Grantee Research Project Results

Final Report: Dioxins, Male Pubertal Development and Testis Function

EPA Grant Number: R829437
Title: Dioxins, Male Pubertal Development and Testis Function
Investigators: Hauser, Russ , Korrick, Susan A. , Williams, Paige L.
Institution: Harvard University
EPA Project Officer: Chung, Serena
Project Period: December 1, 2001 through November 30, 2005 (Extended to February 3, 2008)
Project Amount: $2,252,427
RFA: Endocrine Disruptors: Epidemiologic Approaches (2001) RFA Text | Recipients Lists
Research Category: Endocrine Disruptors , Environmental Justice , Human Health , Safer Chemicals

Objective:

Specific Aim #1: To investigate whether alterations in physical growth and sexual maturation in boys are associated with in utero levels of dioxins and/or serum dioxin concentrations measured in adolescence. In utero dioxin levels will be estimated using the child’s current serum dioxin level, as well as historical reproductive (e.g. maternal breast feeding) and residential information.

Note: Described in detail of summary of results.

Specific Aim #2: To retrospectively investigate whether in utero levels of dioxins are associated with reproductive tract abnormalities, specifically hypospadias, cryptorchidism and testicular abnormalities (manifested by small testicular volume at sexual maturity).

Note: The retrospective estimation of in utero levels of dioxins is underway. A doctoral student (Olivier Humblet) is working on this aim as a main component of his thesis, with completion anticipated in the Fall of 2009. As part of this aim, in the Spring of 2009 we resampled a small subset of women from whom we now have blood samples for dioxins that are 9 years apart. This will allow us to use this data to validate our pre-natal exposure estimation models.

Specific Aim #3: To investigate whether serum dioxins measured in adolescence are associated with disturbances in testis function, specifically endocrine function and spermatogenesis.

Note: Due to the delays in funding and revision of study design (see below for details) we were unable to investigate associations of dioxins with semen quality. To assess onset of puberty in our study, by design we recruited children at age 8 to 9 years old and therefore they are currently only 14 to 15 years of age, not old enough for semen collection. We have NIEHS funding that will allow us to continue to follow these children and ultimately we will apply for NIEHS renewal funding that will allow us to assess semen quality when they reach 18 years of age

Summary/Accomplishments (Outputs/Outcomes):

The specific aims outlined in the grant proposal were modified due to difficulties in initiating a large international study in Russia. During the first year of the project, we purchased the necessary equipment and supplies, trained personnel and developed the databases that will be used through-out the study. We also experienced (and reported to EPA) very long delays in shipping the supplies to Russia, a direct result of the extensive paperwork needed for Russian customs, as well as U.S. shipping permits. It took almost a year to get the necessary approvals from the Russian government.

As originally proposed in the grant application, we were planning on following the children enrolled in our earlier pilot study (1999-2000). However, because of the delay in receiving funding (delayed from July 2001 until February 2002, due to the international scope of the project in Russia) and the delay in getting permission to send supplies to Russia, we had no choice but to assemble a new cohort of 8 and 9-year old boys rather than following the boys from the pilot study. The boys in the pilot study, as a result of the delays in the start of funding and Russian governmental delays, were too old to follow for growth and onset of sexual maturation. However, our pilot data was useful in guiding us in the optimal design of the prospective cohort study of 500 nine-year old boys. Recruitment of boys in the prospective cohort study began in May 2003.

We realized this was a change in the proposed design, but the delays were out of our control (delayed EPA funding start date – approximately 7 months) as well as unanticipated delays in procuring Russian customs approval to ship supplies (partly a result of the new post September 11^th concerns with air travel and shipping unknown supplies-approximately 8 months delay).

However, despite the delays and revision of the aims, we are still able to investigate our primary aim of the association of dioxins with alterations in physical growth and sexual maturation. The endpoints include growth velocity, onset and tempo of sexual maturation, as well attainment of sexual maturity. We believe that this change in the design of subject recruitment and follow-up was optimal and an improvement over the proposed study plan. This was discussed with Dr. David Reese in the fall and winter of 2002 and he approved our proposed changes.

Conclusions:

Between May 2003-2005, we recruited 516 boys and their families to participate in our prospective cohort study on growth and pubertal development. It is important to keep in mind that when conducting a large prospective cohort study, such as ours, it takes many years of follow-up and subsequent data analysis to generate results and publications. Although we are in year 6 of the study, we anticipate many publications and interesting results in the coming years.

1). Study participant follow-up and collection and analysis of data

Subject follow-up remains excellent and our plans include following the children until adulthood (age 18 years, so that we can assess attainment of sexual maturation and semen quality). Each year we complete annual follow-up physical examinations, urine collection and questionnaires (health, lifestyle, diet). Every other year, we collect blood from the boys. The end of year 3 follow-up was completed for all boys with an overall retention rate of 87%.

Dietary Analysis: We completed work with the Russian Institute of Nutrition to analyze the dietary intake information on the food frequency questionnaire. Dr. Jane Burns, a research associate on the project, has taken the lead in the dietary analysis and presented a poster at the ISEE/ISEA 2006 meeting in Paris, France.

Biological Samples: The analysis of dioxins, furans and PCBs by the CDC for all the baseline samples (i.e., boys and their mothers) has been completed. In addition, the baseline samples were analyzed for the organochlorine pesticides hexachlorabenze (HCB), beta-hexachlorocyclohexane (bHCCH), and dichlorodiphenyldichloroethylene (p,p'-DDE). In addition, we analyzed all baseline samples for blood lead.

In table 1, we present the results for serum PCDD/PCDF/C-PCBs (pg/g lipid) concentrations and 2005 WHO TEQs among 8-9 year old boys in the Russian Children’s Study (N=482)

Table 1.

			Concentrations ^b			2005 WHO TEQs (pg TEQ/g lipid) Percentiles
	Median LOD^a	% of samples below LOD	25th	Percentiles Median	75th	25th	Median	75th
PCDDs (pg/g lipid)
2,3,7,8-TCDD	0.60	26%	1.34	2.75	3.90	1.34	2.75	3.90
1,2,3,7,8-PeCDD	0.70	28%	1.41	4.10	7.00	1.41	4.10	7.00
1,2,3,4,7,8-HxCDD	1.10	63%	0.71	2.00	3.90	0.07	0.20	0.39
1,2,3,6,7,8-HxCDD	1.10	12%	5.40	8.70	16.6	0.54	0.87	1.66
1,2,3,7,8,9-HxCDD	1.10	52%	0.85	2.61	4.60	0.09	0.26	0.46
1,2,3,4,6,7,8-HpCDD	1.20	<1%	8.20	12.2	19.5	0.08	0.12	0.20
OCDD	16.1	26%	69.0	96.1	134	0.02	0.03	0.04
PCDFs (pg/g lipid)
2,3,7,8-TCDF	0.70	91%	0.42	0.50	1.63	0.04	0.05	0.16
1,2,3,7,8-PeCDF	0.70	83%	0.42	0.57	1.91	0.01	0.02	0.06
2,3,4,7,8-PeCDF	0.60	3%	6.20	9.0	14.6	1.86	2.70	4.38
1,2,3,4,7,8-HxCDF	0.70	2%	4.10	6.65	12.5	0.41	0.67	1.25
1,2,3,6,7,8-HxCDF	0.70	11%	2.90	4.20	6.70	0.29	0.42	0.67
1,2,3,7,8,9-HxCDF	0.80	99%	0.42	0.57	1.41	0.04	0.06	0.14
2,3,4,6,7,8-HxCDF	0.70	88%	0.42	0.57	1.84	0.04	0.06	0.18
1,2,3,4,6,7,8-HpCDF	0.80	7%	5.47	7.50	11.3	0.06	0.08	0.11
1,2,3,4,7,8,9-HpCDF	0.80	85%	0.50	0.64	2.26	0.01	0.01	0.02
OCDF	0.90	26%	1.80	2.90	5.00	0.001	0.001	0.002
Co-planar PCBs (pg/g lipid)
3,3',4,4'-TCB 77	1.40	0%	48.9	88.1	134	0.005	0.009	0.01
3,4,4′,5-TCB 81	1.50	3%	5.90	8.45	12.6	0.002	0.003	0.004
3,3′,4,4′,5-PeCB 126	1.50	<1%	40.3	58.0	84.4	4.03	5.80	8.44
3,3′,4,4′,5,5′-HxCB 1	1.30	2%	11.1	16.9	28.9	0.33	0.51	0.87
Mono-ortho PCBs (ng/g lipid)
2,3,3′,4,4′-PeCB (105)	0.30	<1%	4.90	7.40	11.1	0.15	0.22	0.33
2,3′,4,4′,5-PeCB (118)	0.30	<1%	22.4	33.1	48.7	0.67	0.99	1.46
2,3,3′4,4′,5-HxCB (156)	0.40	1%	3.60	5.80	10.7	0.11	0.17	0.32
2,3,3′,4,4′,5′-HxCB (157)	0.40	14%	1.00	1.80	3.30	0.03	0.05	0.10
2,3′,4,4′,5,5′-HxCB (167)	0.50	11%	1.30	2.10	3.45	0.04	0.06	0.10
2,3,3′,4,4′,5,5′-HpCB (189)	0.60	73%	0.21	0.57	0.78	0.006	0.02	0.02
Total PCDDs (pg/g lipid)			93.3	136	189	4.5	8.2	13.5
Total PCDFs (pg/g lipid)			26.7	38.9	57.3	3.0	4.2	6.9
Total co-planar PCBs (pg/g lipid)			126	181	249	4.5	6.4	9.4
Total PCDD/F/Co-planar PCBs (pg/g lipid)			278	362	499	13.3	19.6	30.5
Total Mono-ortho PCBs (ng/g lipid)			35.3	51.7	78.2	1.1	1.6	2.4
Total PCBs (ng/g lipid)			164	249	393
Total TEQs						14.4	21.1	33.2

^aMedian LOD (limit of detection): median of sample specific LOD for each congener;
^bSamples below the LOD were assigned a value = LOD/√2;
Congeners are identified according to the International Union for Pure and Applied Chemistry (IUPAC) nomenclature.

1. Manuscript, ‘Association of blood lead levels with onset of puberty in Russian boys’. Russ Hauser, Oleg Sergeyev, Susan Korrick, Mary M. Lee, Boris Revich, Elena Gitin, Jane Burns, Paige Williams was published in Environmental Health Perspectives 2008 Jul;116(7):976-80.

Background: Epidemiological studies suggest a temporal trend of earlier onset and longer duration of puberty, raising concerns regarding the potential impact of environmental factors on pubertal development. Lead exposure has been associated with delayed pubertal onset in girls; however, epidemiologic data in boys is limited.

Methods: We used multivariable logistic regression models to explore the cross-sectional association of blood lead levels with growth and pubertal onset based on physician-assessed testicular volume and pubertal staging in 489 eight- to nine-year old boys from Chapaevsk, Russia. Multivariable linear regression models were used to assess associations of blood lead levels with somatic growth at the study entry visit.

Results: The median (25^th, 75^th percentile) blood lead level was 3 µg/dL (2 µg/dL, 5 µg/dL). Height, weight, BMI, birth weight and gestational age were predictive of the onset of puberty as assessed either by testicular volume (greater than 3 ml), Genitalia Stage (G2), or both. Blood lead level was inversely associated with height (P< 0.001) and weight (P=0.06) after adjustment for birth weight, gestational age, and age at examination. In multivariable adjusted analyses, boys with blood lead levels of 5 µg/dL or higher had a 43% reduced odds of having entered G2 as compared to those with lower levels (OR=0.57; 95% CI: 0.34-0.95, p=0.03).

Conclusions: Relatively low environmental blood lead levels were associated with decreased growth and differences in pubertal onset in peri-adolescent Russian boys. Future analyses of this prospective cohort will address pubertal onset and progression in relation to lead and other environmental chemicals.

Table. Association of lead (natural log transformation) with measures of physical growth and birth characteristics among 8- to 9-year-old Russian boys (n=489) based on univariate and multiple linear regression models.

	Unadjusted Regression Coefficient			Adjusted Regression Coefficient**
	Estimate	95% CI	P-value	Estimate	95% CI	p-value

Height (cm)	-1.043	(-1.95,-0.13)	0.02	-1.439	(-2.25,-0.63)	<0.001
Weight (kg)	-0.764	(-1.57,0.04)	0.06	-0.761	(-1.54,0.02)	0.067
Body Mass Index	-0.206	(-0.54,0.13)	0.22	-0.107	(-0.44,0.23)	0.53
Penile Length (cm)	-0.004	(-0.11,0.10)	0.94	0.023	(-0.09,0.13)	0.68
Birth Weight (kg)	-0.094	(-0.17,-0.02)	0.02	-0.084	(-0.15,-0.02)	0.01
Gestational Age (wks)	-0.038	(-0.29,0.22)	0.77	0.118	(-0.09,0.33)	0.27

** - linear regression models adjusted for birthweight, gestational age, and age at exam (except for models for birthweight and gestational age, which are adjusted for all other listed effects)

Table. Odd ratios for the association of lead (natural log transformation and high lead) with puberty onset among 8- to 9-year-old Russian boys (n=489) based on logistic regression models.

Unadjusted regression coefficient

Adjusted regression coefficient**

95% CI

p-value

95% CI

p-value

Models for effect of Lead (natural log transformation) with puberty onset

Testicular Volume

All 6 levels*

0.80

(0.61-1.06)

0.12

0.90

(0.67-1.20)

0.47

Puberty Onset (volume > 3ml)

1.01

(0.67-1.53)

0.96

1.08

(0.69-1.70)

0.74

Tanner Staging

Tanner Stage ≥ G2

0.82

(0.60-1.12)

0.20

0.75

(0.53-1.06)

0.10

Tanner Stage ≥ P2

1.37

(0.81-2.33)

0.24

1.08

(0.60-1.93)

0.81

Models for effect of High Lead (5 μg/dL or higher) with puberty onset

Testicular Volume

All 6 levels*

0.57

(0.39-0.83)

0.004

0.72

(0.48-1.07)

0.11

Puberty Onset (volume > 3ml)

0.77

(0.42-1.40)

0.39

0.83

Journal Articles on this Report : 9 Displayed | Download in RIS Format

Publications Views
Other project views:	All 11 publications	9 publications in selected types	All 9 journal articles

Publications
Type	Citation	Project	Document Sources
Journal Article	Burns JS, Williams PL, Sergeyev O, Korrick S, Lee MM, Revich B, Altshul L, Turner WE, Patterson DG, Saharov I, Hauser R. Predictors of serum dioxins and PCBs in a Russian cohort of boys. Organohalogen Compounds 2008;70:1490-1493.	R829437 (Final)	not available
Journal Article	Burns JS, Williams PL, Sergeyev O, Korrick S, Lee MM, Revich B, Altshul L, Patterson Jr. DG, Turner WE, Needham LL, Shararov I, Hauser R. Predictors of serum dioxins and PCBs among peripubertal Russian boys. Environmental Health Perspectives 2009;117(10):1593-1599.	R829437 (Final)	Full-text from PubMed Abstract from PubMed Associated PubMed link Full-text: EHP
Journal Article	Hauser R, Williams P, Altshul L, Korrick S, Peeples L, Patterson Jr. DG, Turner WE, Lee MM, Revich B, Zeilert V, Sergeyev O. Characterization and predictors of serum dioxin levels among adolescent boys in Chapaevsk, Russia. Organohalogen Compounds 2004;66:3245-3251.	R829437 (2004) R829437 (2006) R829437 (2007) R829437 (Final)	not available
Journal Article	Hauser R, Williams P, Altshul L, Korrick S, Peeples L, Patterson DG Jr, Turner WE, Lee MM, Revich B, Sergeyev O. Predictors of serum dioxin levels among adolescent boys in Chapaevsk, Russia: a cross-sectional pilot study. Environmental Health: A Global Access Science Source 2005;4(1):8.	R829437 (2004) R829437 (2006) R829437 (2007) R829437 (Final)	Full-text from PubMed Abstract from PubMed Associated PubMed link Full-text: Environmental Health Exit Other: Environmental Health PDF Exit
Journal Article	Hauser R, Sergeyev O, Korrick S, Lee MM, Revich B, Gitin E, Burns JS, Williams PL. Association of blood lead levels with onset of puberty in Russian boys. Environmental Health Perspectives 2008;116(7):976-980.	R829437 (2007) R829437 (Final)	Full-text from PubMed Abstract from PubMed Associated PubMed link Full-text: EHP Exit Abstract: EHP Exit Other: EHP PDF Exit
Journal Article	Lee MM, Sergeyev O, Williams P, Korrick S, Zeilert V, Revich B, Hauser R. Physical growth and sexual maturation of boys in Chapaevsk, Russia. Journal of Pediatric Endocrinology & Metabolism 2003;16(2):169-178.	R829437 (2003) R829437 (2006) R829437 (2007) R829437 (Final)	Abstract from PubMed Abstract: Reference Global Exit
Journal Article	Milnes MR, Bryan TA, Medina JG, Gunderson MP, Guillette Jr. LJ. Developmental alterations as a result of in ovo exposure to the pesticide metabolite p,p '-DDE in Alligator mississippiensis. General and Comparative Endocrinology 2005;144(3):257-263.	R829437 (Final) R824760 (Final)	Abstract from PubMed Full-text: Science Direct - Full Text HTML Exit
Journal Article	Sergeyev O, Revich B, Williams P, Korrick S, Zeilert V, Lee MM, Ushakova T, Saharov I, Altshul L, Hauser R. A case-cohort study of cryptorchidism, hypospadias and delayed sexual maturation in a dioxin contaminated region: Chapaevsk, Russia. Organohalogen Compounds 2002;59:385-388.	R829437 (2004) R829437 (2006) R829437 (2007) R829437 (Final)	not available
Journal Article	Williams P, Sergeyev O, Lee M, Korrick S, Burns J, Humblet O, DelPrato J, Revich B, Haer R. Blood Lead Levels and Delayed Onset of Puberty in a Longitudinal Study of Russian Boys. PEDIATRICS 2010;125(5):E1088-E1096.	R829437 (Final)	Full-text from PubMed Abstract from PubMed Full-text: Pediatrics - Full Text HTML Exit Other: Pediatrics - Full Text PDF Exit

Supplemental Keywords:

Children, Dioxins, Health, Human, Lead, Puberty,, Health, RFA, Scientific Discipline, Toxics, Health Risk Assessment, pesticides, Risk Assessments, Biology, Endocrine Disruptors - Human Health, Molecular Biology/Genetics, Children's Health, endocrine disruptors, Biochemistry, Endocrine Disruptors - Environmental Exposure & Risk, cell-cell interactions, male sexual development, hypospadias, EDCs, physiology, embryonic development, dioxin, human growth and development, developmental biology, testis development, human exposure, chemical interference, postnatal development, male fertility, endocrine disrupting chemicals, paracine growth factors, fetal development, sexual development, sertoli cells, human health risk, gonad morphology, neurotropin growth, puberty

Progress and Final Reports:

Original Abstract

The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.