. Quintiles. 
“Mean PFC concentrations in women with and without hysterectomy 

compared in women 40-55 yrs of age to see whether hysterectomy 

difference. Both PFOS and PFOA were significantly higher (P < 0.000

with a hysterectomy.”

Knox et al. J Clin Endocrinol Metab 2011
“Thus, the argument could be made that some of the significance asso

increased risk of menopause associated with increased PFC might be dcausation, i.e., the disappearance of menses causing increasing PFC co

rather than an increase in PFC causing early menopause.

Knox et al. J Clin Endocrinol Metab 2011

This argument is given additional support by the fact that women who hysterectomy in our sample had higher mean levels of PFC than womenot experienced hysterectomy.”
“However even if the results reported here could be solely explained b

causation, they would still be cause for alarm because it would imply tPFC exposure is the natural result of menopause or hysterectomy.”

Knox et al. J Clin Endocrinol Metab 2011
Is Clearance a Common Explanation Across the Reported Epidemiological 
Chronic Kidney Disease (CKD

Estimate of Net Renal Tubular Secrtion or Reabsorption o

in Different Species (Han et al. Chem Res Toxicol 2012;25:35-46)

Species Gender GFR (L/d/kg) CLR

(mL/d/kg)

Net secretion (mL/d/kg)

Net reabosprtion(mL/d/kg)

Rabbit Female 4 670 590 

Male 4 640 560 

Rat Female 14.4 666 378 Dog Female 5.3 50.8 55 

Male 5.3 43 63 

J.macaque Female 8.5 32 138 

Male 8.5 15 155 

Rat Male 14.4 18.2 270 Mouse Female 16.7 16 318 

Male 16.7 10 324 

Human 2.57 0.03 51 
glomerular filtration rate (GFR) an expression of the quantity of glomeru

formed each minute in the nephrons of both kidneys, usually measured bof clearance of creatinine . Ideal measures of GFR includes use (infusionas it is freely filtered but not secreted or reabsorbed in tubules.Creatinine-based approximations of GFR (eGFR)

Filtered freely by the glomerulus, but also actively secreted by tperitubular capillaries in very small amounts such that creatininclearance overestimates GFR by 10 to 20 percent. Creatinine levels affected by muscle mass, ingetion of meats1) 24-hour urine collection to measure creatinine2) Several indirect formulas

Cockgroft-GaultCKD-EpiMDRD
Modification of Diet in Renal Disease (MDRD) formula

estimated Glomerular Filtration Rate (eGFR) =175 x (serum creatinine in mg/dL)-1.154 x (age in years)-0.203 x (0.742 if female) x 

Chronic Kidney Disease (CKD) defined as eGFR < 60mL/min
Study Pop: NHANES 1999-2000, 2003-2004, 2005-2006

Study Design: Cross-sectional

Association Between Plasma Perfluoroalkyl Level and eGFR

Shankar et al. Am J Epidemiol 2011;174:893-900
Shankar et al. conclusions:

•Results suggest that elevated PFC levels are associated with CKD

• Cross-sectional study does not address temporality.• In CKD, when eGFR is reduced, the total filtration of PFOA and PFO

reduced. This could lead to higher concentrations of PFOA and PFO

result in a positive association between these perfluoroalkyls and C

This is referred to as reverse causation.• An argument against ‘reverse causality’ is that the inverse associati

among persons without CKD.

• A ‘reverse causality’ association could be of clinical and/or public h

because if subjects with CKD retain these compounds, the toxicities 

PFOS on the kidneys or other organs could be accentuated among C
80

100120140

80

100120140

Study Pop Scatterplot of eGFR by PFOA and PFOS Serum Concentratio

Olsen and Zobel Int Arch Occup Environ Med 2007;81:231-246

Study Pop: OccupationalStudy Design: Cross-sectional

0

20406080

1 10 100 1000 10000 100000

eGFR

PFOA (ng/mL)

0

20406080

1 10 100 

PFOS (ng/mL
Study Pop: Occupational

Study Design: Cross-sectional

PFOA and PFOS Serum Concentrations by Deciles

Olsen and Zobel Int Arch Occup Environ Med 2007;81:231-246
Adjusted Mean eGFR by PFOA and PFOS Deciles

Olsen and Zobel Int Arch Occup Environ Med 2007;81:231-246

Study Pop: OccupationalStudy Design: Cross-sectional
Adjusted Odds Ratio for Chronic Kidney Disease by PFOA and PFOS 

Olsen and Zobel Int Arch Occup Environ Med 2007;81:231-246

Study Pop: OccupationalStudy Design: Cross-sectional
Chronic Kidney Disease (eGFR) Results from C8 Heal
Chronic Kidney Disease (eGFR) Results from C8 Healt

Never Reported
C8 Health Project

Association Between C8 Deciles and Creatinine, Serum (m

S ifi d b A & G d

C8 Health Project

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/pdfs/Cre
C8 Health Project

Association Between C8 Deciles and Albumin, Serum (g‐

S ifi d b A & G d

C8 Health Project

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/pdfs/Album
NHANES 

2009 

- 2010
NHANES 2009 – 2010

eGFR by ln PFOA eGFR by ln PFOS
CONCLUSIONS regarding Chronic Kidney Disease (CKD)

No evidence of CKD via medical surveillance where workerconcentrations of PFOS and PFOA much higher than NHANNo evidence toxicologically.NHANES results by Shankar et al likely an association of ren(eGFR and PFOA/PFOS) and NOT CKDNHANES 2009-2010 results continue to show eGFR associatlower levels of PFOA/PFOS than reported by Shankar et al.eGFR and PFOA/PFOS are positively associated which is bioplausible .
Inference: Health outcomes associated with eG

could therefore also be found to be associated 

concentrations of PFOA/PFOS.
Hyperuricemia

Steenland et al. Environ Health Perspect 2010;118:229Shankar et al. Clinical Epidemiol 2011;3:251-258

NHANES 1999-2000 and 2003-2006 (Shankar et al.)C8 Health Project (Steenland et al.)
Shankar et al. (NHANES, Cross-sectional)
Steenland et al. (C8 Health Project, Cross-sectional)
Is uric acid associated with eGFR

?
NHANES 2009 - 2010

eGFR by Uric Acid eGFR by ln Uric Acid
eGFR and Systolic BP Systolic BP and ln PFOA Systolic BP 

NHANES 2009 - 2010
Steenland et al.

“Only 119 people (0.2%) had c>2.5 mg/dL, which suggests kidthat can affect uric acid; excludparticipants from the analyses the results. A broader exclusio1.5 mg/dL excluded 739 and ageffect on results.”“We also analyzed a subset of had a PFOA level < 20 ng/mL, dquartiles of PFOA exposure, so group had levels similar to thogeneral population. Those wit(n = 2,012), 10-14.9 ng/mL (n= 15-20 ng/mL (n=7,387) had prein uric acid of 0.14, 0.21, and 0above those with PFOA levels < 
C-reactive Protein
C8 Health Project

Association Between C8 Deciles and C‐Reactive Protein, Q

( L)

C8 Health Project

(mg‐L)

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/p
C8 Health Project

Association Between C8 Deciles and Creatinine, Serum (m

S ifi d b A & G d

C8 Health Project

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/pdf
Diabetes
1.0

1.52.0

O

dd

s

R

a

Odds Ratio for Medically Verified Type II Diabetes, C8 HeMacNeil et al. Environ Res 2009;109:997-1003

0.00.5

Decile 1 

(<7.9 

ng/mL)

Decile 2 

(8.0 - 11.6 

ng/mL)

Decile 3 

(11.7 -

15.6 

ng/mL)

Decile 4 

(15.7 -

20.8 

ng/mL)

Decile 5 

(20.9 -

28.0 

ng/mL)

Decile 6 

(28.1 -

39.4 

ng/mL)

Decile 7 

(29.5 -

57.5 

ng/mL)

Decile 8 

(57.6 -

89.7 

ng/mL)

a

t

i

o

PFOA Decile
C8 Health Project

Association Between C8 Deciles and Glucose, Serum ( mg‐d

S ifi d b A & G d

C8 Health Project

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/pd
C8 Health Project

Association Between C8 Deciles and Creatinine, Serum ( m

S ifi d b A & G d

C8 Health Project

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/pdf
C8 Health Project

Association Between C8 Deciles and Albumin, Serum (g‐

S ifi d b A & G d

C8 Health Project

Stratified by Age & Gender

Original slide containing figure in presentation was removed

due to copyright; however, image can be viewed at the

following link:

http://www.hsc.wvu.edu/som/cmed/c8/results/otherLaboratoryValues/pdfs/Album
MacNeil et al. Conclusion

“Our findings do not demonstrate abn association between PFOA and eOr fasting glucose level. Our data are limited by tehir cross-sectional naNot preculde the possibility of a causal relationship.”
What does an increased risk for diabetes (type II) 

in the lowest PFOA decile

of the C8 Health Project database mean 

when other health conditions are cross-sectiona

with respect to concentrations of PFOA 
Occupational Cohort Mortality Studies

Diabetes

Study Categorization Obs Exp SMR Alexander et al. Decatur Facility 

(AL comparison)

0 1.5 0.0 

Lundin et al. Cottage Grove Facility 

(MN comparison)

23 20.0 1.2 

“Not exposed PFOA job” “Not exposed PFOA job”

5 9.6 0.5 

Ever “Probable exposure” 18 8.9 2.0 Ever “Definite exposure” 0 1.5 0.0 

Leonard et al. Washington Works Facility 

(W.V. comparison)

22 31.5 0.7 

Washington Works Facility (DuPont comparison)

22 11.2 2.0 

Is Clearance a Common Explanation Across Many of these Epidemiologica
High Cholesterol
PFOA and the Linear Associations (Slope) Between Cholesterol and PFOA in 10 Epide

Steenland et al. Environ Health Perspect 2009;118:1100-1108 and Olsen et al. J Occup Environ Med 20
Nonlinear Association Between Cholesterol and PFOA and PFOS

Steenland et al. Am J Epidemiology 2009;170:1268-78

Study pop: C8 Health ProjectStudy Design: Cross-sectional

Note: Scale ofX axis for PFOS
Nonlinear Association Between Cholesterol and PFOA and PFOS

Steenland et al. Am J Epidemiology 2009;170:1268-78

Study pop: C8 Health ProjectStudy Design: Cross-sectional

Note: Scale ofX axis for PFOS
Adjusted Mean Lipid Values by PFOA Deciles 

Olsen and Zobel Int Arch Occup Environ Health 2007;81:231-246

150200250

Lipid value

Study pop: 3M occupationalStudy Design: Cross-sectionalN = 506 male employees not taking cholesterol lowering medications

0

50

100

Decile 1

median 60

range 7-130

Decile 2

median 190

range 131-290

Decile 3

median 360

range300-440

Decile 4

median 540

range 450-710

Decile 5

median 910

range 720-

1110

Decile 6

median 1250

range 1120-

1400

Decile 7

median 1630

range 1420-

1850

Decile 8

median 2180

range 1860-

2500

Decile 9

median 2960

range 2510-

3690

Decile Number ( PFOA median and range in ppb for each decile)

Lipid value

ChLDHDTr
Olsen et al. Longitudinal assessment of lipid and hepatic clinical parameters i

Involved with the demolition of PFOA and PFOS manufacturing facilities. JOEM 
-120

-100

-80-60-40-20

0

20406080

100120

-3 -2 -1 0 1 2 3 4 5 6 7

Change in Non-HDL mg/dL

Change in ln PFOA ng/mL

-120-100

-80-60-40-20

0

20406080

100120

-3 -2 -1 0 1 

Change in Non-HDL mg/dL

Change in ln PFO

-60-40-20

0

204060

-3 -2 -1 0 1 2 3 4 5 6 7

Change in HDL mg/dL

Change in ln PFOA ng/mL

-60-40-20

0

204060

-3 -2 -1 0 1 

Change in HDL mg/dL

Change in ln PFO
Steenland et al. Am J

Epidemiol 2009; 170:1268-78

Study pop: C8 Health ProjectStudy Design: Cross-sectional

Nonlinear Association Between PFOA, Total Cholesterol and U

Steenland et al. Environ Health Perspect 2010;118:229-233
Study pop: C8 Health Project

Study Design: Cross-sectional

Steenland et al. Am JEpidemiol 2009; 170:1268-78

Nonlinear Association Between PFOS, Total Cholesterol and Uric Ac

Steenland et al. Environ Health Perspect 2010;118:229-233
Is nonHDL cholesterol associated with the glomerular filt
Morita et al. Atherosclerosis 2010. Decrease in glomerular filtration rat

low-density lipoprotein cholesterol in subjects with normal kidney functiby urinalysis and plasma creatinine . Atherosclerosis 2010;210:602-606.

Subjects were examinees at a wellness and promotion program in JapaPatient levels < 1 mg/ Patient levels < 1 mg/dL for men and 0.8 mg/ for men and 0.8 mg/dL for females w/o for females w/o album

album

4602 males; 2920 females.Cross-sectional analysis
Morita et al. Atherosclerosis 2010. Decrease in glomerular filtration rate by 

plasma low-density lipoprotein cholesterol in subjects with normal kidney function assessed by urinalysis and plasma creatinine . Athersclerosis2010;210:602-606.
Morita et al. Atherosclerosis 2010
Morita et al. Atherosclerosis 2010;210:602-606

Conclusion: “We conclude that the high plasma level of LDL-C was t

factor among coronary risk factors to reduce GFR probably due to imfunction of renal arterioles and capillaries of glomeruli in subjects w

function assessed by urinalysis and plasma creatinine.”
Lo et al. GFR, body mass index, and low high-density lipoprotein con

adults with and without CKD. Am J Kidney Dis 2007:50:552-558.

Cross sectional analysis of 380,207 individuals from Kaiser Permanente who outpatient measured of serum creatinine from 2003 – 2004. 

Conclusion: Decreased eGFR is independently associated with greater oddhaving a low HDL level (men < 40 mg/dl; females < 50 ng/mL).

“Thyroid Disorders”

Knox et al. J Toxicol Sci 2011Melzer et al Environ Health Perspect 2010Lopez-Espinosa et al. Environ Health Perspect 2012C8 Science Panel. Status Report 2011.Olsen and Olsen and Zobe. l Int Arch Occup Environ Health 2007 Environ Health 2007

NHANESC8 Health Project

(adults and children)

OccupationalAll cross-sectional studies
Melzer et al. Environ Health Perspect 2010;

NHANES Data 1999-2000, 2003-2004 and 2005-2006Cross-sectional
Knox et al. J Toxicol Sci 2011;36:403-410

C8 Health ProjectCross-sectionalN = 52,296

No association with TSH

Lopez-Espinosa et al. Environ Health Perspect 2012

C8 Health ProjectCross-sectional10,725 children (aged 1 – 17 years)
Lopez-Espinosa et al. Environ Health Perspect 2012
1.0

1.52.02.53.0

O

dd

s

R

a

t

i

WomenMen

C8 Science Panel Status Report 2011. Odds Ratios for Thyroid Disorders an

C8 Health Project

0.00.5

Decile 1 (< 42 ng/mL)

Decile 2 (42 - 93 ng/mL)

Decile 3 (94 - 127 ng/mL)

Decile 4 (128 - 167 ng/mL)

Decile 5 (168 - 238 ng/mL)

Decile 6 (238 - 400 ng/mL)

Decile 7 (401 - 821 ng/mL)

Decile 8 (822 - 2,226 ng/mL)

Decile 9 (2,227 - 6,028 ng/mL)

i

o

Cumulative PFOA Decile
Adjusted Mean Thyroid Values by PFOA Deciles 

Olsen and Zobel (Int Arch Occup Environ Health 2007)

5.06.07.08.09.0

10.0

Value of Thyroid Test

OccupationalCross-sectional

0.01.02.03.04.05.0

Decile 1

median 60

range 7-130

Decile 2

median 190

range131-290

Decile 3

median 360

range300-440

Decile 4

median 540

range450-710

Decile 5

median 910

range 720-

1110

Decile 6

median 1250

range 1120-

1400

Decile 7

median 1630

range 1420-

1850

Decile 8

median 2180

range 1860-

2500

Decile 

median 2

range 25

3690

Decile Number (median and range PFOA in ppb for each decile)

Value of Thyroid Test 
Are thyroid diseases related to eGFR?
Åsvold et al. Association of thyroid function with estimate

glomerular filtration rate in a population-based study: the Eur J Endocrinol 2011;164:101-105Cross-sectional, Norwegian population based study of 29,480 individuals abovof age without previously known thyroid disease. Calculated a geometric mean eGFR and odds ratio of chronic kidney disease (60 ml/min per 1.73 m

2) according to categories of thyroid function, using peo

TSH in the lower third of the reference range (0.50 – 1.4mU/L) as the compar
Åsvold et al., Eur J Endocrinol 2011.
Åsvold et al., Eur J Endocrinol 2011 
Lo et al. Increased prevalence of subclinical and clinical hypothyroidism in p

chronic kidney disease. Kidney International 2005;67:1047-1052

NHANES 1988 – 1994N 14,623Cross-sectional
Prevalence of hypothyroidism by level of estimated glomerular filtration 

0.0001 for trend.

Lo et al. . Increased prevalence of subclinical and clinical hypothyroidism in chronic kidney disease. Kidney International 2005

NHANES 1988 – 1994N 14,623Cross-sectional
Lee-O!

eGFR Cardiac Output Resistance 

Vascular

SUMMARYHypothyroidism, Hyperthyroidism, and eGFR

HypothyroidismHyperthyroidism
eGFR Cardiac Output Resistance Blood F

Arteriole Vascula

SUMMARYHypothyroidism, Hyperthyroidism, and eGFR

HypothyroidismHyperthyroidism
“To the best of our knowledge, this is the first large-scale report of an assoc

PFAAs in serum and thyroid function impairment in children aged 1 – 17 yeaof 10,725 children from the Mid-Ohio Valley, measured serum PFOA concen2005 were positively associated with thyroid disease (mostly hypothyroidismSerum PFOS and PFNA concentrations were associated with slightly higher lin children 1 – 17 years, but were not positively associated with subclinicalhyperthyroidism.”

Lopez-Espinosa et al. Environ Health Perspect 2012
Hepatic Enzymes (ALT)

Lin et al. Am J Gastroenterology 2009Gallo et al. Environ Health Perspect 2012Olsen and Zobel 2007 

NHANESC8 Health ProjectOccupationalCross-sectional
20

25303540

PFOA Quartiles

ALTGGT

2025303540

PFOS Quart

Lin et al. Unadjusted Geometric Mean of ALT and GGT (NHANES, Cross-secti

05

1015

Q1 (<2.91 

ng/mL)

Q2 (2.91 -

4.2 ng/mL)

Q3 4.21 -

5.95 ng/mL)

Q4 (>5.95 

ng/mL)

05

1015

Q1 (<15.51 

ng/mL)

Q2 (15.51 - 23.50 

ng/mL)

Q3 (

3

ng
Gallo et al. (C8 Health Project, Cross-sectional)

Fitted values of ALT from fully adjusted regression models by deciles of PFOS the mean values of the other covariates
Adjusted Mean Liver Clinical Chemistry Values by PFOA Deciles 

Olsen and Zobel (Int Arch Occup Environ Health 2007)

5060708090

Value of Liver Test

AlASALG

Occupational, Cross-sectional

0

10203040

Decile 1

median 60

range 7-130

Decile 2

median 190

range131-290

Decile 3

median 360

range300-440

Decile 4

median 540

range450-710

Decile 5

median 910

range 720-

1110

Decile 6

median 1250

range 1120-

1400

Decile 7

median 1630

range 1420-

1850

Decile 8

median 2180

range 1860-

2500

D

me

ran

Decile Number (median and range PFOA in ppb for each decile)

Value of Liver Test
Lin et al. (NHANES data): “In conclusion, using the NHANES data from the US 

population, we found that a higher serum concentration of PFOA was associatElevated liver enzymes. These findings provide clues to the adverse effects of In humans. Although the potential biological significance between PFOA and lsmall and subclinical in the general US population, our data suggest that it wouto monitor the liver enzymes of people with low level exposure to PFOA, particwho are obese.”Gallo et al. (C8 Health Project) “A small but clear linear association between PSerum concentrations and ALT, a marker of hepatocelluar injury, was observed 

Authors’ Interpretation of Data

population-based sample of individuals with exposure to perfluoroalkyl acids. are consistent with previous findings and warrant further investigation, in partpotential health consequence of long-term exposure and potential accumulatiOlsen and Zobel (Occupational) “Measured hepatic clinical chemistry enzymes 

consistently associated with employees’ serum PFOA concentrations adjusted 

alcohol, and either BMI or serum triglycerides. A weakly positive association balkaline phosphatase, ALT and GGT with PFOA was observed among the Decatparticipants. Several epidemiologic research studies of the Decatur workforce reported associations with liver disease (malignant or non-malignant conditionvariety of data sources Including death certificates, episodes of care, and self-r
Is ALT associated with the glomerular filtration ra
Alanine Catabolism

Alanine is also important in intertissue nitrogen transport as part of the glucose-alanine cycle (see above). Alanine's catabolic pathway involves a simple aminotransferase reaction that directly produces pyruvate. Generally pyruvate produced by this pathway will result in the formation of oxaloacetate, although when the energy charge of a cell is low the pyruvate will be oxidized to CO2and H2O via the TCA cycle. This makes alanine a glucogenic amino acid.
Is Clearance a Common Explanation Across Many of these Epidemiologica
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion. The measurements we obtain may afford only a 

byproduct of the underlying pharmacokinetics, systems, biology, and pathogthese are not entirely new concepts, their importance may be clearer today before.”
Pharmacokinetic Variability and the Miracle of Modern Analytic

Matthew Longnecker (NIEHS) Epidemiology 2006;17:350-35

“The impressive capabilities of modern analytical chemistry are being appliefrequency in epidemiologic studies. With lower exposures being evaluated, 

concentrations being measured, it seems opportune to bear in mind that a gof variation in measured levels among subjects may be accounted for by diffmetabolism and excretion (clearance). The measurements we obtain may aglimpse of a byproduct of the underlying pharmacokinetics, systems, biologpathogenesis. While these are not entirely new concepts, their importance today than ever before.”

Epidemiologic associations are not equivalent to causation.

May expecphysiologicassociation“Reverse c
Ingredients to Detect Associations Based on Clearance 

Large Study Populations“Low” exposures and concentrations (PFOA/PFOSThe miracle of modern analytical chemistry
Ingredients to Detect Associations Based on Clearance 

Large Study Populations“Low” exposures and concentrations (PFOA/PFOSThe miracle of modern analytical chemistry

Physiology


Effect of Dietary Factors on Plasma PFO

Non‐HDL Cholesterol in ApoE*3.Leiden.CE

John Butenhoff, Ph.D., DABT

Geary Olsen, D.V.M, Ph.D.

Sue Chang, Ph.D.
BACKGROUND

 Cross

‐sectional studies have observed positive

associations of serum [PFOA] and [PFOS] with non

cholesterol (non

‐HDL

‐C).

Associations Associations weaken substantially substantially at serum PFO

PFOS concentrations >50 ng/mL.

Toxicological and mechanistic studies suggest lo

cholesterol as potential consequence of exposure 

an

d PFOS.

 Longitudinal Longitudinal occupational occupational study did not find as

as

of increasing serum [PFOA] and [PFOS] with non

‐H
HYPOTHESIS

 Serum lipid proteins may serve as a storage compartmPFOS and PFOA. Differences in dietary composition may lead to similar 

causal associations between non‐HDL‐C and PFOA/PFOS f

levels of exposure in diet.
STUDY DESIGN

 Four dietary dose groups (n=10 ♂ ApoE*3.Leiden.CETP 

Study Week

‐4 ‐3 ‐2 ‐10 123 4

No PFOA/PFOS added to diets ~30 ppb PFOA /~30 ppb PFO

added to diets

 [PFOA] in diets were 81 ± 4% of targeted concentration. [PFOS] in diets were 57 ± 27% of targeted concentration

Dietary Target Dietary [PFOA] d Mean M d Mean M d y Group [PFOA] and [PFOS], ng/g Measured [PFOA], ng/g Measured [PFOS], ng/g

1 36 28.4 17.8

2 34 27 4. 12 1.3 32 25.3 15.54 30 26.2 28.8
ApoE*3‐Leiden.CETP MICE

 Human‐like lipoprotein profiles and lipid levels ↓ clearance of apoB‐containing lipoproteins. Atherosclerosis characteristics of human lesion Response to treatment with drugs (i.e., statins, 

and niacin) at similar dosages to humans.
DIET COMPOSITION (kCal/g d

6534

l/g Diet

2

Kca

01

123

Dietary Group
CALORIC CONSUMPTION

181416

e

1012

Kcal/d/mous

468

AverageK

024

123

Dietary Group
EXPERIMENT

Study Week

‐4 ‐3 ‐2 ‐10 12

No PFOA/PFOS added to diets ~30 ppb PFOA 

added 

ementRandomization X Body Weight X X X 

Food Intake X X X X X 

MeasureFood Intake X X X X X 

Plasma Lipids X X X HDL X X X 

MPlasma [PFOA] X X X X 
Study Week

‐4 ‐3 ‐2 ‐10 123 4

No PFOA/PFOS added to diets ~30 ppb PFOA /~30 ppb PFO

added to diets

2 02.53.0

ol/L)

Group baseline lipoprotein profile

14161820

l/L)

Total Cholesterol 

1.01.52.0

Cholesterol (mmo

Dietary Group 1Dietary Group 2Dietary Group 3Dietary Group 4

68

1012

olesterol(mmol

0.00.5

0 5 10 15 20 25

Fraction

VLDL LDL HDL

0246

Dietary Group 1 Dietary Group 2 Diet

Ch

Fraction

 There were no differences in baseline cholesterol conce

Dietary Group 1 Dietary Group 2 Diet

and profiles prior to placement on modified cocoa butter/s
Study Week

‐4 ‐3 ‐2 ‐10 123 4

No PFOA/PFOS added to diets ~30 ppb PFOA /~30 ppb PFO

added to diets

14161820

/L)

Total Cholesterol No

2 02.53.0

/L) Dietary Group 1

VLDL

200250

Se

8

1012

14

olesterol(mmol/

1.01.52.0

Cholesterol (mmolDietary Group 1 Dietary Group 2 Dietary Group 3 Dietary Group 4

HDL

0

50

100150

200

Dietary Gro

PFOA Concentration (ng/mL)

0246 Cho

0.00.5

0 5 10 15 20 25

LDL HDL

Dietary Group 1 Dietary Group 2 Diet

Fraction

 Prior to adding PFOA/PFOS in diets, notable differencedietary groups in cholesterol profiles and concentrations b

apparent.
Study Week

‐4 ‐3 ‐2 ‐10 123 4

20

No PFOA/PFOS added to diets ~30 ppb PFOA /~30 ppb PFO

added to diets

14161820

ol/L)

Total Cholesterol 

2.02.53.0

mol/L) Dietray Group 1 + PFOA/PFOS Dietary Group 2 + PFOA/PFOS

VLDL

150200250

ion (ng/mL)

68

1012

Cholesterol(mm 1.0 1.5

Cholesterol (mmDietary Group 2 PFOA/PFOS

Dietary Group 3 + PFOA/PFOSDietary Group 4 + PFOA/PFOS

LDL

HDL

0

50

100

Dietary 

PFOA Concentrat

024

Dietary Group 1 Dietary Group 2 Dieta

C

0.00.5

0 5 10 15 20 25

Fraction

LDL

 Mean plasma [PFOA] ranged from 65 – 100 ng/mL. Plasma cholesterol cholesterol levels and lipoprotein lipoprotein profiles profiles did n

significantly compared to Week 0 levels.
Week 0

(no PFOA/PFOS in diet)

Week 

(PFOA/PFOS i

2 02.53.0

l/L) Dietary Group 1

VLDL

2.53.0

l/L) 

VLDL

(no PFOA/PFOS in diet) (PFOA/PFOS i

0 51.01.52.0

Cholesterol (mmolDietary Group 1

Dietary Group 2Dietary Group 3Dietary Group 4

LDL HDL

1.01.52.0

Cholesterol (mmol

LDL

0.00.5

0 5 10 15 20 25

Fraction

0.00.5

0 5 10 

Fraction

1214161820

ol/L)

Total Cholesterol Non‐HDL‐C HDL‐C

150200250

tration (ng/mL)

Serum PFOA Concentration (t = Week 0)

14161820

l/L)

Total Cholesterol Non‐HDL

150200250

tration (ng/mL)

468

10

12

Cholesterol (mmo

0

50

100

Dietary Group 1 Dietary Group 2 Dietary Group 3 Dietary Group 4

PFOA Concent

468

1012

Cholesterol (mmo 050 100 

PFOA Concent

024

Dietary Group 1 Dietary Group 2 Dietary Group 3 Dietary Group 4

024

Dietary Group 1 Dietary Group 2 Die
Study Week

‐4 ‐3 ‐2 ‐10 123 4

20

3 0

No PFOA/PFOS added to diets ~30 ppb PFOA /~30 ppb PFO

added to diets

12141618

mmol/)

Total Cholesterol 

2.02.53.0

mmol/L) Dietary Group 1 + PFOA/PFOS Di t G 2 + PFOA/PFOS

VLDL

152025

entration (ng/mL)

68

10

12

Cholesterol(m

1.01.5

Cholesterol(Dietary Group 2 + PFOA/PFOS

Dietary Group 3 + PFOA/PFOSDietary Group 3 + PFOA/PFOS

LDL

HDL

5

10

PFOA Conce

024

Dietary Group 1 Dietary Group 2 Dieta

0.00.5

0 5 10 15 20 25

Fraction

 Mean plasma PFOA continued to rise (128 – 195 ng/mL Plasma cholesterol cholesterol levels and lipoprotein lipoprotein profiles profiles did no

significantly compared to Week 0 and Week 2 levels.
Week 0

(no PFOA/PFOS in diet)

Week 

(PFOA/PFOS 

2 02.53.0

l/L) Dietary Group 1

VLDL

(no PFOA/PFOS in diet) (PFOA/PFOS 

2.53.0

l/L)

VLDL

0 51.01.52.0

Cholesterol (mmolDietary Group 1

Dietary Group 2Dietary Group 3Dietary Group 4

LDL HDL

1.01.52.0

Cholesterol (mmol

LDL

0.00.5

0 5 10 15 20 25

Fraction

0.00.5

0 5 10 

Fraction

LDL

1214161820

ol/L)

Total Cholesterol Non‐HDL‐C HDL‐C

150200250

ration (ng/mL)

Serum PFOA Concentration (t = Week 0)

1214161820

ol/)

Total Cholesterol Non‐HDL

tration (ng/mL)

468

10

12

Cholesterol (mmo

0

50

100

Dietary Group 1 Dietary Group 2 Dietary Group 3 Dietary Group 4

PFOA Concentr

468

10

12

Cholesterol (mm 

PFOA Concent

024

Dietary Group 1 Dietary Group 2 Dietary Group 3 Dietary Group 4

024

Dietary Group 1 Dietary Group 2 Die
16

1214

l, mmol/LWeek

0 Week

2 Week

4

8

10

Cholesterol

468

Non‐HDL‐C

024

Plasma N DG1 DG2 DG3 

0

01234

Ratio, Dietary kCal from Cocoa Butter:Dietary kCal from S

↑ kCal from Sucrose

↑ kC l a

l

from

Cocoa

Butter
250

200

ppb)

Week 1 Week 2 Week 4

150

], ng/mL(p

100

ma [PFOA] Plas 50

DG1 DG2 DG3 

0

01234

Ratio, Dietary kCal from Cocoa Butter:Dietary kCal from 

↑ kC l f C B

DG1 DG2 DG3 

↑ kCal from Sucrose ↑ kCal from Cocoa Butter
Week 2 Week 4

1416

DG1

1012

, mmol/L DG2

68

Non‐HDL‐C,

DG3

246

PlasmaNDG4

02

0 50 100 150 

Plasma PFOA Concentration, ng/mL
POSSIBLE MODE OF ACTION

 Fructose and glucose are known to stimulate lipogeneproduction of VLDL cholesterol; therefore, increasing the of cal i or es from sucrose may have result de the up‐regul 

a 

lipogenic pathways.

 In addition, with respect to plasma [PFOA], it is concePFOA, which resembles fatty acids, may compete with dieGI uptake as shown based on the bioavailability bioavailability chart.

 Both factors may have been influential in determining 

associ ti a ons f d oun in this st du y.
CONCLUSION

 Differences in isocalorically‐balanced dietary proportioand cocoa butter were shown to produce non‐causal, crospositive associations between plasma concentrations of noand PFOA in ApoE*3.Leiden.CETP 3.Leiden.CETP mice. Uncontrolled differences in dietary composition may be modifier modifier in cross‐sectional sectional epidemiological epidemiological associations associations in 

low serum concentrations of PFOA and, perhaps, other pe
THANK YOU
Six‐Month Capsule‐Dosing Study 

PFOS in Cynomolgus Monkey
Six‐Month Study of PFOS in

Cynomolgus Monkeys with Reco

• Daily intragastric capsule dosing

– Control, 0.03 mg/kg, 0.15 mg/kg, 0.75 mg/kg

• 5‐fold increments in dose

• Males and females females, 6/sex/group 6/sex/group except 0 03 . mg• Repeated clinical measures over 6‐months of do

– 5 to 6 time points for most measurements

• Hematology, serum clinical chemistry, thyroid hormones, 

• Baseline values for most measurements

– Allowed tracking of individual, within group, and be

group changes

• Complete histological evaluations

• Recovery Recovery sub‐groups with liver biopsy at 7 mont
Effects on Body Weight and Liver Weig

Relation to Dose and Serum [ 

PFOS

220 000240,000

Group Mean Serum [PFOS] in Cynomolgus Monkeys

after Six‐Months of Dosing

30

Mean Percent Change in Baseline Body We

Monkeys after Six Months of Dosing with 

100,000120,000140,000160,000180,000200,000220,000

PFOS], ng/mL (ppb) 05101520

25

Percent

0

20,00040,00060,00080,000

0 0.03 0.15 0.75

Serum[P

Dose Group, mg/kg/d

‐20‐15‐10

‐5

0 0.03 0.15 

Dose Group, mg/kg/d

p, g/ g/

3

3.5

Liver Weight to Body Weight Percent in Cyno

Monkeys after Six‐Months of Dosing with 

120140

Liver Weight in Cynomolgus Monkeys after Six

Months of Dosing with PFOS

p, g/ g/

1

1.5

2

2.5

Percent

406080

100

Liver Weight,g*

0

0.5

0 0.03 0.15 

Dose Group, mg/kg/d

0

20

0 0.03 0.15 0.75

Dose Group, mg/kg/d
Histopathology

• Histop gy athology

– Tissues examined

• All tissues from 0.75 mg/kg group by light microsco• Liver, thymus, and spinal cord from 0 03 . and 0 15 . m

by light microscopy

• All livers by transmission electron microscopy

– No treatment treatment‐related related findings findings at 0 03 . and 0 15 . – At 0.75 mg/kg

• LM: Centrilobular hypertrophy, vacuolation and mil• EM: ~50% incidence of l d ipi ‐d l rop et accumulation a

increased glycogen

– Resolved by 7 months in recovery
Liver Cell Electron Microscopy in M

Cynomolgus Cynomolgus Monkeys Monkeys of Six‐Month 

Control @ 6 mos. 0.75 mg/kg Male @ 6 mos. 0.75 mg/kg M

Recov
Serum‐Based Liver Toxicity Toxicity T

• No increases increases in serum activities activities of:

– Alanine aminotransaminase (ALT)– Aspartate aminotransaminase (AST)– γ‐Glutamyl transferase (GGT)– Alk li a ne ph ht osp a ase (ALP)– Sorbitol dehydrogenase (SDH)

• No indications of overt liver toxicity
Male Monkey Liver ALT and 

‐27 37 62 91 153 

Group

(mg/kg)

Study Day

0 32 39 34 35 43 

0.03 36 40 35 60 36 0.15 69 50 37 45 55 

ALT

(IU/L)

0.75 89 46 43 44 52 

0 56 45 36 39 31 

0 03 44 37 35 35 30 

( /)

AST 0.03 44 37 35 35 30 

0.15 72 48 40 42 40 0.75 68 43 34 36 39 

AST

(IU/L)

The only statistically significant changes observed were for decreased aeither within group from baseline or between treated groups and theimatched controls.These data argue against the presence of overt liver toxicity.
Female Monkey Liver ALT and 

27 37 62 91 153 

Group

(mg/kg)

Study Day

‐27 37 62 91 153 

0 140 59 53 54 77 

0.03 103 70 77 79 80 

(mg/kg)

ALT

/ 0.15 110 68 60 67 66 

0.75 123 46 42 45 48 

0 64 44 38 44 36 

(IU L)

0.03 54 49 45 43 38 0.15 74 36 37 37 36 0.75 72 36 31 34 40 

AST

(IU/L)

The only statistically significant changes observed were for decreased ac

within group from baseline or between treated groups and their time‐

controls.These data argue against the presence of overt liver toxicity.
Serum Lipids

• Decreased total cholesterol cholesterol (CHOL) and

decreased high‐density lipoprotein cho

(HDL‐C) in males and females females at 0 75 . m

dose.

• No baseline baseline values for HDL.
Male Monkey Serum Lipid

‐27 37 62 91 153 182

0 138 140 153 154 154 152

Study Day Comm

Statistically significant differ

Group

(mg/kg)

0.03 110 118 114 126 120 1100.15 151 146 144 150 149 1470.75 138 130 125 112 65 48

0 69 63 No baseline values. Differen

mg/kg group and controls b

baseline. Reductions in 0.75 

91 ‐ 182 are different than b

CHOL

(mg/dL)

0.03 46 420.15 55 480.75 19 13

0 2.2 2.4

mg/kg are in rough proport

matched CHOL (see ratio be

control at 0.75 mg/kg are lik

Values for 0.75 mg/kg grou

HDL‐C

(mg/dL)

0.03 2.6 2.60.15 2.8 3.20.75 3.4 3.7

0 70 69 62 68 60 45

different from control value

0, 0.03, and 0.15 mg/kg gro

specific HDL effect in 0.03 a

Reductions in TRIG from ba

CHOL/

HDL‐C

0.03 61 47 60 54 60 330.15 56 58 57 50 65 360.75 67 91 76 73 60 30

0.15 mg/kg and 0.75 mg/kg 

obvious effect of treatment

TRIG

(mg/dL)

= Statistically significantly different than within‐group baseline value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than time‐matched control value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than within‐group baseline AND time‐matched control values (Dunnett's t‐tes
Female Monkey Serum Lipi

‐27 37 62 91 153 182

0 149 147 155 166 163 160

Comm

Difference from control valu

Group Study Day

(mg/kg)

0.03 130 124 127 134 110 1220.15 144 133 137 140 130 1290.75 154 130 127 111 91 82

0 59 56

on Day 153 is not different t

Decreased CHOL in 0.75 mg

and 182 both compared to No baseline values. Differen

CHOL

(mg/dL)

0.03 47 420.15 41 360.75 23 21

0 2.8 2.9

in rough proportion to diffe

CHOL (see ratio below).No significant differences be

HDL‐C

(mg/dL)

0.03 2.4 3.00.15 3.3 3.90.75 4.0 4.4

0 73 71 86 67 80 56 No evidence for treatment‐r

and control. Similarities of r

and 0.75 mg/kg groups sug

effect on HDL.

CHOL/

HDL‐C

0.03 62 64 76 61 82 490.15 58 57 64 56 69 490.75 61 66 61 60 64 53

TRIG

(mg/dL)

= Statistically significantly different than within‐group baseline value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than time‐matched control value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than within‐group baseline AND time‐matched control values (Dunnett's t‐tes
Serum Thyroid Hormones

• Assay issues leading leading to reanalysis reanalysis by Ma

Clinic endocrinology unit.

• No cli i ll n ca y si ifi gn cant change in th i

yro

homeostasis based on reanalysis of sam

by Mayo Cli i n c.

• No histological evidence of thyroid chan

consistent with altered thyroid homeos
Male Monkey Thyroid Hormo

Group Study Day

‐27 37 62 91 182 184 184

0 0.00 0.34 0.57 0.00 0.43 0.37 0.55

0.03 0.00 0.35 0.42 0.00 0.34 0.56 0.56

0 15 0 05 0 33 0 98 0 03 0 74 0 70 1 38

Co

Problems with as

Mayo Clinic valu

reliable and show 

Study Day

TSH

( U/ L)

Group

(mg/kg)

0.15 0.05 0.33 0.98 0.03 0.74 0.70 1.380.75 0.00 0.21 0.22 0.00 0.93 0.93 1.43

0 1.01 1.40

0.03 1.21 1.530 15 0 94 1 47

differences from 

Negative bias in 

lab may have bee

dialysis‐RIA meth

(μU/mL)

FT4

( /dL) 0.15 0.94 1.47

0.75 1.12 1.50

0 3.49 4.06 2.27 3.89 4.38 3.25 3.24

0.03 4.46 5.11 3.91 5.30 4.72 3.85 3.680 15 4 63 4 00 3 08 4 47 3 99 2 71 3 00

demonstrated no 

No treatment‐re

evident. TT4

(μg/dL)(ng/dL)

0.15 4.63 4.00 3.08 4.47 3.99 2.71 3.000.75 4.88 3.61 2.59 4.61 5.34 3.30 3.77

0 4.21

0.03 5.130 15 4 33

Although FT3 gro

lower than contr

affected method

(μg/dL)

FT3

(pg/mL) 0.15 4.33

0.75 2.45

0 110 273 145 153 160 115 146

0.03 113 273 139 153 119 110 145

0 15 118 255 142 147 125 94 129

PFOS at 0.75 mg/

comparisons wit

controls, and con

equivocal. (pg/mL)

TT3

(ng/dL)

= Statistically significantly different than within‐group baseline value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than time‐matched control value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than within‐group baseline AND time‐matched control values (Dunnett's t‐tes

0.15 118 255 142 147 125 94 1290.75 133 239 121 118 66 43 76

Physiological con

(ng/dL)
Female Female Monkey Monkey Thyroid Thyroid Horm

Horm

Group Study Day

‐27 37 62 91 182.00 184.00 184.00

0 0.21 0.73 1.19 0.00 0.73 0.53 1.02

0.03 0.00 0.24 0.46 0.00 0.68 0.43 2.01

0 15 0 34 0 61 0 82 0 02 1 27 0 47 1 33

Co

Problems with co

changes in TSH b

Group

(mg/kg)

TSH

( U/ L)

Study Day

0.15 0.34 0.61 0.82 0.02 1.27 0.47 1.330.75 0.01 0.59 0.74 0.00 0.84 1.03 1.86

0 1.06 1.53

0.03 1.01 1.700 15 0 90 1 35

No changes in FT

(μU/mL)

FT4

( /dL) 0.15 0.90 1.35

0.75 1.08 1.57

0 6.46 5.82 3.53 4.77 5.66 4.14 3.78

0.03 7.06 5.06 4.07 6.00 4.33 3.48 2.80

0 15 7 13 3 68 2 77 3 77 3 91 2 88 3 23

No changes in TT

(ng/dL)

TT4

(μg/dL) 0.15 7.13 3.68 2.77 3.77 3.91 2.88 3.23

0.75 7.10 4.23 3.08 4.92 5.61 3.43 3.80

0 4.05

0.03 3.59

0 15 3 27

Although FT3 gro

lower than contr

affected method

(μg/dL)

FT3

(pg/mL) 0.15 3.27

0.75 2.82

0 119 230 115 143 135 106 148

0.03 171 244 136 147 120 92 139

0 15 138 214 114 110 97 80 116

Mayo Clinic data 

significant differe

controls at term. equivocal.

TT3

(ng/dL)

(pg/mL)

= Statistically significantly different than within‐group baseline value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than time‐matched control value (Dunnett's t‐test, p < 0.05)

= Statistically significantly different than within‐group baseline AND time‐matched control values (Dunnett's t‐tes

0.15 138 214 114 110 97 80 1160.75 155 196 96 88 85 58 99

0.75 mg/kg is equ

(ng/dL)
Steroid Hormones

• Steroid Steroid hormones hormones (cortisol cortisol, estrogen estrogen, e

e

estrone, testosterone)

– Only observed observed decreased decreased estradiol estradiol and on

0.75 mg/kg males
Other Measures

• No toxicologically toxicologically meaningful meaningful changes changes i

– Other clinical chemistry measures

• SBA, CPK, BILI, BUN, GLU, CREAT, PROT, ALB, G

CL, K, NA,

– Hematological parameters– Urinalyses– PCNA labe g lin index ( e, liv r pa c eas, pancreas, tes e

t

– Hepatic palmitoyl CoA oxidase
Thank You

Questions and Comments