United States
Environmental Protection
Jf lkAgency

EPA/600/R-23/375
January 2024

www.epa.gov/isa

Integrated Science
Assessment for Lead

Appendix 8: Reproductive and
Developmental Effects

January 2024

Center for Public Health and Environmental Assessment

Office of Research and Development
U.S. Environmental Protection Agency


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DISCLAIMER

This document has been reviewed in accordance with the U.S. Environmental Protection Agency
policy and approved for publication. Mention of trade names or commercial products does not constitute
endorsement or recommendation for use.

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DOCUMENT GUIDE

This Document Guide is intended to orient readers to the organization of the Lead (Pb) Integrated
Science Assessment (ISA) in its entirety and to the sub-section of the ISA at hand (indicated in bold). The
ISA consists of the Front Matter (list of authors, contributors, reviewers, and acronyms), Executive
Summary, Integrated Synthesis, and 12 appendices, which can all be found at https://assessments.epa.gov/

i sa/ctoeume nt/&de i d=3 5 953 6.

Front Matter
Executive Summary
Integrated Synthesis

Appendix 1. Lead Source to Concentration

Appendix 2. Exposure, Toxicokinetics, and Biomarkers

Appendix 3. Nervous System Effects

Appendix 4. Cardiovascular Effects

Appendix 5. Renal Effects

Appendix 6. Immune System Effects

Appendix 7. Hematological Effects

Appendix 8. Reproductive and Developmental Effects

Appendix 9. Effects on Other Organ Systems and Mortality

Appendix 10. Cancer

Appendix 11. Effects of Lead in Terrestrial and Aquatic Ecosystems
Appendix 12. Process for Developing the Pb Integrated Science Assessment

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CONTENTS

DOCUMENT GUIDE 	8-iii

LIST OF TABLES	8-v

LIST OF FIGURES	8-vi

ACRONYMS AND ABBREVIATIONS	8-vii

APPENDIX 8 REPRODUCTIVE AND DEVELOPMENTAL EFFECTS	8-1

8.1	Introduction and Summary of the 2013 Integrated Science Assessment	8-1

8.1.1	Effects on Pregnancy and Birth Outcomes	8-2

8.1.2	Effects on Development	8-2

8.1.3	Effects on Female Reproductive Function	8-3

8.1.4	Effects on Male Reproductive Function	8-3

8.2	Scope	8-4

8.3	Effects on Pregnancy and Birth Outcomes	8-5

8.3.1	Maternal Health During Pregnancy	8-6

8.3.2	Prenatal Growth	8-10

8.3.3	Preterm Birth	8-17

8.3.4	Birth Defects	8-20

8.3.5	Spontaneous Abortion and Pregnancy Loss and Fetal and Infant Mortality	8-23

8.3.6	Placental Function	8-25

8.3.7	Other Pregnancy and Birth Outcomes	8-27

8.4	Effects on Development	8-29

8.4.1	Effects on Postnatal Growth 	8-29

8.4.2	Effects on Puberty among Females	8-34

8.4.3	Effects on Puberty among Males	8-37

8.4.4	Other Developmental Effects	8-40

8.5	Effects on Female Reproductive Function	8-41

8.5.1	Effects on Hormone Levels and Menstrual/Estrous Cycle	8-41

8.5.2	Effects on Female Fertility	8-45

8.5.3	Effects on Morphology and Histology of Female Sex Organs (Ovaries, Uterus,

Fallopian Tubes/Oviducts, Cervix, Vagina, and Mammary Glands)	8-47

8.6	Effects on Male Reproductive Function	8-48

8.6.1	Effects on Sperm/Semen Production, Quality, and Function	8-48

8.6.2	Effects on Hormone Levels in Males	8-51

8.6.3	Effects on Male Fertility	8-54

8.6.4	Effects on Morphology and Histology of Male Sex Organs	8-55

8.7	Biological Plausibility	8-57

8.7.1	Pubertal Onset	8-58

8.7.2	Male Reproduction Function	8-59

8.8	Summary and Causality Determination	8-61

8.8.1	Summary of Effects on Pregnancy and Birth Outcomes	8-61

8.8.2	Summary of Effects on Development	8-64

8.8.3	Summary of Effects on Female Reproductive Function	8-67

8.8.4	Summary of Effects on Male Reproductive Function	8-70

8.9	Evidence Inventories - Data Tables to Summarize Study Details	8-79

8.10	References	8-243

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LIST OF TABLES

Table 8-1	Summary of evidence contributing to causality determinations for Pb exposure and

reproductive and developmental effects	8-74

Table 8-2	Epidemiologic studies of exposure to Pb and maternal health outcomes	8-79

Table 8-3	Animal toxicological studies of Pb exposure and pregnancy and birth outcomes	8-94

Table 8-4	Epidemiologic studies of Pb exposure and prenatal growth	8-99

Table 8-5	Epidemiologic studies of Pb exposure and preterm birth	8-134

Table 8-6	Epidemiologic studies of Pb exposure and birth defects 	8-146

Table 8-7	Epidemiologic studies of Pb exposure and fetal and infant mortality and spontaneous

abortion and pregnancy loss	8-153

Table 8-8	Epidemiologic studies of Pb exposure and placental function	8-157

Table 8-9	Epidemiologic studies of Pb exposure and other pregnancy and other birth outcomes	8-159

Table 8-10 Epidemiologic studies of Pb exposure and postnatal growth	8-165

Table 8-11 Animal toxicological studies of Pb exposure and development	8-184

Table 8-12 Epidemiologic studies of exposure to Pb and puberty in females and puberty in males	8-191

Table 8-13 Epidemiologic studies of exposure to Pb and other developmental effects	8-204

Table 8-14 Epidemiologic studies of exposure to Pb and female reproductive effects	8-210

Table 8-15 Animal toxicological studies of Pb exposure and female reproductive effects	8-220

Table 8-16 Epidemiologic studies on exposure to Pb and male reproductive effects	8-223

Table 8-17 Animal toxicological studies of exposure to Pb and male reproductive effects	8-240

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LIST OF FIGURES

Figure 8-1	Potential biological pathways for reproductive and developmental effects following

exposure to Pb. 	8-58

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ACRONYMS AND ABBREVIATIONS

2PN	oocytes with two pronuclei

AAS	atomic absorption spectrometry

AD	abdominal diameter

AGD	anogenital distance

AGDap	anopenile distance

AGDas	anoscrotal distance

ALAD	S-aminolevulinic acid dehydratase

ALSPAC	Avon Longitudinal Study of Parents

and Children

AMH	anti-Mullerian hormone

AQCD	Air Quality Criteria Document

ART	assisted reproductive technology

As	arsenic

BKMR	Bayesian kernel machine regression

BL	birth length

BLL	blood lead level

BMI	body mass index

BMIZ	BMI-for-age Z-score

BT20+	Birth to Twenty Plus

BW	birth weight

BWGA	birth weight-for-gestational age

BWZ	birth weight Z-score

C-ABCS	China-Anhui Birth Cohort Study

CANDLE	Conditions Affecting Neurocognitive
Development and Learning in Early
Childhood

CC	chest circumference

CCG	Charlotte-Concord-Gastonia

Cd	cadmium

CD	cephalic diameter

CHD	congenital heart disease

CHECK	Children's Health and Environmental
Chemicals in Korea

CHL	crown-heel length

CI	confidence interval

CMS	Charlotte Motor Speedway

Cr	chromium

d	day(s)

DBP	diastolic blood pressure

E2	estradiol

E3G	estrone-3-glucuronide

EAAS	electrothermal atomic absorption
spectrometry

ELEMENT	Early Life Exposure in Mexico to
Environmental Toxicants

ELISA	enzyme-linked immunosorbent assay

EMASAR	Study on the Environment and

Reproductive Health

e-REACH	e-waste Recycling Exposure and

Community Health

ETS	environmental tobacco smoke

fE2	free estradiol

FLEHS	Flemish Environment and Health Study

FSH	follicle stimulating hormone

FT	free testosterone

FT3	free triiodothyronine

FT4	free thyroxine

GA	gestational age

GD	gestational day

GDM	gestational diabetes mellitus

GEE	generalized estimating equation

GFAAS	graphite furnace atomic absorption
spectrometry

GnRH	gonadotropin-releasing hormone

GSI	Global Severity Index

HAZ	height-for-age Z-score

HC	head circumference

HCAZ	head circumference for age Z-score

hCG	human chorionic gonadotropin

HFIAS	Household Food Insecurity Access

Scale

Hg	mercury

HOME	Health Outcomes and Measures of the

Environment

HR	hazard ratio

HR-ICP-MS high resolution inductively coupled

plasma mass spectrometry
HTZ	height Z-score

hr	hour(s)

ICP-AES	inductively coupled plasma atomic

emission spectroscopy
ICP-MS	inductively coupled plasma mass

spectrometry

ICP-QQQ	inductively coupled plasma triple quad

IgE	immunoglobulin E

IGF-1	insulin-like growth factor 1

IGT	impaired glucose tolerance

IL-33	interleukin-33

INMA	Instituto de Nanociencia y Materiales

de Aragon

IQR	interquartile range

ISA	Integrated Science Assessment

IUGR	intrauterine growth restriction

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IVF	in vitro fertilization

JECS	Japan Environment and Children's

Study

K6	Kessler Psychological Distress Scale

KNHANES	Korea National Health and Nutrition

Examination Survey

K-XRF	K-shell X-ray fluorescence

LA-ICP-MS laser ablation-inductively coupled

plasma-mass spectrometry
LBW	low birth weight

LESPW	Life Event Scale for Pregnant Women

LGA	large for gestational age

LH	luteinizing hormone

LIFE	Longitudinal Investigation of Fertility

and the Environment
LMP	last menstrual period or last missed

period

In	natural log

LOD	limit of detection

MAL-ED	Interactions of Malnutrition and Enteric

Infections: Consequences for Child
Health and Development
Mil	metaphase II

min	minute(s)

MIREC	Maternal-Infant Research on

Environmental Chemicals
miRNA	micro RNA

MMP	matrix metalloproteinase

Mn	manganese

mo	month(s)

MOCEH	Mothers'and Children's

Environmental Health

MSA	Metropolitan Statistical Area

mtDNA	mitochondrial DNA

mtDNAcn	mitochondrial DNA copy number

NASCAR	National Association for Stock Car

Auto Racing
NHANES	National Health and Nutrition

Examination Survey

NICE	Nutritional impact on Immunological

maturation during Childhood in relation
to the Environment

NR	not reported

NS	non-stress

NTD	neural tube defect

OFC	orofacial cleft

OGTT	oral glucose tolerance test

OR	odds ratio

Pb	lead

PECOS	Population, Exposure, Comparison,
Outcome, and Study Design

PI	Ponderal Index

PIR	poverty-income ratio

PM2.5	fine particulate matter

PND	postnatal day

PROGRESS	Programming Research in Obesity,
Growth, Environment and Social
Stressors

PROM	premature rupture of membranes

PROTECT	Puerto Rico Test site for Exploring
Contamination Threats

QL	lower quartile

QUS	quantitative ultrasound

ROS	reactive oxygen species

RR	relative risk

rTL	relative telomere length

SA	semen analysis

SBP	systolic blood pressure

SCL-90-R	Symptom-Checklist-90-Revised

SD	standard deviation

Se	selenium

SE	standard error

SES	socioeconomic status

SGA	small for gestational age

SHBG	sex hormone binding globulin

SNP	single nucleotide polymorphism

SPECT	Survey on the Prevalence in East China
for Metabolic Diseases and Risk
Factors

T	testosterone

T#	tertitle #

TL	telomere length

TPOAb	thyroid peroxidase antibody

TRI	Toxics Release Inventory

TSH	thyroid-stimulating hormone

TSLP	thymic stromal lymphopoietin

tT	total testosterone

tT3	total triiodothyronine

tT4	total thyroxine

TV	testicular volume

UCB	umbilical cord blood

WAZ	weight for age Z-score

WC	waist circumference

wk	week(s)

WHEALS	Wayne County Health, Environment,
Allergy and Asthma Longitudinal
Study

WHO	World Health Organization

yr	year(s)

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APPENDIX 8 REPRODUCTIVE AND

DEVELOPMENTAL EFFECTS

Summary of Causality Determinations for Pb Exposure and
Reproductive and Developmental Effects

This appendix characterizes the scientific evidence that supports causality
determinations for Pb exposure and reproductive and developmental effects. The types
of studies evaluated within this appendix are consistent with the overall scope of the
ISA as detailed in the Process Appendix (see Section 12.4). In assessing the overall
evidence, the strengths and limitations of individual studies were evaluated based on
scientific considerations detailed in Table 12-5 of the Process Appendix
(Section 12.6.1). More details on the causal framework used to reach these conclusions
are included in the Preamble to the ISA (U.S. EPA. 2015). The evidence presented
throughout this chapter supports the following causality conclusions:

Outcome Group

Causality Determination

Pregnancy and Birth Outcomes

Likely to be Causal

Development

Causal

Female Reproductive Function

Likely to be Causal

Male Reproductive Function

Causal

The Executive Summary, Integrated Synthesis, and all other appendices of this Pb
ISA can be found at https://assessments.epa.gov/isa/document/&deid=359536.

8.1 Introduction and Summary of the 2013 Integrated Science
Assessment

This appendix evaluates the epidemiologic and toxicological literature related to the potential
effects of lead (Pb) on reproductive and developmental outcomes, divided into four sections: (1) effects
on pregnancy and birth outcomes; (2) effects on development; (3) effects on female reproductive
function; and (4) effects on male reproductive function. Based on the epidemiologic and toxicological
studies reviewed in the 2013 Pb Integrated Science Assessment (ISA) (U.S. EPA. 2013). the
determination for effects on pregnancy and birth outcomes was based on the mix of inconsistent results of
the epidemiologic and toxicological studies on various birth outcomes, but with some associations
observed in some epidemiologic studies of preterm birth and low birth weight and fetal growth. The

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determination for developmental effects was informed by evidence from toxicological studies reporting
delayed female sexual maturity and supported by epidemiologic studies of delayed pubertal onset for both
girls and boys. The determination for effects on female reproductive effects was based on epidemiologic
and toxicological studies for reproductive function among females reviewed including endpoints of
hormone levels, fertility, estrous cycle changes, and morphology or histology of female reproductive
organs including the placenta. Of the epidemiologic and toxicological studies reviewed for effects on
female reproductive function, the studies were high-quality and well-designed and examined different
exposure periods in conjunction with a number of outcomes related to female reproductive effects. The
determination for effects on male reproductive function were based on strong toxicological evidence that
showed detrimental effects on semen quality, sperm, and fecundity/fertility, with supporting evidence in
epidemiologic studies of associations between Pb exposure and detrimental effects on sperm. The
summary of the determinations from the 2013 Pb ISA is detailed below.

8.1.1	Effects on Pregnancy and Birth Outcomes

The 2013 Pb ISA (U.S. EPA. 2013) reported the associations between Pb exposure and birth
outcomes (infant mortality and embryogenesis; birth defects; preterm birth; and low birth weight/fetal
growth) were inconsistent overall. There were some associations observed between Pb and low birth
weight when epidemiologic studies used measures of postpartum maternal bone Pb or air exposures. The
associations were less consistent for maternal blood Pb measured during pregnancy or at delivery or
umbilical cord and placenta Pb (maternal blood Pb or umbilical cord and placenta Pb were the biomarkers
most commonly used in studies of low birth weight) but some associations between increased Pb
biomarker levels and decreased low birth weight/fetal growth were observed. Animal studies
investigating the effects of Pb exposure during gestation on litter size, implantation, and birth weight had
varying results between studies. Based on the mix of inconsistent results of studies on various birth
outcomes but some associations observed in select epidemiologic studies of preterm birth and low birth
weight/fetal growth, the evidence in the 2013 Pb ISA was suggestive of a causal relationship between Pb
exposure and birth outcomes.

8.1.2	Effects on Development

The 2013 Pb ISA (U.S. EPA. 20.1.3) reported Pb associated effects on development in
epidemiologic and toxicological studies. Previous toxicological studies indicated that delayed pubertal
onset may be one of the more sensitive developmental effects of Pb exposure with effects observed after
gestational exposures leading to blood Pb levels (BLLs) in the female pup of 1.3-13 (.ig/dL (lavicoli et aL
2006; lavicoli et aL 2004). Toxicological studies have reported delayed male sexual maturity as
measured with sex organ weight, seeing significant decrements at BLLs of 20-34 (.ig/dL (Ronis et ah.
1.998c; Sokol et aL. .1.985). The 2013 Pb ISA also presented findings from a toxicological study that

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suggests Pb may act through disruption of insulin-like growth factor 1 (IGF-1) to delay the onset of
puberty, demonstrated by the attenuation of Pb-induced delays in pubertal onset in female rats
supplemented with IGF-1 (Pine et ah. 2006'). Thus, data from the toxicological literature and from
epidemiologic studies demonstrated that puberty onset in both males and females is delayed with Pb
exposure. Findings from epidemiologic studies of the effect of Pb on postnatal growth were inconsistent
and findings from the toxicological literature of the effect of Pb exposure were mixed with recent growth
findings showing adult-onset male obesity after gestational and lactational Pb exposure. The 2013 Pb ISA
concluded that, based on the findings of delayed pubertal onset among males and females, there was
sufficient evidence to conclude a causal relationship between Pb exposure and developmental effects.

8.1.3	Effects on Female Reproductive Function

The 2013 Pb IS A (U.S. EPA. 2013) found some evidence of a potential relationship between Pb
exposure and female fertility; however, findings were inconsistent. Epidemiologic studies were largely
cross-sectional and adjustments for important confounding factors were not included in all studies. Some
toxicological studies reported effects on placental pathology and inflammation, decreased ovarian
antioxidant capacity, and altered hormone levels. Overall, the relationship observed with female
reproductive outcomes, such as fertility, placental pathology, and hormone levels in some epidemiologic
and toxicological studies was sufficient for the 2013 Pb ISA to conclude that evidence was suggestive of
a causal relationship between Pb exposure and female reproductive function.

8.1.4	Effects on Male Reproductive Function

The 2013 Pb ISA ("U.S. EPA. 2013) reported multiple studies in rodents and non-human primates
that observed Pb-induced sperm DNA damage, reduced sperm quality, reduced sperm production, and
histological and ultrastructural damage to male reproductive organs. Other toxicological studies reported
that Pb exposure was associated with decreases in reproductive organ weights, histological changes in the
testes and germ cell, and subfecundity. The 2013 Pb ISA also presented toxicological evidence suggesting
that Pb may damage sperm cells and sex organ tissue through induction of oxidative stress ("Salawu et ah.
2009; Shan et ah. 2009; Madhavi et ah. 2007; Rubio et ah. 2006; Wang et ah. 2006). Specifically, one
study reported Pb-induced increases in oxidative stress markers and reductions of levels of antioxidant
enzymes in testicular plasma ("Salawu et ah. 2009). In addition, several studies reported attenuation of Pb-
induced reductions in sperm count, motility, and viability when animals were co-administered substances
with known antioxidant properties (Salawu et ah. 2009; Shan et ah. 2009; Madhavi et ah. 2007; Rubio et
ah. 2006; Wang et ah. 2006). Epidemiologic studies were limited due to lack of consideration of potential
confounding factors or the use of men attending a fertility clinic, which could result in a biased sample.
However, a well-conducted epidemiologic study that enrolled men going to a clinic for either infertility
issues or to make a semen donation and controlled for other metals as well as smoking reported a positive

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association with various detrimental effects in sperm (Telisman et ah. 2001). Studies in the 2013 Pb ISA
that investigated the effects of Pb on hormone levels reported inconsistent results, resulting in uncertainty
as to whether Pb exerts its toxic effects on the reproductive system by affecting the responsiveness of the
hypothalamic-pituitary-gonad axis, by suppressing circulating hormone levels, or by some other pathway.
Based on the consistency and coherence of findings of the detrimental effects of Pb exposure on sperm
and semen in the toxicological literature from animal studies, the support from epidemiologic studies, and
biological plausibility provided by mode of action evidence; however, the evidence in the 2013 Pb ISA
was sufficient to conclude a causal relationship between Pb exposures and male reproductive function.

8.2 Scope

The scope of this section is defined by Population, Exposure, Comparison, Outcome, and Study
Design (PECOS) statements. The PECOS statements define the objectives of the review and establishes
study inclusion criteria thereby facilitating identification of the most relevant literature to inform the Pb
ISA.1 In order to identify the most relevant literature, the body of evidence from the 2013 Pb ISA was
considered in the development of the PECOS statements for this Appendix. Specifically, well-established
areas of research; gaps in the literature; and inherent uncertainties in specific populations, exposure
metrics, comparison groups, and study designs identified in the 2013 Pb ISA inform the scope of this
Appendix. The 2013 Pb ISA used different inclusion criteria than the current ISA, and the studies
referenced therein often do not meet the current PECOS criteria (e.g., due to higher or unreported
biomarker levels). Studies that were included in the 2013 Pb ISA, including many that do not meet the
current PECOS criteria, are discussed in this appendix to establish the state of the evidence prior to this
assessment. With exception of supporting evidence used to demonstrate the biological plausibility of Pb-
associated effects on reproductive and developmental health, studies evaluated and subsequently
discussed within this section were only included if they satisfied all the components of the following
discipline-specific PECOS statement:

Epidemiologic Studies:

Population: Any human population, including specific populations or lifestages that might be at
increased risk of a health effect.

Exposure: Exposure to Pb2 as indicated by biological measurements of Pb in the body - with a

'The following types of publications are generally considered to fall outside the scope and are not included in the
ISA: review articles (which typically present summaries or interpretations of existing studies rather than bringing
forward new information in the form of original research or new analyses), Pb poisoning studies or clinical reports
(e.g., involving accidental exposures to very high amounts of Pb described in clinical reports that may be extremely
unlikely to be experienced under ambient air exposure conditions), and risk or benefits analyses (e.g., that apply
concentration-response functions or effect estimates to exposure estimates for differing cases).

2Recent studies of occupational exposure to Pb were considered insofar as they addressed a topic area that was of
particular relevance to the National Ambient Air Quality Standards (NAAQS) review (e.g., longitudinal studies
designed to examine recent versus historical Pb exposure).

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specific focus on Pb in blood, bone, and teeth; validated environmental indicators of Pb
exposure;3 or intervention groups in randomized trials and quasi-experimental studies.

Comparison: Populations, population subgroups, or individuals with relatively higher versus
lower levels of the exposure metric (e.g., per unit or log unit increase in the exposure metric,
or categorical comparisons between different exposure metric quantiles).

Outcome: Reproductive effects, including but not limited to altered age of puberty onset, reduced
fertility, poor semen quality/motility, and miscarriage. Developmental effects including but
not limited to adverse pregnancy outcomes (e.g., reduced fetal growth, preterm birth, small
for gestational age [SGA], birth defects), as well as postnatal developmental effects.

Study Design: Epidemiologic studies consisting of longitudinal and retrospective cohort studies,
case-control studies, cross-sectional studies with appropriate timing of exposure for the health
endpoint of interest, randomized trials and quasi-experimental studies examining
interventions to reduce exposures.

Experimental Studies:

Population: Laboratory nonhuman mammalian animal species (i.e., mouse, rat, Guinea pig,

minipig, rabbit, cat, dog; whole organism) of any lifestage (including preconception, in utero,
lactation, peripubertal, and adult stages).

Exposure: Oral, inhalation, or intravenous routes administered to a whole animal (in vivo) that
results in a BLL of 30 (ig/dL or below.4,5

Comparators: A concurrent control group exposed to vehicle-only treatment or untreated
control.

Outcomes: Reproductive and developmental effects.

Study design: Controlled exposure studies of animals in vivo.

8.3 Effects on Pregnancy and Birth Outcomes

The 2013 Pb ISA reported inconsistent findings in the epidemiologic and toxicological literature
for birth outcomes (infant mortality and embryogenesis; birth defects; preterm birth; and low birth
weight/fetal growth). Among the epidemiologic studies, there were inconsistent associations between Pb

3Studies that estimate Pb exposure by measuring Pb concentrations in PMio and PM2.5 ambient air samples are only
considered for inclusion if they also include a relevant biomarker of exposure. Given that size distribution data for
Pb-PM are fairly limited, it is difficult to assess the representativeness of these concentrations to population
exposure |Section 2.5.3 (U.S. EPA. 2013YL Moreover, data illustrating the relationships of Pb-PM n, and Pb-PM; >
with BLLs are lacking.

4Pb mixture studies are included if they employ an experimental arm that involves exposure to Pb alone.

5This level represents an order of magnitude above the upper end of the distribution of U.S. young children's BLLs.
The 95th percentile of the 2011-2016 National Health and Nutrition Examination Survey (NHANES) distribution of
BLL in children (1-5 years; n = 2,321) is 2.66 (ig/dL (CDC, 2019) and the proportion of individuals with BLLs that
exceed this concentration varies depending on factors including (but not limited to) housing age, geographic region,
and a child's age, sex and nutritional status.

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exposure and preterm birth. A single study of neural tube defects (NTDs) found no associations in the
2013 Pb ISA. but studies within the 2006 Air Quality Criteria Document for Lead (Pb AQCD) (U.S. EPA.
2006') reported associations between Pb exposure and NTDs. There were some associations reported
between Pb and low birth weight when epidemiologic studies used measures of postpartum maternal bone
Pb or air exposures. There were less consistent associations for maternal blood Pb measured during
pregnancy or at delivery or umbilical cord and placenta Pb (maternal blood Pb or umbilical cord and
placenta Pb were the biomarkers most commonly used in studies of low birth weight). The effects of Pb
exposure during gestation in animal toxicological studies included mixed findings, but most studies
reported reductions in birth weight of pups or birth weight of litters when dams were treated with Pb.

The recent epidemiologic and toxicological studies are detailed in the following sections. Effects
on pregnancy and birth outcomes encompass a large range of outcomes. The following sections relating to
pregnancy and outcomes are categorized into seven main sections: (1) maternal health during pregnancy;
(2) prenatal growth; (3) preterm birth; (4) birth defects; (5) spontaneous abortion and pregnancy loss and
fetal and infant mortality; (6) placental function; and (7) other pregnancy and birth outcomes.

8.3.1 Maternal Health During Pregnancy

Maternal health during pregnancy encompasses a wide range of health effects. The details of the
recent epidemiologic and toxicological studies evaluating the association between Pb exposure and
maternal health during pregnancy are provided in Table 8-2 and Table 8-3, respectively.

8.3.1.1 Epidemiologic Studies on Maternal Health During Pregnancy

The main maternal health outcomes evaluated in this section are gestational diabetes mellitus
(GDM) and epigenetic studies. Although there are a limited number of epigenetic studies, these studies
may help to add support for biological plausible pathways for which Pb exposure may affect maternal
health during pregnancy.

8.3.1.1.1	Epidemiologic Studies on Gestational Diabetes Mellitus

There were no studies on GDM in the 2013 Pb ISA. There were several recent epidemiologic
studies that evaluated the association between Pb exposure and GDM and/or impaired glucose tolerance
(IGT) ("Tatsuta et aL 2022a; Zheng et aL 2021; Zhou et aL 2021b; Qguri et ah. 20.1.9; Soomro et al..
20.1.9; Wang et al.. 20.1.9; Shapiro et al.. 2015). Generally, across the studies there were null associations
between Pb exposure and GDM and IGT, and/or GDM or IGT. In studies that evaluated Pb in maternal
blood with GDM outcomes, the timing of when Pb was measured differed between trimesters, but the
difference in what trimester Pb was measured did not impact the associations ("Qguri et al.. 20.1.9; Soomro

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et ah. 20.1.9; Wang et ah. 20.1.9; Shapiro et ah. 2015). These studies all reported median BLLs less than
5 (ig/dL (range: 1.7-2.8 (ig/dL) or geometric mean BLLs less than 5 (ig/dL (range: 0.6-1.62 (ig/dL or
6.05-6.13 ng/g). Additionally, while maternal blood was the primary biomarker used to measure Pb
exposure, some studies have used other biomarkers such as maternal serum (Zhou et ah. 2021b') and
maternal erythrocytes ("Zheng et ah. 2021); however, the type of biomarker measurement did not
influence the pattern of associations. Only one study reported a decrease of 0.5 (95% confidence interval
[CI]: -1.6, -0.6) mg/dL difference in mid-gestational glucose concentration associated with an
interquartile range (IQR) (17.6 ng/g) change in blood erythrocyte Pb exposure (Zheng et ah. 2021).
Furthermore, multiple studies also considered co-exposure to other metals in addition to Pb, but the
associations remained null (Zheng et ah. 2021; Zhou et ah. 2021b; Qguri et ah. 20.1.9; Wang et ah. 20.1.9).
Overall, the associations between Pb exposure and GDM, IGT, and GDM or IGT were null, and the null
associations persisted across the different trimesters of when Pb levels were measured, the different
biomarkers for Pb exposure, and adjustment for co-exposure to other metals.

8.3.1.1.2	Epidemiologic Studies on Epigenetic Effects During Pregnancy

There were no studies on epigenetic effects during pregnancy evaluated in the 2013 Pb ISA. The
recent epidemiologic studies on epigenetic effects during pregnancy are limited but provide insight on
potential mechanistic pathways in which Pb exposure may impact pregnancy. A single study by Sanders
et ah (20.1.5) assessed the association between maternal Pb levels in blood, patella, and tibial bone and
altered micro RNA (miRNA) expression in the cervix during the second trimester of pregnancy in a
subset of 60 women enrolled in a prospective birth cohort, Programming Research in Obesity, Growth,
Environment and Social Stressors (PROGRESS), in Mexico City. Changes in cervical miRNA expression
are a potential mechanism that could alter gene expression leading to aberrant changes in cervix tissue
function and subsequently impact parturition (Sanders et ah. 20.1.5). Expression of certain miRNAs in the
cervix during pregnancy have been associated with subsequent gestational age (GA) at delivery ("Sanders
et ah. 20.1.5). During mid-pregnancy (16-19 weeks gestation), samples from cervical exams were
collected and analyzed for the expression profiles of 800 miRNAs. Overall, there were distinct miRNAs
measured in cervical samples during pregnancy that are associated with the subsequent GA of offspring.
Sanders et ah (20.1.5) also identified differentially expressed miRNAs with respect to preterm compared
term birth in a subset of women. There were two miRNAs expressed in the cervix that were identified in
association with maternal second trimester BLLs, seven miRNAs that were identified in association with
maternal patella bone Pb levels, and six miRNAs that were identified in association with maternal tibia Pb
levels (see Table 8-2). In another epigenetics study in the same PROGRESS cohort, Sanchez-Guerra et ah
(20.1.9) assessed the association of blood Pb exposure during pregnancy with mitochondrial DNA
(mtDNA) content, which is a sensitive marker of mitochondrial function and oxidative stress, in cord
blood. Maternal blood Pb samples were obtained at three time points (second trimester n = 410, third
trimester n = 356, and at delivery n = 354), and cord blood (n = 346) Pb samples were obtained at
delivery. Maternal Pb levels during the second trimester ((3: 0.017 [95% CI: 0.002, 0.031]) were

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associated with higher mtDNA content; however, there were null associations between cord BLLs at
delivery ((3: 0.016 [95% CI: 0.001, 0.03]), maternal third trimester blood Pb ((3: 0.015 [95% CI 0.00,
0.03]), and maternal BLLs at delivery ([3: 0.013 [95% CI: -0.001, 0.027]). These epigenetic studies
provide support of potential mechanistic pathways in which Pb exposure is associated with maternal
health during pregnancy.

8.3.1.1.3	Epidemiologic Studies on Other Outcomes Related to Maternal Health During

Pregnancy

There were several other outcomes related to maternal health during pregnancy. More specific
study details, including Pb levels, study population characteristics, potential confounders, and select
results from these studies are highlighted in Table 8-2. In other outcomes related to maternal health
during pregnancy, Pb exposure has been associated with decreased free thyroxine (FT4) during mid-
pregnancy ("Kahn et ah. 2014); increased thyroid peroxidase antibodies (TPOAb) during mid-pregnancy
("Kahn et ah. 20.1.4); small increases in umbilical cord blood Pb and elevations in systolic blood pressure
and diastolic blood pressure during labor and delivery (Wells et ah. 2011); changes in Global Severity
Index (GSI), depression and anxiety symptom scores (Li et ah. 20.1.7b); bone mineral density of the
patella (Osorio-Yanez et ah. 2021); increased matrix metalloproteinases (MMP), regulators of uterine
remodeling (Kim et ah. 2022); and increased risk of preeclampsia (Gaiewska et ah. 2021; Wti et ah.

2021).	However, there were no associations between Pb exposure and reduced Cortisol awakening
response (Braun et ah. 20.1.4); maternal depression (Ishitsuka et ah. 2020); anti-Miillerian hormone
(AMH), a suggested marker of ovarian function and biological marker of female fecundity (Christensen et
ah. 20.1.6); hormone levels in pregnancy (Gustin et ah. 2021); or thyroid function (Corrales Vargas et ah.

2022).

8.3.1.2 Toxicological Studies on Maternal Health During Pregnancy

Previous Pb ISAs and AQCDs did not report any toxicological studies that investigated the
effects of Pb on maternal health during pregnancy. Despite this lack of prior studies to compare to, recent
toxicological studies have reported on the effects of Pb on maternal weight gain during pregnancy
(Table 8-3). Maternal weight gain is often used as an indicator of fetal growth and maternal overt toxicity.
Additionally, maternal weight gain shares associations with gestational conditions in humans (Santos et
ah. 201.9). Recent studies dosed Sprague-Dawley rats with Pb viagavage for the first 20 days of
pregnancy and reported that the 160 ppm Pb treatment group exhibited reduced weight gain during
pregnancy (maternal BLLs on gestational day |GD| 20 were reported to be 23.9-27.7 (.ig/dL) (Saleh et ah.
20.1.9; Sal eh et ah. 20.1.8). Of note is that both studies by Saleh et al. (Sal eh et ah. 20.1.9; Sal eh et ah. 20.1.8)
reported reduced brain weight of dams, indicating that overt toxicity may have contributed to the overall
reduction in maternal weight. Further, the reported maternal BLLs were higher than those observed in the

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following studies that investigated the same outcomes. Corv-Slechta et al. (20.1.3) and Schneider et al.
i2 both dosed C57BL/6 mice with 100 ppm Pb via drinking water starting 2 months prior to mating
and reported no effects on maternal bodyweight gain and observed much lower maternal BLLs with
Corv-Slechta et al. (20.1.3) reporting 12.12 (.ig/dL at weaning and Schneider et al. (20.1.6) reporting
12.61 (ig/dL on lactation day 21. Similarly, Wang et al. (20.1.4) reported no effects in Wistar rats dosed
with Pb via drinking water for various durations during pregnancy. In Wang et al. (2014) dosing from
GD 1-10, GD 11-20, and GD 1-20 resulted in maternal BLLs of 26.4, 12.4, and 36.0 (ig/dL, respectively,
at termination of the study on GD 20. Although some of these BLLs overlap with those seen in the studies
by Saleh et al. (2018) and Sal eh c	).1.9) wherein suppression of maternal weight gain was observed,

it is possible that the use of different strains or different dosing routes could be attributed to the observed
difference in effect on maternal weight gain. Additionally, the reduction of brain weight observed in the
dams used in the studies by Saleh et al. (2018) and Saleh etal. (20.1.9) suggest that maternal overt toxicity
may be responsible for the observed reduction in maternal weight gain.

8.3.1.3 Integrated Summary of Effects on Maternal Health During Pregnancy

The 2013 Pb ISA did not include epidemiologic and/or toxicological studies that evaluated the
relationship between Pb exposure and maternal health during pregnancy. There were consistent null
associations between Pb exposure and GDM among the recent epidemiologic studies. While the critical
window for GDM is unknown, these studies had different time points during pregnancy in which Pb
exposure was measured and different biomarkers of exposure (blood, serum, and erythrocyte) and the null
associations persisted. A few of the studies were limited by the cross-sectional study design and the small
number of GDM cases. Additionally, a few of the recent epidemiologic studies incorporated mixture
methods to consider Pb exposure in conjunction with co-exposure to other metals to evaluate associations
with GDM, which helps to reduce uncertainties regarding co-pollutant confounding. The limited number
of epigenetic studies provide support of potential mechanistic pathways in which Pb exposure are
associated with selected maternal health during pregnancy. Furthermore, there was a small body of
evidence across various additional pregnancy-related endpoints in the epidemiologic literature; however,
the small number limits the ability to judge coherence and consistency across these studies, although the
positive associations observed demonstrate that Pb exposure could result in physiological responses that
contribute to adverse pregnancy outcomes (e.g., changes in thyroid function, maternal mental health,
changes in blood pressure, preeclampsia). In the recent toxicological literature, there were a limited
number of studies that investigated the relationship between Pb exposure and maternal weight gain during
pregnancy; however, the only studies that observed changes in maternal weight gain also reported signs of
possible overt toxicity (reduced brain weight), indicating that weight gain during pregnancy may not have
been a direct effect of Pb exposure. The majority of recent toxicological studies in rodents reported that
maternal weight gain during pregnancy was unaffected by Pb exposure.

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8.3.2

Prenatal Growth

The recent epidemiologic and toxicological studies that examined the relationship between Pb
exposure and prenatal growth, which includes outcomes such as fetal growth, birth weight, body length at
birth, and GA, are summarized in the text below. Study details of the recent epidemiologic studies are
included in Table 8-4 and the recent toxicological studies are in Table 8-3.

8.3.2.1 Epidemiologic Studies on Prenatal Growth

The epidemiologic studies in the 2013 Pb ISA reported associations between maternal bone Pb
and low birth weight and with studies of Pb air exposures and birth weight. The associations were less
consistent when using maternal blood Pb or umbilical cord and placenta Pb as the exposure measurement,
although some studies did demonstrate associations. The studies of Pb exposure and fetal growth were
limited by their cross-sectional study design, small sample size, high air Pb concentrations (air Pb as high
as 30 |ig/m3). and in some studies, the lack of control of confounders.

A large number of epidemiologic studies have been published since the 2013 Pb ISA on exposure
to Pb and prenatal growth. The studies in this section focus on these prenatal growth outcomes, including
birth weight; low birth weight; body length, crown-to-heel length, head circumference (HC), Ponderal
Index (PI; weight/height3), GA, SGA, and large for gestational age (LGA). Multiple cross-sectional and
cohort studies have been conducted that examined the relationship between Pb exposure and prenatal
growth; however, the findings from the recent epidemiologic studies are inconsistent. There are
differences in study design, timing of the exposure (at different points during pregnancy, at delivery),
differences in biomarkers examined for Pb (maternal blood, maternal serum, cord blood, maternal red
blood cells, placental tissue), and small sample sizes in some studies. The study details, including
information on study population, biomarker of exposure, and outcome, are in Table 8-4.

Several studies used cord blood to assess Pb exposure and reported null associations with birth
weight (Lee et ah. 2021; Govarts et ah. 2020; Tatsutaet ah. 20.1.7; Wang et ah. 20.1.7b; Govarts et ah.
20.1.6; Garcia-Esauinas et ah. 20.1.3; Xie et ah. 2013). while a single study reported a reduction in birth
weight (Xu et ah. 20.1.2). Among these studies, there were also inconsistent associations when examining
cord blood Pb exposure and birth weight among infant sex. Tatsutaet ah (20.1.7) evaluated the
associations between cord blood Pb and birth weight between male and female infants, but the
associations remained null. While there were null associations with birth weight, birth length, HC, and PI
when infant sexes were analyzed together, analyses stratifying by infant sex reported associations in male
infants, including increased birth weight ((3: 206.50 [95% CI: 46.15, 366.86]) and decreased HC ((3: -0.65
[95% CI: -1.24, -0.06]) per 1-unit increase logio-Pb cord blood concentration. Among female infants,
there was only a reduction in PI ([3: -0.16 [95% CI: -0.30, -0.02]) per 1-unit increase in the logio-Pb cord
blood concentration (Wang et ah. 2017b).

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In addition to birth weight, there were several other prenatal growth outcomes in these studies
that were evaluated in association with cord blood Pb; however, the associations were inconsistent.
Garcia-Esauinas et al. (20.1.3) also reported null associations between cord blood Pb and birth length, and
1- and 5-minute Apgar scores from the 144 newborns who were a part of cross-sectional biomonitoring
study of the BioMadrid Project. Xu et al. (20.1.2) also reported decreased mean GA of 0.57 weeks (95%
CI: 0.51, 0.63), with increased risk of low birth weight rate (OR: 1.61 [95% CI: 1.37, 1.90]), and
increased risk of intrauterine growth retardation rate (OR: 2.12 [95% CI: 1.68, 2.69]). Xie et al. (20.1.3)
reported a negative association with birth length ([3: -0.84 cm [95% CI: -1.52, -0.16]) per square root 1-
(ig/dL increase in cord blood Pb, but null associations with birth weight ([3: -99.33 g [95% CI: -217.33,
20.67]) and HC ([3: -0.36 [95% CI: -0.81, 0.03]). A single study that was conducted among 1,578
mother-infant pairs in Saudi Arabia reported no associations between cord BLLs and PI below the 10th
percentile (OR: 0.66 [95% CI: 0.42, 1.05]) (Al-Saleh etal.. 2014).

Maternal blood was also used to measure Pb exposure in association with prenatal growth
outcomes in multiple cross-sectional studies, but the associations were inconsistent. Xie et al. (20.1.3)
reported a negative association with birth weight ([3: -148.99g [95% CI: -286.33,-11.66]) per square
root 1-jxg/dL increase in maternal blood Pb measured at delivery, but null associations with birth length
([3: -0.46 cm [95% CI: -1.25, 0.34]) and HC ([3: -0.37 cm [95% CI: -0.78, 0.19]) among 252 mother-
infant pairs in a rural area located on the south coast of Laizhou Bay, China between 2010 and 2011.
However, Kim et	10) reported negative associations between maternal blood natural log (ln)-Pb.

measured at delivery, and HC ([3: -0.75 cm [95% CI: -1.17, -0.32]) and PI ([3: -0.62 kg/m3 [95% CI:
-1.13, -0.11]), but there were null associations with birth weight ([3: 60 g [95% CI: -15, 135]), BMI ([3:
-0.14 kg/m2 [95% CI: -0.39, 0.11]), and SGA (OR: 0.69 [95% CI: 0.33, 1.46]) among participants of e-
waste Recycling Exposure and Community Health (e-REACH) Study. A study by Xu et	*2b)

reported that a one ln-unit increase in maternal BLLs, measured at delivery, was associated with increased
GA ([3: 0.18 weeks [95% CI: 0.05, 0.31]), decreased birth length ([3: -0.39 cm [95% CI: -0.66, -0.22]),
and decreased HC ([3: -0.22 cm [95% CI: -0.39, -0.06]), but a null association with birth weight. There
were also null associations across tertiles of maternal BLLs and low birth weight.

In addition to cord blood and maternal blood, other biomarkers such as maternal serum, cord
blood serum, and placental tissue were used to assess Pb exposure with birth weight among other studies
and reported inconsistent associations ("Yang et al.. 2020: Freire et al.. 20.1.9: Mikelson et al.. 20.1.9: Tang
et al.. 20.1.6: Hu et al.. 20.1.5). A study that measured Pb in both maternal serum, measured at delivery, and
cord blood serum reported null associations with birth weight for both biomarkers (Hu et al.. 20.1.5). while
another study reported null associations between cord blood serum and birth weight-for-gestational-age
Z-score. when modeled continuously or categorized by quintiles (Yang et al.. 2020). Although there were
null associations with birth weight and GA, there was a decrease in birth height and a decrease in HC per
ln-Pb increase in umbilical cord serum among 103 mother-newborn pairs from an island in the East China
Sea (Tang et al.. 20.1.6). When placental tissue was the biomarker of exposure for Pb, a single cross-
sectional study reported null associations with birth weight, low birth weight, birth, head, GA, and SGA

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(Freire et a!.. 2019). but another reported a decrease in birth weight of 58.3 g (95% CI: -97.9, -18.8) per
ln-Pb increase in placental tissue (Mikelson et ah. 20.1.9).

The use of advanced statistical methods to evaluate the impact of co-exposure to other metals, or
mixtures, helps to address uncertainties of co-pollutant confounding. To assess the associations between
metal mixtures (arsenic [As], cadmium [Cd], manganese [Mn], and Pb) in umbilical cord blood and birth
weight, birth length, and HC, 1,088 participants of a birth cohort in Bangladesh were assessed in a cohort
study (Lee et ah. 2021). There were null associations with birth weight ((3: -0.04 g [95% CI: -0.19,
0.11]), birth length ([3: -0.06 cm [95% CI: -0.20, 0.09]), and HC ([3: 0.08 cm [95% CI: -0.06, 0.23]) in
association with an IQR increase in ln-Pb cord blood concentrations, when adjusted for confounders and
other metals. In addition to the multivariable regression analysis, Lee et ah (202.1.) also used Bayesian
kernel machine regression (BKMR) to estimate the effects of co-exposure to metal mixtures. BKMR is a
method that estimates the multivariable exposure-response function in a flexible and parsimonious way,
conducts variable selection on the (potentially high-dimensional) vector of exposures, and allows for a
grouped variable selection approach that can accommodate highly correlated exposures. In the BKMR
analysis, there was an inverse association between the metal mixture overall and birth length when all
four metal concentrations were >60th percentile and HC when all four metals were >5 5th percentile,
compared to their median values, with stronger associations as the concentrations of the four metals
increased. However, when estimating the difference in birth size with an IQR increase in each individual
metal when the other metals were fixed at their 25th, 50th, or 75th percentiles, the associations with Pb
were null.

Overall, in the multiple longitudinal birth cohort studies, there were inconsistent findings between
various Pb exposure biomarkers and prenatal growth outcomes. The multiple longitudinal birth cohort
studies have reported inconsistent associations. These studies collected maternal samples during different
time periods during pregnancy and utilized different biomarkers to measure Pb exposure to evaluate
associations with a variety of prenatal growth outcomes. In the longitudinal studies that measured Pb
exposure from maternal blood, there were inconsistent patterns of association with prenatal growth
outcomes, regardless of the trimester Pb exposure was measured or prenatal growth outcome (see
Table 8-4). Several studies reported null associations with birth weight (Shih et ah. 2021; Woods et ah.
20.1.7; Taylor et ah. 20.1.6; Bloom et ah. 20.1.5; Garcia-Esquinas et ah. 20.1.4; Rabito et ah. 20.1.4) and birth
weight Z-score (BWZ) (Daniali et ah. 2023). while others reported reductions in birth weight (Goto et ah.
2021; Hu et ah. 20. I. L tdosthenous et ah. 20.1.7; Taylor et ah. 20.1.5). Of note, Rodosthenous et ah (. 01
measured Pb levels in maternal blood during the second trimester among 944 mother-infant pairs in the
PROGRESS cohort in association with birth weight using both linear and quantile regression. While the
linear regression reported a null association with birth weight-for-gestational-age Z-score ([3: -0.06 [95%
CI: -0.13, 0.003]) per log2-Pb blood level increase, the quantile regression analysis revealed larger
magnitudes of the association maternal blood Pb and birth weight-for-gestational-age Z-score. The
magnitude of the association was largest in the lowest (<30th) Z-score percentiles (difference in Z-score

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ranged from -0.13 to -0.08). The use of quantile regression provides insights to potential sensitivity to Pb
exposure for smaller infants, an association that was not detected by linear regression.

While some studies reported null associations with birth length (Daniali et ah. 2023; Shih et aL
2021; Bloom et aL 2015). a single study reported a 0.20 cm decrease (95% CI: -0.30, -0.10) in birth
length per 1 (ig/dL increase in maternal BLL (collected during the second or third trimester) among
participants of the Japan Environment and Children's Study (J ECS) (Goto et aL. 2021). A single study
reported a reduction in HC of 0.03 cm (95% CI -0.06, -0.00) per 1 (ig/dL increase of first trimester
maternal blood Pb ("Taylor et al.. 20.16). but other studies reported null associations with HC and first
trimester maternal blood Pb (Daniali et al.. 2023; Taylor et al.. 20.1.5). pre-pregnancy maternal and
parental blood (Bloom et al.. 2015). or second or third trimester maternal blood Pb (Shih et aL. 202.1.).
While there was reported decreased GA ((3: -1.9 days [95% CI: -3.1, -0.5]) per IQR increase in second
trimester maternal ln-Pb blood level among those in Puerto Rico Testsite for Exploring Contamination
Threats (PROTECT) cohort (Ashrap et al.. 2020). there were null associations with gestational and pre-
pregnancy maternal and parental blood (Bloom et al.. 20.1.5) and maternal BLLs during the second or third
trimester among the JECS (Goto et al.. 2021); however, Goto et al. (202.1.) did report an increased risk of
SGA (OR: 1.34 [95% CI: 1.16, 1.55]) and increased risk of low birth weight (OR: 1.34 [95% CI: 1.16,
1.55]) per 1 (ig/dL increase in maternal BLL, but other studies did not report increased risk of SGA
(Thomas et al.. 20.1.5) and maternal blood (collected during the first and third trimesters of pregnancy) or
second trimester maternal blood (Ashrap et al.. 2020). There were consistent null associations with PI and
maternal blood (Shih et al.. 2021; Bloom et al.. 20.1.5) and crown-to-heel length and first trimester
maternal blood Pb (Taylor et al.. 20.1.6; Taylor et al.. 20.1.5).

In addition, some of the longitudinal studies considered different effect modifiers when assessing
the associations between maternal BLLs and prenatal growth outcomes. Among participants in the
Canadian Maternal-Infant Research on Environmental Chemicals (MIREC) study, there were null
associations between maternal blood (collected during the first and third trimesters of pregnancy) and
SGA across tertiles of maternal blood Pb (Thomas et al.. 20.1.5). In addition, an exploratory analysis was
conducted to examine the potential effect modification of single nucleotide polymorphisms (SNP) in
GSTP1 and GSTOl genes on the relationship of maternal blood Pb and SGA. There was a marginal
interaction between maternal Pb exposure and the GSTP1 Al 14V SNP (p = 0.06), but there was no
indication of effect modification by other GSTP1 and GSTO1 SNPs on the associations between maternal
blood Pb and SGA. In another study in the PROTECT cohort, the modifying effect of psychosocial stress
on the association between maternal blood Pb exposure and GA, BWZ, SGA, and LGA were examined in
a subset of 682 pregnant women (Ashrap et al.. 2021). Maternal blood samples were collected at
18 ± 2 weeks gestation and 26 ± 2 weeks gestation. Among mothers who reported "good" psychosocial
status, there was decreased gestation age ([3: -1.9 days [95% CI: -3.2, -0.6]); however, there were null
associations with BWZ ([3: 0.1 [95% CI: 0.0, 0.2]), SGA (OR: 0.86 [95% CI: 0.65, 1.14]), and LGA (OR:
0.89 [95% CI: 0.64, 1.23]). The associations for mothers who reported "poor" psychosocial status were
null across the birth outcomes.

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In addition to the associations between prenatal growth outcomes and Pb levels, there were sex-
stratified differences. In a study by Garcia-Esquinas et al. (20.1.4). 97 mother-father-infants in the
BioMadrid Study were used to evaluate associations between prenatal Pb exposure and fetal development
from three biomarkers (maternal and paternal blood Pb at 34 weeks gestation and cord blood at delivery)
with different growth metrics at birth. While there were no associations between log-Pb blood levels
(maternal, paternal, or cord) and gestation age, birth weight, birth length, abdominal diameter, or cephalic
diameter (CD), associations were observed when analyses were stratified by infant sex. Among female
infants, there was decreased birth length of 1.06 cm (95% CI: -2.03, -0.08) and CD of-0.55 cm (95%
CI: -1.03, -0.07) per two-fold increase in paternal BLLs (|ig/L). but there were no associations among
male infants. In the study by Sfaih et al. (2021). there were null associations between maternal blood log;-
Pb concentrations (collected between 6 and 32 weeks of gestation) and prenatal growth outcomes (GA,
birth weight, birth length, HC, and PI). However, when stratified by infant sex, there were reductions in
GA ((3: -0.98 weeks [95% CI: -1.67, -0.30]), birth weight ((3: -381 g [95% CI: -583, -178]), birth length
([3: -1.44 cm [95% CI: -2.45,-0.42]), and HC ([3: -1.10 cm [95% CI: -1.70, -0.50]), but had a null
association with PI ([3: -1.07 kg/m3 [95% CI: -1.56, 0.39]) among female infants, while the associations
for these same outcomes were null among male infants.

There were also a limited number of studies that considered co-exposure to other pollutants. From
the MIREC study, 1,857 mother-infant pairs were analyzed to examine the relationship between prenatal
exposure to a mixture of endocrine-disrupting chemicals, including Pb, and birth weight using BKMR
(H.u et al.. 2021). Maternal blood was collected during the first trimester of pregnancy. In the adjusted
model for log2-Pb, every two-fold increase in Pb concentration was associated with a mean birth weight
reduction of 82.22 g (95%: -145.46, -18.97), and when adjusted for other metals, the reduction in mean
birth weight was 75.89 g (95% CI: -141.24, -10.54). In the mixtures analysis, Pb was the main
contributor to the adverse effect on birth weight in the metal mixture consisting of As, Cd, mercury (Hg),
Mn, and Pb. An increase in the log2-Pb concentration from the 25th to the 75th percentile was associated
with a posterior mean of -47g, meaning that there was a reduction in mean birth weight of 47 g, while
holding the other components in the metal mixture constant at their median values.

In addition to maternal blood Pb, other biomarkers such as maternal erythrocytes, maternal
serum, and teeth were used to assess Pb exposure with prenatal birth outcomes, including birth weight,
birth length, or HC. Maternal erythrocytes from blood samples were collected during the third trimester
(mean: gestational week 29) from 584 mothers in the Nutritional impact on Immunological maturation
during Childhood in relation to the Environment (NICE) study in Northern Sweden (Gustin et al.. 2020).
Maternal erythrocytes reflect exposure over the past 1-3 months. A doubling of maternal erythrocyte Pb
concentration was not associated with birth weight ([3: -13 g [95% CI: -66, 41]), birth length ([3:
-0.080 cm [95% CI; -0.31, 0.15]), or HC ([3: 0.059 cm [95% CI: -0.22, 0.34] for maternal erythrocyte Pb
concentration less than the median of 14 |ig/kg and [3: -0.24 cm [95% CI: -0.53, 0.056] for maternal
erythrocyte Pb concentration greater than median of 14 |ig/kg). There was no interaction by infant sex.
When mutually adjusted for other maternal metal exposure to Cd and Hg, the null associations persisted.

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In a subset of the Project Viva prospective pre-birth cohort, individual and joint effects of metal mixture
components on birth weight, length, HC, and GA were estimated in association with maternal erythrocyte
Pb concentrations collected during early pregnancy (11.3 ± 2.8 weeks of gestation) from 1,423 mother-
infant pairs (Rahman et ah. 2021). In single metal model, an IQR increase in maternal erythrocyte Pb
concentration was associated with a 33.9 g (95% CI: -65.3, -2.5) decrease in birth weight, but there were
no associations with birth length ((3: -0.10 cm [95% CI: -0.29, 0.09]), HC ((3: -0.07 cm [95% CI: -0.17,
0.04]), or GA ([3: 0.03 weeks [95% CI: -0.10, 0.16]). When stratified by infant sex, the associations were
null for both male and female infants and birth weight, birth length, HC, and GA. Additionally, there was
consistent pattern of association of decreased birth weight, birth length, and HC overall and in the infant
sex-stratified analyses (see Table 8-4).

A total of 3,125 mother-infant pairs were recruited from the China-Anhui Birth Cohort Study
(C-ABCS) to investigate the associations between maternal serum Pb levels the first trimester (median of
11 weeks gestation) and in the second trimester (median of 16 weeks gestation) with growth metrics
(Wang et aL 2017a). Overall maternal serum Pb during pregnancy had a negative association with birth
weight ([3: -2.74 g [95% CI: -5.17,-0.31]), but null associations with birth length, HC, and chest
circumference. When stratified by trimester, the negative association with birth weight persisted, with a
reduction of 4.40 g (95% CI: -8.22, -0.58) for first trimester maternal serum Pb and a 1.64 g (95% CI:
-4.80, -0.58) reduction for second trimester maternal serum Pb. There were no associations by trimester
maternal serum Pb for birth length, HC, or chest circumference. In addition, there was increased risk of
SGA of 1.45 (95% CI: 1.04, 2.02) for subjects with medium-Pb maternal serum (1.18-1.70 (ig/dL) and
increased risk of SGA of 1.69 (95% CI: 1.22, 2.34) in subjects with high-Pb maternal serum
(>1.71 (ig/dL), compared to low-Pb maternal serum (<1.18 (.ig/dL). When stratified by infant sex, there
was increased risk of SGA among female infants (OR: 1.51 [95% CI: 0.99, 2.31] for medium-Pb maternal
serum and OR: 1.68 [95% CI: 1.12, 2.54] for high-Pb maternal serum), but among male infants, the
associations were null. There was an increased risk of SGA with high first trimester maternal serum Pb
(OR: 2.13 [95% CI: 1.24, 3.38]), but there were null associations among second trimester maternal serum
Pb.

In a small cohort study, second and third trimester Pb levels were estimated from baby teeth from
145 participants in the Wayne County Health, Environment, Allergy and Asthma Longitudinal Study
(WHEALS) (Cassidv-Bushrow et aL 20.1.9). There were no associations between tooth Pb in the second
or third trimester and BWZ ([3: -0.15 [95% CI: -0.35, 0.05] for second trimester and [3: -0.06 [95% CI:
-0.24, 0.12] for third trimester) or GA at delivery ([3: 0.08 [95% CI: -0.19, 0.35] for second trimester and
[3: 0.14 [95% CI: -0.11, 0.39] for third trimester) in the fully adjusted models. There was no indication
that there was a time effect (difference between the effect estimates in the second and third trimesters) for
birth weight for Z-score ([3: -0.31 [95% CI: -0.90, 0.28]) or GA at delivery ([3: -0.22 [95% CI: -1.08,
0.64]). Additionally, when stratified by child's sex, there were no associations between tooth Pb in the
second or third trimester and BWZ or GA at delivery.

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In a study by Bui et al. (2022). effects of short-term maternal exposure to airborne Pb during
pregnancy on birth weight, low birth weight, and SGA was estimated using a quasi-experimental variation
in airborne Pb exposure based on the National Association for Stock Car Auto Racing (NASCAR)'s
deleading of racing fuel in a difference-in-difference model in the Charlotte-Concord-Gastonia
Metropolitan Statistical Area in North Carolina. After deleading of racing fuel, there was an average
increase in birth weight of 102.50 g (95% CI: 45.73, 159.2), decreased probability of low birth weight of
0.0445 (95% CI: -0.0697, -0.0194), and reduction in the probability of SGA of 0.0396 (95% CI:
-0.0638, -0.0155) among children born to mothers residing less than 4000 meters of the Charlotte Motor
Speedway, compared with those residing greater than 10,000 meters. The difference-in-difference
methodology allows for the control of time-varying confounders, removing biases from comparisons over
time in the treatment group that could be the result of trends due to other causes of the outcome.

8.3.2.2	Toxicological Studies on Prenatal Growth

The 2013 Pb ISA discussed a few studies that reported reduced birth weight of offspring from Pb-
treated dams (Masso-Gonzalez and Antonio-Garcia. 2009; Wang et al.. 2009; Teiion et al.. 2006). Recent
toxicological studies consistently report no effects of Pb on birth weight (Table 8-3). Most studies began
exposure of the dam prior to conception of the offspring (Zhao et al.. 2021; Tartaglione et al.. 2020; Rao

Barkur and Bairv. 20.1.6; Schneider et al.. 20.1.6; Barkur and Bairv. 20.1.5; Weston et al.. 20.1.4; Corv-
Sleehta et al.. 20.1.3) and a few studies began exposure of the dam at the time of conception (GD 0) (Rao
Barkur and Bairv. 20.1.6; Barkur and Bairv. 20.1.5; Barkur et al.. 20.1.1'). Of note is that Teiion et al. (2006).
a study discussed in the 2013 Pb ISA, elaborated that the observed reduction in litter weights born to Pb-
treated dams was largely driven by the reduced size of female pups, whereas males were unaffected. In
agreement, some recent studies that reported no effect of Pb on birth weight assessed weight in male pups
only (Barkur and Bairv. 2015; Barkur et al.. 20.1.1). However, all other recent studies included females in
birth weight analyses and reported no effects of Pb on birth weight of exposed offspring.

8.3.2.3	Integrated Summary of Effects on Prenatal Growth

The epidemiologic studies in the 2013 Pb ISA reported associations between maternal bone Pb
and low birth weight and with studies of Pb air exposures and birth weight. The associations were less
consistent when using maternal blood Pb or umbilical cord and placenta Pb as the exposure measurement
although some studies did demonstrate associations. The studies of Pb exposure and fetal growth were
limited by cross-sectional study design, small sample size, high Pb concentrations (air Pb as high as
30 |ig/m3). and in some studies, the lack of control of confounders. A recent quasi-experimental study of
maternal exposure to airborne Pb during pregnancy found an increase in birth weight, decreased
probability of low birth weight, and reduction in the probability of SGA after the deleading of racing fuel.
However, overall, the recent epidemiologic studies reported inconsistent associations between Pb

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exposure and prenatal growth outcomes, while the toxicological studies consistently reported no effects of
Pb on offspring birth weight. The inconsistent findings from the recent epidemiologic studies may be due
to differences in study design, timing of when the exposure was measured (e.g., during pregnancy, at
delivery), biomarkers examined for Pb (e.g., maternal blood, cord blood, maternal red blood cells,
maternal serum, placental tissue), difference in growth metrics assessed (e.g., birth weight, birth length,
GA), and small sample sizes in some studies. While there were inconsistencies in the findings among the
epidemiologic studies, the recent epidemiologic studies were able to address a few of the uncertainties in
the 2013 Pb ISA. Many of the recent studies were conducted in well-designed longitudinal birth cohorts,
considered the differences in effects by infant sex, and controlled for wide range of confounders,
including GA (when not an outcome of interest), and maternal health factors (e.g., smoking, parity, BMI).
Additionally, some epidemiologic studies controlled for other metal exposure, and other studies evaluated
the associations with joint effects or as a mixture. A few toxicological studies were reviewed in the 2013
Pb ISA, all of which reported reductions in birth weight of offspring born from Pb-exposed dams.
However, recent toxicological studies do not support previous studies and consistently report no effects of
Pb on offspring birth weight.

8.3.3 Preterm Birth

The recent epidemiologic and toxicological studies that examined the relationship between Pb
exposure and preterm birth are summarized in the text below. Study details of the recent epidemiologic
studies are included in Table 8-5 and the recent toxicological studies are in Table 8-3.

8.3.3.1 Epidemiologic Studies on Preterm Birth

The epidemiologic studies reviewed in the 2013 Pb ISA reported overall inconsistent findings
regarding a relationship between indicators of Pb exposure and preterm birth. However, there were a few
well-conducted epidemiologic studies that reported associations between maternal blood Pb and preterm
birth (V igeh et ah. 20.1.1; Jelliffe-Pawlowski et ah. 2006).Among the epidemiologic studies , there were
no apparent patterns within the type of exposure measurement or Pb level. Many of these studies are
limited by the small number of preterm births and their cross-sectional design (i.e., studies of umbilical
cord blood may not adequately characterize BLLs earlier in pregnancy). Among the longitudinal cohort
studies, the results were mixed, with some studies reporting associations between maternal blood Pb
during pregnancy and preterm birth. Most studies controlled for potentially important confounders, such
as maternal age and smoking.

In the recent epidemiologic studies examining the risk of preterm birth and Pb exposure, the
findings were generally consistent (Table 8-5). Most notably is a quasi-experimental study employing
difference-in-difference methodology. In a study by Bui et ah (2022). the effects of short-term maternal

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exposure to airborne Pb during pregnancy on preterm birth was estimated using a quasi-experimental
variation in airborne Pb exposure based on NASCAR's deleading of racing fuel in a difference-in-
difference model in the CCG MSA in North Carolina. There was decreased probability of preterm birth of
0.295 (95% CI: -0.0572, -0.000185) among children born to mothers residing less than 4000 meters of
the CMS, compared to those residing greater than 10,000 meters after deleading of racing fuel. The
difference-in-difference methodology allows for the control of time-varying confounders, removing
biases from comparisons over time in the treatment group that could be the result of trends due to other
causes of the outcome.

In a cross-sectional study of 696 mother-infant pairs in the Study on the Environment and
Reproductive Health (EMASAR) cohort in Argentina, the relationship between maternal Pb levels, which
were collected 36 ± 12 hours postpartum, and preterm birth was examined ("Xu et ah. 2022b). Among
tertiles of maternal Pb levels, there were null associations with preterm birth (OR: 1.24 [95% CI: 0.35,
4.4] in tertile 2 and OR: 1.26 [95% CI: 0.32, 5.00] interfile 3). In another study, cord blood samples were
obtained from 432 infants born in an area with e-recycling (Guiyu) and 99 from an area without e-
recycling (Xiamen) in China, but there was no increased risk of preterm birth (OR: 1.09 [95% CI: 0.93,
1.28]) ("Xu et al.. 2012). Additionally, another study used placental tissue Pb levels in association with
preterm birth among 327 mother-infant pairs who were part of the Instituto de Nanociencia y Materiales
de Aragon (INMA) Project in Spain and found no association with risk of preterm birth (OR: 0.40 [95%
CI: 0.04, 4.70]) (Freire etaL 2019).

In a case-control study, maternal serum Pb, collected during the first or second trimester, was not
associated with risk of spontaneous preterm birth (OR: 1.46 [95% CI: 0.97, 2.18]) among 147 cases and
381 controls (Yu et al.. 20.1.9'). When stratified by the trimester of collection of maternal serum Pb. there
was null association for spontaneous preterm birth (OR: 1.63 [95% CI: 0.91, 2.91]) with first trimester
maternal serum Pb only or second trimester maternal serum Pb only (OR: 1.27 [95% CI: 0.71, 2.28]). In a
nested case-control study, the association between exposure to 41 metals/metalloids, including Pb, during
early pregnancy measured in maternal serum and risk of spontaneous preterm birth was investigated (Xu
et al.. 2022a). There were 74 cases of spontaneous preterm birth and 74 controls. In the highest quartile of
maternal serum Pb levels, there was an increased risk of spontaneous preterm birth of 4.09 (95% CI: 1.31,
12.77) and there was evidence of potential exposure-response across the quartiles (p for trend: 0.017).

Tsuii etal. (20.1.8) used data on 14,847 pregnant women who were participants of the JECS to
assess the association between second and third trimester maternal blood (collected at gestational weeks
14-39) and early preterm (22 to <34 weeks) and late preterm (34 to <37 weeks). Among the quartiles of
Pb exposure, there was no increased risk in early preterm birth or late preterm birth. There was also no
evidence of a linear exposure-response trend among the Pb exposure quartiles in either the early preterm
births (p for trend: 0.134) or late preterm (p for trend: 0.920). In another cohort study using data from the
JECS, per each 0.1 (ig/dL increase in maternal BLL, there was no increased risk of preterm delivery (OR:
0.90 [95% CI: 0.70, 1.16]) (Goto et al.. 2021).

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In a small cohort (n = 98) from the Conditions Affecting Neurocognitive Development and
Learning in Early Childhood (CANDLE) study in Shelby County, TN, Pb was measured cord blood and
from maternal blood collected during the second and third trimester, at delivery ("Rabito et ah. 20.1.4').

Each 0.1-unit increase in maternal blood Pb in the second trimester (OR: 1.66 [95%CI: 1.23, 2.23]) and
third trimester (OR: 1.24 [95% CI: 1.01, 1.52]) was positively associated with preterm birth, but there was
no increased risk of early-term birth (>37 to <39 weeks) associated with maternal blood Pb in the second
trimester (OR: 0.87 (95% CI: 0.63, 1.20]) and third trimester (OR: 0.88 [95% CI: 0.69, 1.13]).

In the Avon Longitudinal Study of Parents and Children, maternal blood samples were collected
as early as possible in pregnancy, with a median GA of 11 weeks at the time of sampling (range 1-
42 weeks, IQR 9-13 weeks) ( l as 1 or et ah. 2015). There was increased risk of preterm delivery (OR: 2.00
[95% CI: 1.35,3.00]) for maternal BLLs >5 (.ig/dL. Li et ah (20.1.7a) investigated the associations between
maternal serum Pb levels and risk of preterm birth in a population-based birth cohort (n = 3,125), part of
the China-Anhui Birth Cohort. Maternal serum Pb levels were categorized into tertiles: low-Pb
(<1.18 (ig/dL), medium-Pb (1.18-1.70 (.ig/dL). and high-Pb (>1.71 (.ig/dL). There was an increased risk of
preterm birth in the medium-Pb tertile (OR: 2.33 [95% CI: 1.49, 3.65]) and high-Pb tertile (OR: 3.09
[95% CI: 2.01,4.76]).

In a study by Ash rap et ah (2020). individual and mixture effects of metals and metalloids on
preterm birth among 731 pregnant women in the PROTECT cohort were examined. Maternal blood was
collected at 16-20 and 24-28 weeks gestation. There was an increased risk of preterm birth (OR: 1.63
[95% CI: 1.17,2.28]) and spontaneous preterm birth (OR: 1.53 [95% CI: 1.00, 2.35]) per IQR increase in
maternal blood Pb in the individual pollutant model. The mixture pollutant models and elastic net
regularization identified Pb and zinc as the most important predictors of preterm birth, while BKMR
method identified Pb, zinc, and Mn as most predictive of preterm birth. In another study in the PROTECT
cohort, the modifying effect of psychosocial stress on the association between Pb and overall preterm
birth (<37 completed weeks of gestation) and spontaneous preterm birth (<37 completed weeks of
gestation defined as presentation of premature rupture of the membranes, spontaneous preterm labor, or
both) (Ashrap et ah. 2021). There was an increased risk of overall preterm birth among mothers who
reported "good" psychosocial status (OR: 1.72 [95% CI: 1.14, 2.58]), but null association among mothers
who reported "poor" psychosocial status (OR: 1.43 [95% CI: 0.69, 2.97]). There were null associations
among mothers who reported "good" psychosocial status and "poor" psychosocial status and spontaneous
preterm birth (OR: 1.56 [95% CI: 0.93, 2.6] and OR: 1.22 [95% CI: 0.42,3.56], respectively).

8.3.3.2 Toxicological Studies on Preterm Birth

Both the 2013 Pb ISA and the 2006 Pb AQCD did not describe any studies that reported on the
effects of Pb on preterm birth in animals. Only one recent study was found that reports on gestation
duration (Betharia and Maher. 20.1.2). Betharia and Maher (20.1.2) reported no effect of Pb on gestation

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term when Sprague-Dawley rats were dosed from GD 0 to postnatal day (PND) 20. BLLs were measured
in offspring and reported to be 9.03 (ig/dL on PND 2, 0.976 (ig/dL on PND 25, 0.0318 (ig/dL on PND 60.

8.3.3.3 Integrated Summary of Effects on Preterm Birth

In summary, there were inconsistencies in the recent epidemiologic studies examining the
relationship between Pb exposure and risk of preterm birth, similar to the 2013 Pb ISA. There was no
apparent pattern associated with any biomarker of Pb exposure. Several of the recent epidemiologic
studies were conducted in well-designed, longitudinal birth cohorts, and controlled for wide range of
confounders, including GA, other metals, and maternal health factors (e.g., smoking, parity, BMI).
Overall, among the epidemiologic studies, there was a pattern of elevated risk of preterm birth observed
across several studies from multiple geographic locations, including a quasi-experimental study. Among
these studies, there were still some uncertainties in the timing of the exposure (e.g., during pregnancy, at
delivery), and biomarkers examined for Pb (e.g., maternal blood, cord blood, maternal red blood cells,
maternal serum, placental tissue). There were no toxicological studies that investigated the effects of Pb
on preterm birth in the 2013 Pb ISA and the 2006 Pb AQCD, and recent toxicological data are sparse with
only a single PECOS-relevant study available, which reported no effects of Pb on gestation duration,
making it difficult for toxicological data to support epidemiologic evidence.

8.3.4 Birth Defects

The recent epidemiologic and toxicological studies that examined the relationship between Pb
exposure and birth defects are summarized in the text below. Study details of the recent epidemiologic
studies are included in Table 8-6 and the recent toxicological studies are in Table 8-3.

8.3.4.1 Epidemiologic Studies on Birth Defects

In the 2013 Pb ISA, there were only a few studies available for review evaluating associations
between Pb exposure and birth defects, specifically NTDs. These studies did not report associations
between Pb exposure and NTDs. These studies were limited by the timing of Pb measurements, whether
taken at delivery or postnatally, and the lack of potential confounders.

A few recent epidemiologic studies examined the relationship between Pb levels and birth
defects. Several studies evaluated the association between NTDs in different biomarkers (placental tissue,
umbilical cord tissue, and maternal serum) (Liu et al.. 2021; Tian et ah. 2021; Jin et ah. 2013). Recent
studies that measured Pb exposure from placental tissue or umbilical cord tissue reported no increased
risk for NTDs overall or by subtype (Liu et ah. 2 ; et ah. 2013) (see Table 8-6 for details).

However, in a case-control study which evaluated the single and joint effects of 10 metals measured in

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maternal serum during pregnancy, there was increased risk for NTDs ("Tian et ah. 202 D. In the single
pollutant model, there was increased risk of NTD of 2.05 (95% CI: 1.05, 4.02) in the second tertile and
3.51 (95% CI: 1.76, 6.98) in the third tertile, relative to the lowest tertile of maternal serum Pb levels,
indicating an exposure-response relationship (p for trend: <0.001). There was also increased risk by NTD
subtype. There was increased risk of spina bifida of 2.16 (95% CI: 1.00, 4.88) in the second tertile and
5.16 (95% CI: 2.24, 11.87) in the third tertile, relative to the lowest tertile of maternal serum Pb levels,
with an indication of an exposure-response relationship (p for trend: 0.022). For anencephaly, there was
increased risk of 2.97 (95% CI: 1.09, 8.12) in the second tertile and 5.54 (95% CI: 1.89, 16.19) in the
third tertile, relative to the lowest tertile of maternal serum Pb levels, with an indication of an exposure-
response (p for trend: 0.002). Among female infants, there was increased risk of NTD of 6.45 (95% CI:
2.20, 18.95) in the highest tertile, relative to the lowest tertile of maternal serum Pb levels, with exposure-
response relationship (p for trend: 0.001). Among male infants, there was increased risk of NTD of 2.16
(95% CI: 1.03, 4.59), and an indication of an exposure-response relationship (p for trend: 0.048).

Pi etal. (20.1.8) investigated the associations between placental Pb concentrations and the risk of
orofacial cleft (OFC) defects among 103 cases and 206 controls in northern China. With increasing
tertiles of placenta Pb concentrations (p for trend <0.001), there was increased odds of orofacial defects of
3.88 (95% CI: 1.78, 8.42) for those in the second (57.5-96.8 ng/g dry weight) tertile of placenta Pb
exposure and 5.17 (95% CI: 2.37, 11.29) for those in the highest (>96.8 ng/g dry weight) tertile of
placenta Pb exposure, compared to the lowest (<57.5 ng/g dry weight). When restricting to those with
higher than the median placenta Pb concentration (>77.2 ng/g), there was increased risk of 3.08 (95% CI:
1.74, 5.47) of OFC defects among 71 cases and 84 controls. However, in a nested case-control study
among a subset of participants in the JECS, Takeuchi et al. (2022) did not find increased risk of cleft lip
and palate (n = 192 cases and n = 1,920 matched controls) and second trimester maternal blood Pb
concentrations (OR: 1.10 [95% CI: 0.55, 2.21]), which controlled for co-exposure to three other metals
(Hg, Cd, and Mn) in the multivariate model.

In another study of the JECS, maternal serum Pb samples were collected during mid- and late
gestation and were evaluated in association with congenital abdominal malformations (Mivashita et al..
202.D. There were 139 cases and 89,134 controls. There were null associations across the quartiles of
maternal serum Pb concentrations and any abdominal malformations, with no exposure-response
relationship across quartiles (p for trend: 0.233). The null associations persisted for the subtypes of
congenital abdominal malformations, but there was an inverse exposure-response relationship observed
across the quartiles of maternal serum Pb and omphalocele (p for trend: 0.033).

A single study explored the associations between umbilical cord serum Pb levels and congenital
heart disease (CHD) birth defects among 97 case and 201 controls (Liu et al.. 20.1.8). In the highest
umbilical serum Pb group (>8.26 ng/mL), the odds of CHD were 1.67 (95% CI: 0.88, 3.17) compared to
the those in the lowest umbilical serum Pb group (<6.69 ng/mL). The odds by CHD subtypes were near
null, (CIs include 1) (see Table 8-6).

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8.3.4.2

Toxicological Studies on Birth Defects

The 2013 Pb ISA did not report any toxicological studies that investigated the effects of Pb on
birth defects. The 2006 Pb AQCD described studies that reported Pb-induced birth defects; however,
these findings were confounded by maternal toxicity (Dev et ah. 2001; Ron is et ah. .1.996; Flora and
Tandon. 1987). Two recent studies published since the 2013 Pb ISA have investigated Pb-induced birth
defects in offspring in rodents (Table 8-3). Both studies dosed Wistar rats with 0.2% Pb in the drinking
water for varying duration, including dosing starting 30 days prior to gestation and ending the day prior to
mating, dosing from GD 0 to PND 21, and dosing from GD 0 to 21 (Rao Barkur and Bairv. 20.1.6; Barkur
and Bairv. 2015). Offspring BLLs measured on PND 22 varied between 3.02-3.03 (.ig/dL for animals
from dams dosed prior to gestation, 5.30-5.51 (ig/dL for animals from dams dosed during gestation, and
31.6-32.0 (ig/dL for animals from dams dosed from the beginning of gestation to lactation day 21. No
maternal toxicity was apparent in any of the recent studies, suggesting that the contrast found between the
lack of malformations observed in these recent publications and the reported malformations described in
the 2006 Pb AQCD may be attributed to a lack of maternal toxicity due to the use of lower doses in more
recent studies.

8.3.4.3 Integrated Summary of Effects on Birth Defects

The studies reviewed in the 2013 Pb ISA did not report associations between Pb exposure and
NTDs. Among the recent epidemiologic studies, there were inconsistent associations with Pb exposure
and NTDs, congenital heart defects, and OFC defects overall. While the associations were generally null
for NTDs and CHDs, there was a pattern of positive associations with OFC defects. The inconsistencies
in these findings are limited by the different birth defects of interest, the small sample sizes given the rare
outcome, timing of Pb exposure (different measurements to estimate exposure during pregnancy),
differences in the biomarker tested, and the confounders considered in the analyses. The recent
epidemiologic studies controlled for a wide range of potential confounders; however, which was a
limitation from the 2013 Pb ISA. Further, some studies considered co-exposure to other metals and
differences by infant sex. Some previous toxicological studies reported that Pb exposure resulted in birth
defects in offspring, but it was noted that these studies often used doses so high that maternal toxicity
occurred as well. Recent toxicological studies report no effects of Pb on birth defects in offspring and also
do not report that maternal toxicity occurred, further supporting that maternal toxicity may have been
involved with the birth defects observed in previous studies.

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8.3.5 Spontaneous Abortion and Pregnancy Loss and Fetal and Infant
Mortality

The 2013 Pb ISA concluded that the toxicological and epidemiologic data provided inconsistent
findings for the role of Pb in spontaneous abortions, while there were no available epidemiologic or
toxicological studies on the relationship between Pb levels and infant mortality. The recent epidemiologic
and toxicological studies examining the relationship between Pb exposure and spontaneous abortion,
pregnancy loss, and fetal and infant mortality are summarized in the text below. Study details of the
recent epidemiologic studies are included in Table 8-7 and the recent toxicological studies are in
Table 8-3.

8.3.5.1 Epidemiologic Studies on Spontaneous Abortion and Pregnancy Loss and
Fetal and Infant Mortality

In the 2013 Pb ISA, there was a limited number of epidemiologic studies that examined Pb
exposure and spontaneous abortion or pregnancy loss with inconsistent findings. Studies that examine
spontaneous abortion or pregnancy loss are difficult to conduct, as many spontaneous abortions or
pregnancy losses occur during the first trimester. Women may miscarry before being enrolled in a study
and/or women may not have known they were pregnant when they miscarried, further limiting the ability
to detect subtle effects, especially if higher Pb exposures do lead to increased risk of early spontaneous
abortions or pregnancy loss. In addition, some studies are limited by their retrospective examination of
current Pb biomarker levels in relation to previous miscarriages. The epidemiologic studies reviewed in
the 2013 Pb ISA had limited sample sizes and little control for potential confounding factors, with some
studies including no potential confounders in their analyses. There were no epidemiologic studies of Pb
exposure and fetal and infant mortality reviewed in the 2013 Pb ISA and there were no recent PECOS-
relevant epidemiologic studies of Pb exposure and fetal and infant mortality.

There were only a few recent epidemiologic studies that evaluated Pb exposure and spontaneous
abortion and pregnancy loss. There were inconsistent findings among the studies and no apparent pattern
of association by biomarker of Pb exposure. In a recent study, cord blood samples were obtained from
432 infants born in an area with e-recycling (Guiyu) and 99 from an area without e-recycling (Xiamen) in
China ("Xu et ah. 2012). There was an increased risk of 4.20 (95% CI: 3.40. 5.18) of stillbirth rate with
cord BLLs comparing infants from the area with e-recycling (Guiyu) compared to infants from the area
without e-recycling (Xiamen). In a recent cohort study, couples (n = 344) were prospectively followed to
explore the relationship between blood Pb concentrations at enrollment and with pregnancy followed to
estimate the risk of incident of pregnancy loss (Louis et ah. 20.1.7). Each participant's blood Pb
concentration and time to pregnancy loss was modeled individually and as a couple. In the individual
partner models, there was no increased risk of pregnancy loss for female partner blood Pb (hazard ratio
[HR]: 1.01 [95% CI: 0.82, 1.25]) or male partner blood Pb (HR: 0.95 [95% CI: 0.77, 1.17]). In the
couple-based model, the associations were unchanged (female partner HR: 1.01 [95% CI: 0.80, 1.28] and

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male partner HR: 0.96 [95% CI: 0.77, 1.22]). Among a cohort of 166 women in Iran, there was no
increased risk (OR: 1.08 [95% CI: 0.98, 1.20]) of spontaneous abortion with maternal BLLs in early
pregnancy (Vigeh et ah. 2021). In another prospective cohort among women seeking treatment at a
fertility clinic in Turkey, blood Pb concentrations were assessed in association with ongoing pregnancy
(Tolunav et ah. 20.1.6'). The study participants were categorized into patients with ongoing pregnancy
(n = 20) and patients who experienced assisted reproductive technology (ART) failure, miscarriage, or
biochemical pregnancy (n = 81). There was a 2.2% lower risk (relative risk [RR]: 0.978 [95% CI: 0.957,
0.999]) for ongoing pregnancy for each 1 (ig/dL higher blood Pb concentration. Among a cohort of 1,184
women undergoing assisted reproductive therapy in China, associations between maternal serum Pb
concentrations and spontaneous abortion before gestational week 12 were evaluated (Li et ah. 2022).
There was an increased risk of 1.39 (95% CI: 1.02, 1.91) of spontaneous abortion before gestational week
12 with increasing maternal Pb serum levels. When categorized into tertiles, the associations between
maternal Pb serum levels and spontaneous abortion before gestational week 12 were null.

8.3.5.2 Toxicological Studies on Spontaneous Abortion and Pregnancy Loss and
Fetal and Infant Mortality

The 2013 Pb ISA did not report any toxicological studies that investigated the effects of Pb on
offspring mortality at any stage of development. Some studies that investigated the effects of Pb on
offspring mortality were summarized in the 2006 Pb AQCD. Overall, these studies found that gestational
exposure increased pregnancy loss and implantation loss (BLLs >32 (.ig/dL) (Pinon-Lataillade et ah.
.1.995; Singh et ah. .1.993; Piasek and Kostial. 1.991; Logdberg et ah. 1987). Some recent studies have also
investigated the effects of Pb exposure on offspring mortality (Table 8-3). However, recent studies
reported that Pb did not have effects on measures of pre- or postnatal mortality, including litter size.
Rodent studies that dosed prior to and during gestation reported no increase in stillbirth or decrease in
number of pups born to treated dams ("Saleh et ah. 20.1.8; Rao Barkur and Bairv. 20.1.6; Barkur and Bairv.
20.1.5; Weston et ah. 20.1.4; Corv-Slechta et ah. 2( . liaria and Maher. 20.1.2). BLLs, sources
(e.g., BLLs from dams or BLLs from offspring), and times of measurement were variable between these
studies (0.0318-27.7 (j,g/dL; GD 20-PND 60), but in general, BLLs in recent studies were lower than
those reported in previous studies. The contrast in the effects of Pb exposure on offspring mortality
observed between previous studies and recent studies may be attributed to the lower BLLs achieved in
recent studies compared to the higher BLLs in previous studies.

Postnatal offspring mortality was also investigated in some rodent studies, and some studies
reported on measures of offspring mortality that included postnatal death and survival until certain
timepoints after birth (e.g., weaning). These studies also did not report any effects of Pb exposure on
postnatal survival. Most studies utilized dosing paradigms that dosed before or during gestation (Barkur
and Bairv. 20.1.5; Betharia and Maher. 20.1.2) and reported BLLs at different times postnatally (PND 2-60;
0.0318 (ig/dL-5.30 (ig/dL) with BLLs tending to be lower in time points with the longest amount of time

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since cessation of exposure. Some studies utilized a dosing paradigm that exclusively exposed animals
postnatallv ("Barkur and Bairv. 21 . tharn et aL 20.1.1). In agreement, these studies also reported no
effects of offspring mortality during postnatal time points (PND 4-29; BLLs 3.27-26.65 (.ig/dL).

8.3.5.3 Integrated Summary of Effects on Spontaneous Abortion and Pregnancy Loss
and Fetal and Infant Mortality

The 2013 Pb ISA reported inconsistent findings from the epidemiologic studies on Pb exposure
and spontaneous abortion and pregnancy loss. The findings from recent epidemiologic studies on Pb
exposure and spontaneous abortion and pregnancy loss were also inconsistent. A single study reported
increased risk of stillbirth with cord blood Pb. While recent cohort studies among healthy participants did
not find increased risk of pregnancy loss or spontaneous abortion, women seeking treatment from a
fertility clinic had increased risk of spontaneous abortion before gestational week 12 or decreased risk of
an on-going pregnancy. The women seeking fertility treatment that were recruited as participants may be
different from those in the general population, limiting the generalizability of the results as the study
populations may not be representative of the general population as they have already been diagnosed and
are seeking treatment for fertility issues. However, early pregnancy loss is more likely to be ascertained
from women seeking treatment at fertility clinics. Previous toxicological studies reported increased rates
of pregnancy loss and implantation loss in animals dosed with Pb during gestation. This contrasts with
more recent literature which did not report any effect of Pb on pre- or postnatal offspring mortality.
Although not always consistent, BLLs were generally lower in recent toxicological literature when
compared to previous literature, possibly explaining the observed contrast in results.

8.3.6 Placental Function

In the 2013 Pb ISA, there were no epidemiologic or toxicological studies available that evaluated
Pb concentrations and associations with placenta function. Recent epidemiologic and toxicological studies
evaluating the association between Pb exposure and placental function are limited. The epidemiologic
studies were cross-sectional studies. Study details for the recent epidemiologic studies are included in
Table 8-8 and the toxicological studies are included in Table 8-3.

8.3.6.1 Epidemiologic Studies on Placental Function

In the 2013 Pb ISA, there were no epidemiologic studies available that evaluated Pb
concentrations and associations with placenta function. In recent cross-sectional epidemiologic studies,
there were different markers of placental function evaluated. One marker of placental function that was
evaluated was placental thickness, which can restrict intrauterine fetal growth ("Al-Saleh et aL 20.1.4).
Maternal BLLs measured at delivery were found to be associated with the risk of placental thickness

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below the 10th percentile (OR: 1.64 [95% CI: 1.12, 2.41]). In another study, using a cross-section from
the JECS, the relationship between maternal blood Pb collected during the second trimester and placental
previa and placenta accreta among 16,019 women was examined (Tsuii et ah. 20.1.9'). Placenta previa is a
condition in which the placenta is attached to the lower uterine segment and completely or partially
covers the internal cervix, and when chorionic villi abnormally invade to myometrium, placenta accreta
occurs ("Tsuii et ah. 20.1.9). There was increased odds of placenta previa in the second quartile (4.80-
5.95 ng/g) of maternal blood Pb (OR: 2.59 [95% CI: 1.40, 4.80]), but null associations the third quartile
(5.96-7.44 ng/g) maternal blood Pb (OR: 1.32 [95% CI: 0.66, 2.64]) and fourth quartile (>7.45 ng/g)
maternal blood Pb (OR: 1.34 [95% CI: 0.67, 2.67]). There were null associations for placenta accreta
across the blood Pb quartiles (Table 8-8).

8.3.6.2	Toxicological Studies on Placental Function

The 2013 Pb ISA reported a single study that investigated the effects of Pb exposure on placental
function. Wang et ah (2009) reported decreased placental weight in Wistar rats along with dose-
dependent increasing pathology of cytoarchitecture and cytoplasmic organelles. Focusing on different
gestational periods, this study exposed dams to 0.025% Pb via drinking water from either GD 1-10,
GD 11-20, or GD 1-20 (maternal BLLs on GD 20 were 26.3, 12.4 (ig/dL, and 36.0 (ig/dL, respectively).
Some recent studies have reported on similar placental outcomes (Table 8-3). Wang et ah (20.1.4) also
dosed using the same dosing paradigm (dosing during GD 1-10, GD 11-20, or GD 1-20 via drinking
water) and reported that placentae collected from pregnant Wistar rats on GD 20 showed similar dose-
dependent decreases in weight and histopathological abnormalities such as vascular congestion,
trophoblast degeneration, chorionic villi interstitial edema, irregularity of trophoblast cells in the labyrinth
and trophospongium, degeneration of trophoblast cells, and chorionic villi vacuolization (maternal Pb
levels on GD 20 were reported to be between 12.4-36.0 (ig/dL and varied by dosing window). Two other
studies that dosed pregnant Sprague-Dawley rats via gavage from GD 0-20 and similarly reported
reduced placental weights (maternal blood Pb on GD 20 was 23.9-27.7 (.ig/dL) (Saleh et ah. 20.1.9; Saleh
et ah. 2018). Of note is that both of these recent studies by Saleh et al. ("Saleh et ah. 20.1.9; Saleh et ah.
20.1.8) also reported reduced brain weights in dams which is indicative of overt toxicity. Thus, it is
possible the altered placental weight could be attributed to overt toxicity experienced by the dams.

8.3.6.3	Integrated Summary of Effects on Placental Function

There were no epidemiologic studies available that evaluated Pb concentrations and associations
with placenta function in the 2013 Pb ISA. The recent epidemiologic studies reviewed that assessed the
relationship of Pb exposure and placental are limited. The differences in the different markers of placental
function make it difficult to judge coherence and consistency across these studies, but these positive
associations are an indication that exposure to Pb may result in effects on placental function during

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pregnancy. Previous toxicological data on the effects of Pb on placental weight are limited to a single
study which reported decreased placental weight and histological alterations. Recent studies also reported
that dams dosed with Pb had reduced placental weight, but of note is that these studies also reported
reduced brain weight in dams, suggesting that overt toxicity may have occurred and could be related to
the observed reductions in placental weight.

8.3.7 Other Pregnancy and Birth Outcomes

There were several recent studies that evaluated associations between Pb exposure and other
pregnancy and birth outcomes in the epidemiologic and toxicological literature. More specific study
details for the epidemiologic studies, including Pb levels, study population characteristics, potential
confounders, and select results from these studies are highlighted in Table 8-9. Specific study details for
the toxicological studies are provided in Table 8-3.

8.3.7.1 Epidemiologic Studies on Other Pregnancy and Birth Outcomes

There were several recent studies with other outcomes related to pregnancy and birth. More
specific study details, including Pb levels, study population characteristics, potential confounders, and
select results from these studies are highlighted in Table 8-9. In studies of other pregnancy and birth
outcomes, maternal Pb blood concentrations were associated with high levels of leptin, a fetal marker of
metabolic function ("Ashley-Martin et ah. 20.1.5a'); and cord blood Pb concentrations were negatively
associated with cord blood relative telomere length (rTL) (Herlin et ah. 20.1.9'). However, there were null
associations in several other studies evaluating Pb exposure and outcomes related to pregnancy and birth.
There was a null association between maternal serum Pb levels and nuchal translucency, which is the
subcutaneous space in the fetal neck and is visible with ultrasound imaging in the first trimester (Liao et
ah. 20.1.5'). Increased nuchal translucency thickness in the first trimester has been reported to be a risk
factor for chromosomal abnormalities, genetic syndromes, congenital heart defects, structural
abnormalities, intrauterine infection, neurodevelopmental delay, and fetal demise ("Liao et ah. 20.1.5').
There was null associations between maternal blood Pb concentrations and thymic stromal lymphopoietin
(TSLP) and interleukin-33 (IL-33), which are biomarkers of fetal immune system (Ashley-Martin et ah.
20.1.5b'); maternal blood Pb concentrations and elevated cord blood concentrations of immunoglobulin E
(IgE) (Ashley-Martin et ah. 20.1.5b'); and maternal blood Pb and markers of fetal metabolic function (low
leptin, low adiponectin, and high adiponectin) (Ashley-Martin et ah. 2015b'). Additionally, there were
inconsistent associations between maternal BLLs during pregnancy and secondary sex ratio. Among
participants in the ALSPAC in the United Kingdom, Taylor et ah (20.1.4) reported no associations among
quintiles of maternal Pb levels during the first trimester of pregnancy and secondary sex ratio (odds of
having a male child). Among the participants of the Longitudinal Investigation of Fertility and the
Environment (LIFE) cohort, there were associations with secondary sex ratio (ratio of live male to female

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births, reflecting a male excess) and both maternal and parental blood samples were measured for Pb at
baseline (before pregnancy) (Bloom et ah. 20.1.5). However, Tatsuta et al. (2022b) reported increased odds
of male births (secondary sex ratio) of 1.279 (95% CI: 1.224, 1.336) in the highest quintile of maternal
blood Pb among a subset of participants in the JECS.

8.3.7.2	Toxicological Studies on Other Pregnancy and Birth Outcomes

The 2013 Pb ISA summarized a single study reporting on other birth outcomes such as sex ratio.
Dumitrescu et al. (2008a) reported an increased female:male ratio in offspring born to Wistar rats that
were dosed with 100 or 150 ppb of Pb in the drinking water starting 3 months prior to mating and
continuing until birth. This study did not report on BLLs in dams or offspring, resulting in more
challenges when comparing this study to more recently published studies (Table 8-3). Weston et al.
(20.1.4) was the only recent study to report an effect of Pb exposure on the sex ratio of pups born to
exposed dams. In agreement with Dumitrescu et al. (2008a). Weston et al. (20.1.4) reported that Long-
Evans rats dosed starting 76 days prior to mating and continuing through birth gave birth to female
skewed litters when compared to control. However, it is worth noting that in Weston et al. (20.1.4) control
litters had unusually high numbers of males that resulted in a 1.5 male:female ratio of pups born, whereas
Pb-treated litters had a more even distribution that resulted in a ratio closer to 1:1 (BLLs were 14.6 and
15.7 (ig/dL in female and male pups, respectively, on PNDs 5-6). Other recent studies contrast with these
studies and report no effects on sex ratio in Pb-treated females. One study using a dosing paradigm
similar to the two studies above (exposure beginning 2 months prior to dosing and continuing through
birth) reported no effects of Pb on sex ratio (BLLs 12.12 (ig/dL in dams at weaning) in C57BL/6 mice
(Corv-Slechta et al.. 2013). Similarly. Tartaglione et al. (2020) dosed Wistar rats from 4 weeks prior to
mating through birth and reported no changes in sex ratio (BLLs 25.5 (ig/dL on PND 23 in pups).
Additional studies in rats dosed females from the beginning of pregnancy through birth and also reported
no changes in sex ratio (BLLs 6.68-9.03 (.ig/dL in pups taken at ages PND 2 and PND 28 in (Betharia and
Mafaer. 20.1.2) and (Baranowska-Bosiacka et al.. 20.1.3). respectively). With the only recent study reporting
alterations in sex ratio also reporting unusual sex ratios in the control group, the effects of Pb exposure on
sex ratio is equivocal.

8.3.7.3	Integrated Summary of Effects on Other Pregnancy and Birth Outcomes

There was a small body of recent epidemiologic studies across various other pregnancy and birth
outcomes; however, the small number of studies limits the ability to judge coherence and consistency
across these studies, although the associations reported demonstrate that Pb exposure could result in
physiological responses that contribute to adverse pregnancy and birth outcomes, such as markers of fetal
metabolic function, fetal immune system biomarkers, and rTL. Toxicological evidence regarding other
pregnancy and birth outcomes are equivocal. While the 2013 Pb ISA reported a study that found that Pb

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exposure led to female-skewed litters, a few recent studies reported no effects of Pb on the ratio of male
to female pups born to Pb-exposed dams.

8.4 Effects on Development

The 2013 Pb ISA (U.S. EPA. 20.1.3') concluded that the collective body of evidence integrated
across epidemiologic and toxicological studies, based on the findings of delayed pubertal onset among
males and females, was sufficient to conclude that there is a causal relationship between Pb exposure and
developmental effects. The current epidemiologic and toxicological studies continue to support
associations between Pb exposure and developmental effects, particularly the delayed onset of puberty in
both males and females. This section does not cover associations between Pb exposure and
neurodevelopmental outcomes, which are discussed in detail in Appendix 3 Nervous System Effects. The
recent epidemiologic and toxicological studies of Pb exposure and effects on development are detailed in
the following sections. The developmental endpoints in subsequent sections are based on postnatal
growth, bodyweight, and stature; puberty among females; and puberty among males.

8.4.1 Effects on Postnatal Growth

The recent epidemiologic and toxicological studies that examine the relationship between Pb
exposure and postnatal growth are detailed below. More specific study details for the epidemiologic
studies, including Pb levels, study population characteristics, potential confounders, and select results
from these studies are highlighted in Table 8-10. Specific study details for the toxicological studies are
provided in Table 8-11.

8.4.1.1 Epidemiologic Studies on Postnatal Growth

The 2013 Pb ISA found inconsistent results between Pb exposure and postnatal growth.
Longitudinal epidemiologic studies had inconsistent findings regarding the association between Pb levels
and postnatal growth. There were further inconsistencies in the findings of the cross-sectional studies
evaluated. While multiple cross-sectional studies reported an association between Pb levels and impaired
growth, several other cross-sectional studies did not report associations between Pb and growth. The
inconsistencies across the studies may be due to study design and differences in the timing of exposure to
Pb (e.g., prenatal, at delivery, or postnatal). However, the longitudinal studies were controlled for
multiple potential confounders, such as age and parity.

There were multiple recent epidemiologic studies that evaluated the relationship between Pb
exposure and postnatal growth. Overall, there were negative associations between Pb exposure and

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specific postnatal growth outcomes among the cross-sectional studies. However, among cohort studies,
there were some inconsistencies in the associations of Pb exposure and different postnatal growth
outcomes. These inconsistencies in the cohort studies may be due to differences in the timing of when Pb
exposure was measured, the biomarker of Pb exposure, and the timing of the outcome.

Among the other cross-sectional studies of Pb exposure and postnatal growth, there were
consistent negative associations. In a National Health and Nutrition Examination Survey (NHANES)
(2013-2016) analysis of 6-11-year-old children (n = 1,634), there were negative associations between an
IQR difference in blood Pb concentrations (median: 0.5 (ig/dL) and standing height ((3: -3.116 cm [95%
CI: -5.03, -1.202]), waist circumference (WC; (3: -5.742 cm [95% CI: -8.769, -2.715]), upper arm
length ([3: -1.068 cm [95% CI: -0.625, -0.512]), and BMI ([3: -2.092 kg/m2 [95% CI: -3.227, -0.957])
(Signes-Pastor et ah. 2021). Among male participants, the negative associations persisted with postnatal
growth outcomes. Among female participants, there were negative associations with BMI, WC, and upper
arm length, but null associations with standing height. Similarly, in a study of primary school children
aged 7-11 years in China, there were negative associations with concurrently measured BLLs (median:
2.61 ng/dL) and height ([3: -3.21 cm [95% CI: -4.24, -2.17]), weight ([3: -1.96 kg [95% CI: -3.11,
0.82]), bust circumference ([3: -2.77 cm [95% CI: -3.79, -1.76]), and waistline ([3: -3.65 cm [95% CI:
-4.78, 2.52]); however, there was a null associations with BMI ([3: 0.20 kg/m2 [95% CI: -0.65, 0.25])
(Kuang et ah. 2020).

When standardizing postnatal growth metrics by Z-score, the associations with Pb exposure were
mixed, even with all median BLLs across studies less than 5 (ig/dL (range: 0.663-4.6 (ig/dL). In a cross-
sectional study, among children <6 years of age (n = 1,678) in China, there were negative associations
between children's logio-BLLs and weight for age Z-score (WAZ) ([3: -0.33 [95% CI: -0.56, -0.11]) and
height-for-age Z-score (HAZ) ([3: -0.38 [95% CI: -0.63, -0.14]), but null associations with BMI-for-age
Z-score (BMIZ) ("Zhou et ah. 2020). When the BLLs were grouped by tertiles. the children in the highest
tertile (>5 ^ig/dL) had lower WAZ ([3: -0.42 [95% CI: -0.62, -0.23]), lower HAZ ([3: -0.36 [95% CI:
-0.58, -0.15]), and lower BMIZ ([3: -0.29 [95% CI: -0.50, -0.07]) than those in the lowest tertile
(<2.5 (ig/dL). The patterns of association held when stratified by child's sex (see Table 8-10). Among
children ranging in age from 8 to 23 months in South Korea, BLLs were associated with post-birth weight
gain (WAZ-BWZ, or the difference of the WAZ at the time of the study and BWZs) ([3: -0.238 [-0.391,
-0.085], standard error [SE]: 0.078) and current HC for age Z-scores (HCAZs; [3: -0.213 [-0.366, -0.06],
SE:0.078) ("Choi et ah. 2017). However, among participants in the Canadian MIREC Child Development
Plus Study, there were no associations reported between blood Pb measured at 2 and 5 years of age and
HAZ, WAZ, or BMIZ overall or when stratified by child's sex (Ashley-Martin et ah. 20.1.9').

Multiple cohort studies examined the relationship between Pb exposure at different time periods
(prenatally and/or at different time periods during childhood) with growth metrics, mainly height and
weight. Among the cohort studies that measured Pb in cord blood, there were inconsistent associations.
While a study among children in Krakow, Poland found no associations with change in mean height over

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a 9-year follow-up period (Jedrvchowski et ah. 2015). a study in the Children's Health and Environmental
Chemicals in Korea (CHECK) study, reported positive associations with Z-scores for weight and BMI at
24 months of age (weight (3: 0.717 [95% CI: 0.195, 1.239] and BMI (3: 0.695 [95% CI: 0.077, 1.313],
respectively) (Kim et al.. 20.1.7). However, there were no associations between cord blood Pb and the Z-
scores of the child's weight, height, or BMI at any other time point (see Table 8-10 for details). When
stratified by children's sex, cord blood Pb was positively associated with an increase of birth height ([3:
0.017 [95% CI: 0.003, 0.031]) and a decrease of PI at birth ([3: -0.055 [95% CI: -0.103, -0.006]) in boys,
but not in girls.

Among participants (n = 1,150) in the Mothers' and Children's Environmental Health (MOCEH)
study in South Korea, maternal BLLs at delivery were negatively associated with Z-scores of weight for
age ([3: -0.33 [95% CI: -0.53, -0.13]) and length for age ([3: -0.30 [95% CI -0.53, -0.08]) at 24 months,
meaning that a 1-fxg/dL increase in late pregnancy Pb levels decreased weight and length at 24 months by
0.33 kg and 0.30 cm, respectively ("Hong et al.. 20.1.4). However, there were no associations between
maternal BLLs in early pregnancy (before 20 weeks gestation) and cord blood Pb with weight and length
(Table 8-10).

Several cohort studies conducted in Mexico measured Pb exposure during pregnancy in maternal
blood, cord blood, and maternal bone and various postnatal growth outcomes. Renzetti et al. (20.1.7)
investigated how Pb exposure during pregnancy is associated with children's growth outcomes, including
height, weight, BMI, and percentage body fat, measured between ages 4-6 years old in a Mexico City
pregnancy cohort (PROGRESS). Maternal blood Pb was measured during the second and third trimester
of pregnancy, as well as at delivery. Cord blood was measured at delivery. Bone Pb levels in the tibia and
patella were also assessed in mothers as a long-term biomarker 1 month postpartum. There were negative
associations between maternal third trimester BLLs and height-for-age ([3: -0.10 [95% CI: -0.19, -0.01])
and weight for age ([3: -0.11 [95% CI: -0.22, -0.003]), but there were no associations between any other
marker of Pb exposure (maternal second trimester blood, cord blood Pb, maternal blood at delivery) and
height-for-age, weight for age, BMI, or percentage of body fat (see Table 8-10). In the Early Life
Exposure in Mexico to Environmental Toxicants (ELEMENT) project, Liu et al. (201.9a) assessed Pb
exposure in maternal bone (as a proxy for cumulative fetal exposure) at 1 month postpartum and also in
blood samples from children annually from 1 to 4 years in association with BMIZ, WC, sum of skinfolds,
and body fat percentage in 248 children aged 8-16 years. Maternal patella Pb levels were associated with
lower child BMIZ ([3: -0.02 [95% CI: -0.03, -0.01]), WC ([3: -0.12 cm [95% CI: -0.22, -0.03]), sum of
skinfolds ([3: -0.29 mm [95% CI: -0.50, -0.08]), and body fat percentage ([3: -0.09% [95% CI: -0.17,
-0.01]). However, there were no associations detected from the postnatal exposure period (blood samples
in children). In another study, children born between 1994 and 2005 in Mexico City had Pb exposure
measured in maternal patella Pb concentrations, a marker of prenatal period exposure, and from infant
and childhood measured in blood at birth to 24 months and 30-48 months (Afeiche et al.. 20.1.2). Among
infants with BLL exceeding the median (4.5 (.ig/dL). there was a decrease in height of 0.84 cm (95% CI:
-1.43, -0.26) compared to children with a level below the median. There were no associations between

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prenatal Pb or childhood Pb and height and there were no associations with BMI at any time point
(prenatal, infancy, or childhood). In cohort of Mexican children aged 6-8 years old, growth (height, HAZ,
and knee height) were assessed in association with BLLs at baseline, after 6 months, and 12 months (Kerr
et al.. 20.1.9). Additionally, as BLLs may differ by the aminolevulinic acid dehydratase (ALAD) genotype,
the authors compared children with the ALAD 1-2/2-2 genotype to children with the ALAD1-1 genotype.
There were negative associations with height ((3: -0.11 cm [95% CI: -0.18, -0.04]), knee height ((3:
-0.04 cm [95% CI: -0.07, -0.02]), and HAZ ([3: -0.02 cm [95% CI: -0.03, -0.01]). Children with
ALAD1-1 had decreased height, knee height, and HAZ, while children with the ALAD1-2/2-2 had reduced
knee height and HAZ, but not height. There were no associations between BLLs and growth at 6- or 12-
month follow-up reported, irrespective of ALAD genotype. This epigenetic study proposes a potential
mechanistic pathway of BLLs differing by genotypes and the associations with growth metrics during
child developmental periods.

There were several studies that explored the associations between Pb exposure and postnatal
growth specifically by sex. In a study by Bums et al. (20.1.7). associations of BLLs were assessed with
longitudinal age-adjusted height (HAZ) and BMI (BMIZ) among male participants in the Russian
Children's Study. Over 10 years of follow-up, after covariate adjustment, boys with higher (>5 (ig/dL)
BLLs compared with lower BLLs were shorter (adjusted mean difference in HAZ: -0.43 [95% CI -0.60,
-0.25]), translating to a 2.5 cm lower height at age 18 years. The decrement in height for boys with higher
BLLs was most pronounced at 12 to 15 years of age (interaction p: 0.03). However, boys with higher
BLLs were leaner (adjusted mean difference in BMIZ: -0.22 [95% CI: -0.45, 0.01]). Deierlein et al.
(20.1.9) used data from the Breast Cancer and the Environment Research Program to investigate
associations of childhood blood Pb concentrations and anthropometric measurements among a multi-site,
multiethnic cohort of girls (n = 683). Blood Pb concentrations were collected before 10 years of age and
height, BMI, WC, percent body fat was measured between 7-14 years of age. There were decreases in
height (range: -2.0 to -1.5 cm), BMI (range: -0.9 to -0.7 kg/m2), WC (range: -3.0 to -2.2 cm), percent
body fat (range: -2.9 to -1.7%) among girls ages 7 through 14 with BLLs of >1 (ig/dL compared to
<1 |ig/dL (Table 8-10).

There were a limited number of studies that examined stunting with exposure to Pb in children. A
single cross-sectional study in a subset of participants in the Interactions of Malnutrition and Enteric
Infections: Consequences for Child Health and Development (MAL-ED) study in Bangladesh reported
increased odds of stunting (OR: 1.78 [95% CI: 1.07, 2.99]) and being underweight (OR: 1.63 [95% CI:
1.02, 2.611) with elevated (>5 (.ig/dL) BLLs. but not wasting (OR: 1.18 [95% CI: 0.64. 2.19]) (Raihan et
al.. 2018). In a cohort study among rural Bangladeshi children. Pb exposure was assessed from umbilical
cord blood at birth and blood Pb at 20-40 months of age with stunting (Gleason et al.. 20.1.6). The odds of
stunting at 20-40 months was 1.12 (95% CI: 1.02, 1.22) per each 1 (ig/dL increase in childhood BLL;
however, there was no association was found between cord BLL and risk of stunting (OR: 0.97 [95% CI:
0.94-1.00]).

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8.4.1.2

Toxicological Studies on Postnatal Growth

The 2013 Pb ISA summarized several current studies and several from the 2006 Pb AQCD that
reported on the effects of Pb on offspring bodyweight and size. The reported effects were fairly
consistent, and nearly all studies reported reductions in bodyweight of offspring exposed to Pb during
developmental periods. Gestational exposure to Pb proved to be sufficient to reduce offspring bodyweight
and body length in rodent studies (Masso-Gonzalez and Antonio-Garcia. 2009; Wang et aL 2009; Teiion
et ah. 2006; Ron is et aL 2001; Ron is et aL 1998a; Ron is et aL 1.998c; Ron is et aL 1.996V The only study
in the 2013 Pb ISA that did not report a reduction in bodyweight was Leasure et al. (2008). which
reported that bodyweight increased at 1 year of age in male C57BL/6 offspring that were dosed starting
prior to conception and ending on PND 10.

Many recent publications have also reported on the effects of Pb-induced changes in bodyweight
of offspring (Table 8-11). In contrast with the 2013 Pb ISA, only some studies reported that Pb exposure
affected offspring bodyweight. One study that dosed Sprague-Dawley rat pups with 1 or 10 mg/kg/d
directly via gavage from PND 4 to 28 reported that bodyweight was decreased in male offspring in both
groups on PND 26 (BLLs 3.27-12.5 (.ig/dL on PND 29) ("Graham et al.. 20.1.1). Another study dosed
Wistar rat dams via drinking water (30 mg/L Pb) from birth until weaning, at which point offspring were
weaned onto the same dosage of Pb in their drinking water as their dam until outcome assessment (de
Figueiredo et al.. 2014). In agreement with Graham et al. (20.1.1). this study by de Figueiredo et al. (2014)
reported reductions in bodyweight of male Wistar rat offspring on PND 60 that were exposed from
conception through PND 60, although female offspring were not evaluated (BLLs 7.2 (ig/dL on PND 60).
Similarly, one study exposed CD-I mice offspring to Pb via dam drinking water (27 or 109 ppm Pb) from
PND 1 to 21 and reported reductions in body weight of pups on PNDs 11, 15, and 19 (BLLs 19.57—
29.16 (.ig /dL on PND 18) (Duan etah. 2017). In contrast, one study conducted in Sprague-Dawley rats
that were exposed from GD 0 to PND 21 via the dam's drinking water (10 |ig/m L Pb) reported increased
bodyweight in offspring on PND 1 and increased bodyweights in females only on PND 49 and 56 (BLLs
9.03 (ig/dL on PND 2, 0.976 (.ig/dL on PND 25, 0.0318 (.ig/dL on PND 60 in pups) (Betharia and Maher.
2012).

Contrasting these recent studies and previous studies discussed in the 2013 Pb ISA are several
studies that reported no effects of Pb exposure on bodyweight in offspring. Studies in both mice and rats
utilizing dosing paradigms that begin exposure prior to conception (Albores-Garcia et al.. 2021; Zhao et
al.. 2021; Sobolewski et al.. 2020; Rao Barkur and Bairv. 20.1.6; Barkur and Bairv. 2015) reported no
effects of Pb exposure on bodyweight at any time point measured (BLLs ranged between 0.4-15.7 (ig/dL
across various time points and studies). Similar null findings were reported in other rodent studies
utilizing other exposure windows including gestational (Rao Barkur and Bairv. 20.1.6; Barkur and Bairv.
20.1.5). lactation (Rao Barkur and Bairv. 20.1.6; Barkur and Bairv. 2015; Basgen and Sobin. 2014). and a
combination thereof (Basha and Reddv. 2015; Barkur et al.. 20.1.1) (BLLs ranged between 2.74-
26.86 Mg/dL).

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8.4.1.3 Integrated Summary of Effects on Postnatal Growth

Overall, among the recent epidemiologic studies, there were negative associations between Pb
exposure and specific postnatal growth outcomes among the cross-sectional studies. However, among
cohort studies, there were some inconsistencies in the associations of Pb exposure and different postnatal
growth outcomes. These inconsistencies in the cohort studies may be the result of the differences in the
timing of Pb exposure measurement (prenatally or postnatally) and the biomarker to measure Pb exposure
(maternal blood, maternal bone, cord blood). Additionally, there was limited evidence that there are
potential differences in the associations between Pb exposure and growth metrics between males and
females. There is also limited evidence of potential epigenetic effects of BLLs differing by genotypes and
the associations with growth metrics during child developmental periods. While cross-sectional studies
are limited by the concurrent measurement of Pb and postnatal growth outcomes, there were several well-
designed cohort studies that support the associations of Pb exposure and decreased growth. These studies
accounted for a wide range of potential confounders, including co-exposure to other metals; however,
some studies did not consider prenatal growth (birth weight, birth length) or maternal characteristics
(height, weight, BMI, smoking), which could potentially influence postnatal growth. While there was a
small body of literature examining the associations between stunting and exposure to Pb, there was
consistent increased odds in stunting with Pb exposure. Previous toxicological studies tended to report
reductions in postnatal weight of offspring exposed to Pb; however, recent literature is inconsistent. Some
studies reported reductions of offspring weight following exposure to Pb in prenatal or early postnatal
life, while others report no effects of Pb on postnatal weight in offspring. Discerning reasons for the
observed inconsistencies is difficult because studies still reported results that contrasted with other studies
that used similar dosing windows, doses, and animal species.

8.4.2 Effects on Puberty among Females

The recent epidemiologic and toxicological studies examining the relationship between Pb
exposure and effects on puberty among females are summarized in the text below. Study details of the
recent epidemiologic studies are included in Table 8-12.

8.4.2.1 Epidemiologic Studies on Puberty among Females

The epidemiologic studies reviewed in the 2013 Pb ISA found consistent associations between
higher concurrent blood Pb and delayed pubertal development in females. The association persisted in
populations with mean and/or median concurrent BLLs of 1.2-9.5 (ig/dL. While most of the studies had
large sample sizes and controlled for potential confounders, they were cross-sectional study designs, so
there are some uncertainties regarding temporality between Pb exposure and pubertal onset; additionally,

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these studies were not able to separate out the influence of past Pb exposure, including prenatal
exposures, from more recent exposures.

The recent epidemiologic studies assessing the associations between blood Pb and onset of
puberty among females used different markers of puberty. A single cross-sectional study of NHANES
(2011-2012) data evaluated the associations between blood Pb concentration and circulating serum total
testosterone levels in 6-19-year-old children and adolescents (Yao et ah. 20.1.9'). Testosterone is a
principal sex hormone needed for normal physiologic processes during all lifestages and for females,
testosterone is of crucial importance for bone density and necessary for normal ovarian and sexual
function, libido, energy, and cardiovascular and cognitive functions ("Yao et ah. 20.1.9). While there were
no associations between blood Pb and testosterone in female children (6-11 years), serum testosterone
levels were 14.85% greater (95% CI: 0.83%, 30.81%) in female adolescents (12-19 years) in the lowest
quartile of BLLs (<0.35 (ig/dL) than those in the highest quartile (>0.63 (ig/dL) in the fully adjusted
model. For both female children and adolescents, there were no significant trends with increasing
quartiles of exposure (p for trend 0.63 and 0.08, respectively).

In a cross-sectional study in Poland, two different groups of adolescent girls aged 7-16 years
(n = 436 in 1995 and n = 361 in 2007) were assessed for effects of Pb on the age at attaining menarche
("Slawinska et ah. 2012). While the associations between blood Pb and menarche from either group of
girls were null (1995 OR: 0.70 [95% CI: 0.27, 1.85] and 2007 OR: 0.31 [95% CI: 0.09, 1.06]), the
patterns of association between these two times periods suggest delayed menarche. In another cross-
sectional study among school-age girls (n = 490) in Poland, there was a pattern of decreased odds
between blood Pb and age at menarche, whether controlled for BMI (OR: 0.54 [95% CI: 0.26, 1.13]),
percentage of body fat (OR: 0.52 [95% CI: 0.25, 1.08]), or sum of skinfolds (OR: 0.53 [95% CI: 0.26,
1.10]) (Goroula et ah. 2022). While these cross-sectional studies reported imprecise associations, the
pattern of association is important to note.

Three successive, cross-sectional Flemish Environment and Health Studies (FLEHS I, FLEHS II,
and FLEHS III) were conducted among adolescents (aged 14-15 years old) in Belgium between 2002-
2015 (De Craemer et ah. 2017). Female puberty markers of age at menarche, breast development, and
pubic hair development were evaluated in relation to blood Pb exposure. There was a consistent pattern of
delayed age at menarche across the three study cohorts (FLEHS I OR: 0.039 [95% CI: -0.072, 0.15];
FLEHS II OR: 0.257 [95% CI: 0.091, 0.424]; FLEHS III OR: 0.126 [95% CI: -0.021, 0.273]). The
associations between blood Pb and breast development were inconsistent, but there was indication of
delayed development among FLEHS I participants (OR: 0.798 [95% CI: 0.653, 0.969]). There were no
associations between blood Pb and development of pubic hair among adolescent females across the three
study cohorts.

Multiple cohort studies examined the associations between Pb exposure and puberty in females.
These studies used different biomarkers of exposure and different markers of puberty (Liu et ah. 20.1.9b:
Jansen et ah. 20.1.8: Nkomo et ah. 20.1.8). Cord BLLs and BLLs at age 13 were evaluated in association

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with puberty progression (development of pubic hair and development of breasts) among 684 females in
the Birth to Twenty Plus (BT20+) birth cohort in South Africa (Nkomo et aL 20.1.8"). In females with
elevated BLLs (>5 (ig/dL) at age 13, there was lower level of breast development (RR: 0.45 [95% CI:
0.29, 0.68]) and slower progression of pubic hair development (RR: 0.46 [95% CI: 0.27, 0.77]), but there
were no associations between cord blood Pb and pubic hair or breast development at age 13. In a cohort
of Mexican children, cumulative blood Pb from 1-4 years old was associated with delayed breast
development (OR: 0.96 [95% CI: 0.92, 0.99]) and delayed pubic hair development (OR: 0.95 [95% CI:
0.92, 0.99]), but maternal patella Pb and maternal tibia Pb were not associated with either breast or pubic
hair development in girls between 9-18 years old (n = 283) (Liu et aL. 2019b'). Additionally, the highest
tertile of maternal patella Pb and the second tertile of cumulative blood Pb from 1-4 years of age was
associated with delayed menarche. In a subset of the ELEMENT project cohort (n = 200), maternal blood
was measured during each trimester of pregnancy and daughters (mean age at follow-up assessment
13.8 ± 2.0 years) were asked about the occurrence of their first menstrual cycle ("Jansen et al.. 2018). Only
second trimester maternal BLLs were associated with later age at menarche (HR: 0.59 [95% CI: 0.28,
0.90]).

8.4.2.2	Toxicological Studies on Puberty among Females

There were no recent animal toxicological studies on the effects of Pb on puberty in females. The
2013 Pb ISA reported that one research group lavicoli et al. (lavicoli et al.. 2006; lavicoli et al.. 2004)
observed that mouse offspring exposed to Pb prior to birth and through puberty (BLLs 0.7-13 (ig/dL)
resulted in a dose-dependent delay in multiple markers of sexual maturation (e.g., vaginal opening, age at
first estrus). The latter study by this group utilized a multigenerational dosing paradigm in which they
observed delays of pubertal onset in the F2 generation similar to those seen in the Fi. The 2013 Pb ISA
also summarized a study in which Fisher 344 rats were dosed daily via gavage (12 mg/mL Pb) starting
30 days prior to breeding through weaning of the offspring (PND 23), resulting in delayed age at vaginal
opening in the offspring (BLLs of dams just prior to breeding averaged 39.8 (.ig/dL) (Pine et aL. 2006). Of
particular interest is that the observed delay in vaginal opening was attenuated in offspring that received
IGF-1 injections starting on PND 28 until vaginal opening was observed, demonstrating that IGF-1 is a
critical element to Pb-induced pubertal onset delays. Reports of delayed puberty due to Pb exposure in the
2013 Pb ISA are consistent with studies in the 2006 Pb AQCD which observed delays in puberty in
female Fisher 344 and Sprague-Dawley rats exposed to Pb during gestation and/or lactation (Dearth et al..
2004; Dearth et al.. 2002; Ron is et al.. 1.998c; Ronis et al.. .1.996).

8.4.2.3	Integrated Summary of Effects on Puberty among Females

There were several markers of puberty among females that were assessed for associations with Pb
exposures in the recent epidemiologic studies. Multiple cross-sectional studies and a single cohort study

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reported an association between blood Pb and delayed menarche among females, with relevant BLLs
("Gomula et ah. 2022; Jansen et ah. 20.1.8; De Craemer et ah. ; lawinska et ah. 20.1.2"). While these
cross-sectional studies reported imprecise associations, the pattern of association is important to note.
There was also indication of slower breast development, but the studies assessing breast development
were limited (Nkomo et ah. 20.1.8; De Craemer et ah. 2017). Additionally, there were a limited number of
studies that evaluated the development of pubic hair, but these results were inconsistent (Liu et ah. 20.1.9b;
Nkomo et ah. 20.1.8; De Craemer et ah. 2017). A single study among NHANES participants reported
increased serum total testosterone levels in female adolescents. The recent epidemiologic studies
assessing the associations between Pb exposure and puberty among females were limited by the timing of
the exposure to Pb and biomarker of exposure (blood, maternal bone, cord blood). However, these studies
consider a wide range of confounders, including height, weight, and BMI, and the associations reported
demonstrate that Pb exposure could result in physiological responses that contribute to changes in puberty
in females.

No recent PECOS-relevant toxicological studies investigated the effects of Pb on puberty.
However, studies from the 2013 Pb ISA and the 2006 Pb AQCD provide toxicological evidence that
indicates that Pb delays onset of puberty in female rodents. Several studies report delays in pubertal
markers such as vaginal opening and first estrus. Of note is that one study. Pine et ah (2006) reported that
the observed delay in vaginal opening in Pb-treated animals was attenuated when Pb treated animals were
supplemented with IGF-1 starting on PND 28. This strongly suggests that the mechanism through which
Pb induces delays pubertal onset in females is dependent on IGF-1 disruption.

8.4.3 Effects on Puberty among Males

The recent epidemiologic and toxicological studies examining the relationship between Pb
exposure and effects on puberty among males are summarized in the text below. Study details of the
recent epidemiologic studies are included in Table 8-12.

8.4.3.1 Epidemiologic Studies on Puberty among Males

The epidemiologic studies reviewed in the 2013 Pb ISA demonstrated an inverse relationship of
Pb on pubertal development among males at low blood Pb (mean and/or median BLLs of 3.0 to
9.5 (ig/dL). The studies were mostly cross-sectional, but the findings from these studies were supported
by those from a prospective longitudinal study (Williams et ah. 20.1.0). Boys with higher (>5 (.ig/dL) BLLs
at ages 8-9 years old had 24% to 31% reduction of pubertal onset based on testicular volume (TV),
genitalia staging, and pubic hair staging (Williams et ah. 20.1.0). While temporality of effects is difficult to
establish due to the nature of the cross-sectional study design, the larger studies controlled for potential

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confounders, with a few studies considering the inclusion of dietary factors, but did not control for other
metal exposures that may impact the associations.

In an NHANES (2011-2012) analysis, concurrent BLLs and serum total testosterone levels were
measured in 6-19-year-old children and adolescents (Yao et ah. 20.1.9'). Testosterone is a principal sex
hormone needed for the normal physiologic processes during all life stages. In males, testosterone is
essential for the development and maintenance of secondary sexual traits, and can also influence bone
mass, muscle strength, mood, and intellectual capacity. When comparing the highest quartile of blood Pb
to the lowest, there was no association between serum testosterone levels in either male children ((3:
-13.09% [95% CI: -34.45%, 15.22%]) or male adolescents ((3: 6.32% [95% CI: -14.62%, 32.4%]).
Among three successive, cross-sectional Flemish Environment and Health Studies (FLEHS I, FLEHS II
and FLEHS III) of adolescents in Belgium between 2002-2015, blood Pb was negatively associated with
free estradiol (fE2; OR: 0.908 [95% CI: 0.839, 0.983]) and free testosterone (OR: 0.909 [95% CI: 0.828,
0.997]) in FLEHS II, but not in FLEHS I (De Craemer et ah. 20.1.7). The associations between blood Pb
and estradiol (E2), testosterone, sex hormone binding globulin (SHBG), luteinizing hormone (LH), and
follicle stimulating hormone (FSH) were generally null (see Table 8-12). In addition to sex hormones, De
Craemer et ah (20.1.7) also evaluated the associations between blood Pb and genital development and
pubic hair development among male adolescents. Across the three cross-sectional studies, there was
decreased odds of delayed onset of genital development (FLEHS I OR: 0.843 [95% CI: 0.717, 0.99];
FLEHS II OR: 0.697 [95% CI: 0.462, 0.998]; FLEHS III OR: 0.621 [95% CI: 0.388, 0.967]) and pubic
hair development (FLEHS I OR: 0.808 [95% CI: 0.686, 0.949]; FLEHS II OR: 0.849 [95% CI: 0.563,
1.365]; FLEHS III OR: 0.515 [95% CI: 0.327, 0.774]).

In the BT20+ birth cohort in South Africa, cord BLLs and blood levels at age 13 were evaluated
in association to puberty progression pubic hair development and genital development in 732 males
(Nkomo et ah. 2018). In males, elevated cord BLLs (>5 (.ig/dL) was associated with slower pubic hair
development (RR: 0.28 [95% CI: 0.11, 0.74]). There were no associations between cord blood Pb and
genital development or BLLs at age 13 with pubic hair development or genital development. Similarly,
there were no associations between maternal patella Pb, maternal tibia Pb, and cumulative blood Pb from
1 -4 years old and pubertal development (genitalia, pubic hair, and TV) in boys ("Liu et ah. 2019b').

However, in a longitudinal cohort of boys from the Russian Children's Study, higher BLLs
(>5 (ig/dL) measured at age 8-9 years old (baseline) had pubertal onset 7.7-8.4 months later, on average,
than those with lower BLLs (<5 (.ig/dL) (Williams et ah. 20.1.9). Boys with higher BLLs at baseline had
later adjusted mean age at sexual maturity, with 4.2-5.1 months later attainment compared to boys with
lower BLLs. There was a shift in mean age for age at pubertal onset for stage 2 genitalia (G2) of
8.40 months (95% CI: 3.70, 13.10), 8.12 months (95% CI: 3.46, 12.78) for stage 2 pubic hair (P2), and
7.68 months (95% CI: 3.46, 11.90) for TV >3 mL. There was a shift in mean age for age at sexual
maturity for stage 5 genitalia (G5) of 4.20 months (95% CI: 0.56, 7.84) and 5.14 months (95% CI: 1.70,
8.58) for TV >20 mL, but a null association for stage 5 pubic hair (4.23 months [95% CI: -0.31, 8.77]).

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Furthermore, there was no shift in the mean age for duration of pubertal progression for genitalia (G2 to
G5), pubic hair (P2 to P5), or TV (>3 mL to >20 mL). In a mediation analysis, growth measurements at
age 11 were included to better understand what portion of the shift in mean age at sexual maturity was
attributable to the effect of BLL on growth. The association of peripubertal BLL with height Z-score
(HTZ) at age 11 accounted for 34%-53% of the total effect of BLLs on age at maturity, while BMIZ-
score at age 11 only accounted for 6%-23%. In another Russian Children's Study, Fleisch et al. (20.1.3)
longitudinally measured serum insulin-like growth factor (IGF-1) to assess the association with childhood
BLLs. BLLs were measured at baseline only and IGF-1 levels were only measured during the follow-up
periods. Boys were enrolled between the ages of 8-9 years and were prospectively followed, with IGF-1
measurements obtained at two-year follow-up (ages 10-11 years) and at four-year follow-up (ages 12-
13 years). The overall mean IGF-1 concentration was 29.2 ng/mL lower (95% CI: -43.8, -14.5) for boys
with high BLLs at age 8-9 years (>5 (ig/dL [max is 31 |ig/dL|) versus those with lower baseline BLLs
(<5 (ig/dL) in adolescence among boys.

8.4.3.2	Toxicological Studies on Puberty among Males

There were no recent toxicological studies on the effects of Pb on puberty in males, as was also
the case at the time of the 2013 Pb ISA. The 2006 Pb AQCD reported that one study found that prenatal
exposure to Pb delayed sexual maturation in a dose-dependent manner in male rats ("Ronis et al.. 1998c).
Specifically, Ronis et al. (1998c) reported that prostate weight (used in this study as a marker of sexual
maturation) was reduced in male rat offspring treated with Pb from GD 5 through sacrifice.

8.4.3.3	Integrated Summary of Effects on Puberty among Males

The epidemiologic studies reviewed in the 2013 Pb ISA demonstrated an inverse relationship of
Pb on pubertal development among males at low blood Pb (mean and/or median BLLs of 3.0 to
9.5 (ig/dL). Overall, the recent epidemiologic studies assessing the associations between Pb exposure and
different markers of puberty (hormone levels, pubic hair development, genital development, TV) among
males reported more inconsistent findings at low BLLs. Additionally, there were differences in the timing
of exposure to Pb and different biomarkers of Pb exposure (maternal blood, maternal bone, cord blood, or
concurrent blood). The recent epidemiologic studies were able to consider a wide range of confounders,
including height, weight, and BMI, and some studies were conducted among established longitudinal
cohorts. No recent toxicological studies reported on the effects of Pb on male puberty. Similarly, the 2013
Pb ISA reported no studies investigating Pb and male puberty. One study reported by the 2006 Pb AQCD
investigated the effects of Pb on puberty using prostate weight as a marker of sexual maturity in male rats
exposed to Pb starting on GD 5. Treatment with Pb resulted in reductions in prostate weight around the
time of puberty, possibly indicating that Pb delayed the onset of sexual maturation in Pb-treated animals
when compared to control.

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8.4.4

Other Developmental Effects

There were several recent studies that evaluated associations between Pb exposure and other
developmental effects in the epidemiologic and toxicological literature. Study details of the recent
epidemiologic studies are included in Table 8-13 and the recent toxicological studies are in Table 8-11.

8.4.4.1	Epidemiologic Studies on Other Developmental Effects

There were several recent studies with other outcomes related to developmental effects. In studies
of other related to developmental effects, there was a negative association with child blood Pb and
mitochondrial DNA copy number (Alegria-Torres et aL 2020); positive associations with maternal Pb
exposure and diurnal Cortisol rhythms in infants (Tamavo v Ortiz et aL 2016'); and lower salivary sialic
acid levels (a metric for oral anti-inflammatory potential which may increase the risk of dental caries)
("Hon et aL. 2020). but no associations between tooth Pb levels (second trimester, third trimester, or
postnatal) and alpha diversity metrics (bacterial or fungal), indicators of gut microbiota ("Sitarik et aL.
2020) or child blood Pb and telomere length ("Alegria-Torres et aL. 2020).

8.4.4.2	Toxicological Studies on Other Developmental Effects

The 2013 Pb ISA and the 2006 Pb AQCD did not report any studies that investigated the effects
of Pb exposure on developmental milestones. However, some recent studies have investigated these
outcomes (Table 8-11). One study dosed Wistar dams via drinking water (0.2% Pb) either prior to
conception, during gestation only, during lactation only, or during both gestation and lactation and
reported that only exposure during both gestation and lactation elicited impacts on developmental
milestones (Rao Barkur and Bairv. 20.1.6). Specifically, the age at eye opening was reduced. However,
although this exposure paradigm was the only one that produced effects on age at eye opening, it was also
the only paradigm that resulted in BLLs higher than 30 (ig/dL and reported a BLL of 31.59 (ig/dL in pups
on PND 22. Rao Barkur and Bairv (20.1.6) also investigated other developmental milestones such as pinna
detachment and tooth eruption but reported no Pb-induced changes in either of these outcomes. An
additional study investigated the effects of Pb on similar outcomes including eye opening, eye slit
formation, fur development, tooth eruption, and pinna detachment, but reported no effects when Wistar
dams were dosed via drinking water (0.2% Pb) from GD 6 to 21 (BLLs 11.2 (ig/dL in pups on PND 21)
(Basha and Reddv. 2015).

8.4.4.3	Integrated Summary of Other Developmental Effects

The recent epidemiologic studies have the potential to provide initial support of potential
mechanistic pathways for diurnal Cortisol rhythms, lower salivary sialic acid levels, and DNA oxidative

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stress damage from Pb exposure among children during developmental periods. However, the small
number of studies limits the ability to judge coherence and consistency across the outcomes evaluated in
these studies, although the associations with diurnal Cortisol rhythms, lower salivary sialic acid levels, and
decrease in mitochondrial DNA copy number indicate that Pb exposure could result in physiological
responses that may contribute to adverse developmental effects. Recent toxicological studies on other
developmental effects of Pb largely pertain to the effects of Pb on developmental milestones of offspring.
Of the few toxicological studies available, no effects of Pb on developmental milestones were reported
with the only exception being a reduction at the age of eye opening, but this treatment group had BLLs
higher than 30 (ig/dL.

8.5 Effects on Female Reproductive Function

The 2013 Pb ISA concluded that the relationship observed with female reproductive outcomes,
such as fertility and hormone levels in some epidemiologic and toxicological studies was sufficient
evidence to conclude a suggestive causal relationship between Pb exposure and female reproductive
function. Epidemiologic studies provided information on different exposure periods and support the
conclusion that Pb possibly affects at least some aspects of female reproductive function. However,
toxicological studies were less supportive for suggesting a causal relationship between Pb exposure and
female reproductive function. This may primarily be due to a lack of variety in female reproductive
endpoints investigated by studies identified in the literature. The only outcomes reported by PECOS-
relevant toxicological studies include litter size, number of litters, and maternal body weight.
Subsequently, no evidence was available for outcomes such as cyclicity, female hormones, sex organ
histopathology (including ovarian follicular counts), or female fertility indicators (e.g., latency to
pregnancy, implantation counts, conception rate). Additionally, the available toxicological evidence was
inconclusive, and the only studies that reported effects on female reproductive outcomes also reported Pb-
induced reductions in brain weight, indicating the possibility that animals were experiencing overt
toxicity from Pb ("Saleh et ah. 20.1.9; Sal eh et ah. 2018). The recent epidemiologic and toxicological
studies of Pb exposure and female reproductive function are detailed in the following sections.

8.5.1 Effects on Hormone Levels and Menstrual/Estrous Cycle

The recent epidemiologic and toxicological studies examining the relationship between Pb
exposure and hormone levels and menstrual/estrous cycle are summarized in the text below. Study details
of the recent epidemiologic studies are included in Table 8-14 and the recent toxicological studies are in
Table 8-15.

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8.5.1.1 Epidemiologic Studies on Hormone Levels and Menstrual/Estrous Cycle

The epidemiologic studies reviewed in the 2013 Pb ISA reported associations between
concurrent/closely timed BLLs and hormone levels in female adults. However, while there were changes
in hormone levels, there were inconsistencies in the hormones that were evaluated across the different
studies. A limitation of some the epidemiologic studies evaluated was the cross-sectional design, which
leaves uncertainty regarding Pb exposure magnitude, timing, duration, and frequency that contributed to
the observed associations. Additionally, the covariates included in statistical models as potential
confounders varied among studies, which could contribute to between study heterogeneity. Another
limitation of the epidemiologic studies is that not all of the studies investigated important confounders,
such as other metal exposures or smoking. The recent epidemiologic studies are divided into studies on
hormone levels and studies on menstrual/estrous cycle. The recent epidemiologic studies on hormone
levels in the following section are specific to hormones related to reproductive function and recent
epidemiologic studies on other hormones are described in Section 9.4.2 in the Other Health Effects
Appendix.

8.5.1.1.1	Epidemiologic Studies on Hormone Levels in Females

There were a few cross-sectional studies that evaluated the associations between Pb exposure and
different hormones in females (Lee et ah. 20.1.9; Chen et ah. 20.1.6; Kriee and Feng. 20.1. IV These studies
used population-based surveys to evaluate associations between blood Pb and hormones and found
consistent positive associations with FSH in post-menopausal women. In an NHANES (1999-2002)
analysis, Krieg and Feng (20.1.1.) evaluated serum FSH and LH. Serum FSH slope increased per every
logio-blood Pb increase in the post-menopausal women ((3: 26.38 [95% CI: 13.39, 39.38]), women who
had both ovaries removed ((3: 27.71 [95% CI: 1.64, 53.78]), and pre-menopausal women ([3: 11.97 [95%
CI: 3.27, 20.66]), but serum FSH was not associated with BLLs in pregnant women, women who were
menstruating, or women who were taking birth control pills. Serum LH slope increased per every logio-
blood Pb increase in the post-menopausal women ([3: 11.63 [95% CI: 4.40, 18.86]) and women who had
both ovaries removed ([3: 20.59 [95% CI: 2.14, 39.04]), but serum LH was not associated with BLLs in
the pregnant women, women who were menstruating, women who were taking birth control pills, and
pre-menopausal women. In another cross-sectional, population-based survey in China. Chen et ah C
examined the associations between blood Pb and total testosterone (tT), E2, and SHBG, in addition to LH
and FSH in postmenopausal women (age >55 years). When comparing the highest quartile of blood Pb
(>5.98 (ig/dL) to the lowest (<2.70 (.ig/dL). there were positive associations with BLLs and SHBG ([3:
0.048, SE: 0.016, p < 0.01), FSH ([3: 0.046, SE: 0.016, p < 0.01), and LH ([3: 0.037, SE: 0.016, p < 0.05).
There were null associations between BLLs and tT or E2. Across the quartiles of blood Pb, there were
also positive trends observed with SHBG (p for trend: 0.002), FSH (p for trend: 0.001), and LH (p for
trend: 0.026), suggesting a potential linear exposure response between blood Pb and these hormones. In a
study of the Korea National Health and Nutrition Examination Survey (KNHANES) (2012-2014), blood

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Pb and serum FSH levels were assessed in postmenopausal women (aged 50 or older) (Lee et ah. 20.1.9').
Serum FSH levels were positively associated with increasing blood log-Pb ((3: 2.929 [95% CI: 0.480,
5.377]).

8.5.1.1.2	Epidemiologic Studies on Menstrual/Estrous Cycle

There were no available epidemiologic studies in the 2013 Pb ISA that evaluated Pb exposure
with menopause. There are a limited number of studies that examined the relationship between Pb
exposure and menopause, and these recent studies reported consistent positive associations. A recent
cross-sectional study, NHANES (1999-2010) data was used to examine the associations between blood
Pb and menopause among women aged 45-55 years (age range where menopause is likely to occur)
(Mendola et aL 20.1.3). In the overall study sample (NHANES 1999-2010), with increasing quartiles of
blood Pb, there were increasing odds of menopause. Comparing the lowest BLLs (<1.0 |ig/dL). the odds
for Q2 through Q4 were 1.7 (95% CI: 1.0, 2.8), 2.1 (95% CI: 1.2, 3.6), and 4.3 (2.6, 7.2), respectively.
When adjusting for bone measurements (either bone alkaline phosphatase or femoral neck bone density),
the associations were similar. In a subset (n = 434) of the Nurse's Health Study, the associations between
bone Pb levels and age at menopause were explored (Euro et aL 20.1.4). Compared with women in the
lowest tertile of tibia Pb (<6.5 |ig/g). those in the highest tertile (>13 jxg/g) were 1.21 years younger at
menopause on average (95% CI: -2.08, -0.35; p for trend: 0.006). Women in the highest tertile of tibia
Pb had an increased odds of 5.30 (95% CI: 1.42, 19.78; p for trend: 0.006) for early menopause
(menopause before age 45) compared with women in the lowest tertile. The associations with early
menopause were null across tertiles for patella Pb and BLLs.

8.5.1.2 Toxicological Studies on Hormone Levels and Menstrual/Estrous Cycle

There are no recent animal toxicological studies on the effects of Pb on the menstrual/estrous
cycle. The 2013 Pb ISA did not report any studies that investigated the effects of Pb on the
menstrual/estrous cycle, however some studies were summarized in the 2006 Pb AQCD. Specifically,
studies conducted in non-human primates found that Pb exposure increased menstrual cycle variability,
reduced days of menstrual flow, increased cycle length, and reduced progesterone ("Franks et aL. .1.989;
Laughlin etah. .1.987). However, another study with a lower BLL than the previous studies (<40 (.ig/dL
versus 44-89 (ig/dL) did not report an effect on the menstrual cycle in a non-human primate species
(Foster etah. .1.992). Impacts of Pb on estrous cyclicity were examined in two rat studies that both utilized
multiple dosing paradigms to assess the varying impacts Pb exposure may have during different
developmental periods. Specifically, one study used the following exposure windows: gestation only,
lactation only, gestation and lactation, postnatal (from birth and continued past weaning), and continuous
(from the beginning of gestation continued past weaning) (Ron is et aL. 1998a). This study reported that
offspring in the postnatal and continuous exposure groups had fewer females who were regularly cycling.

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The other study was conducted by the same research group and utilized the following dosing windows:
post-pubertal (PND 60 to PND 74), pre-pubertal (PND 24 to PND 74), and in utero (GD 5 to PND 85)
(Ronis et ah. 1996V Rats in the pre-pubertal and in utero exposure paradigm groups experienced estrous
cyclicity disruption. While this study seems to indicate that pre-pubertal periods are more sensitive to
chemical insult, the previous study by Ron is et al. (1998a) suggests that normal cyclicity is recoverable
after cessation of exposure. However, it is worth noting that in both of these studies the BLLs were very
high, with a range of 63.2-264 (ig/dL for treatment groups that displayed treatment-related effects.

There are no recent animal toxicological studies on the effects of Pb on reproductive hormones.
The 2013 Pb ISA reported on a few studies that investigated the effects of Pb on reproductive hormones,
but none on cyclicity. Rodent studies reported that gestational and lactational exposure decreased
circulating levels of progesterone and E2 (Pillai et al.. 20.1.0; Nampoothiri and Gupta. 2008). Dumitrescu
et al. (2008b) reported similar findings in adult female Wistar rats that were exposed to Pb for 6 months
via drinking water. Dumitrescu et al. (2008b) reported reductions in E2, progesterone, and FSH and
increases in LH and testosterone. The 2006 Pb AQCD reports findings from some toxicological studies
that show effects of Pb on hormones and cyclicity. Reductions in progesterone were observed in a study
wherein monkeys had BLLs of 25 to 30 (ig/dL, but no such reductions in progesterone were observed in
monkeys with even lower BLLs (10 to 15 (.ig/dL) (Foster et al.. .1.996).

8.5.1.3 Integrated Summary of Effects on Hormone Levels and Menstrual/Estrous
Cycle

The recent cross-sectional, population-based survey epidemiologic studies found consistent
positive associations between blood Pb and FSH in women who were post-menopausal. While these
studies are limited by their study design, the studies were conducted in well-established population-based
surveys. These studies considered a range of confounders, including controlling for BMI and smoking,
even co-exposure to other metals. The recent studies examining the relationship between menopause and
Pb exposure found consistent positive associations. The results from concurrent exposure of blood Pb
with menopause were supported by the results from a longitudinal cohort that examines bone Pb, a
cumulative biomarker of Pb exposure, and menopause, both the difference in age at menopause and risk
of early menopause. No recent PECOS-relevant toxicological studies reported on the effects of Pb on
hormone levels in females or menstrual or estrous cyclicity. However, previous toxicological evidence
suggests that Pb may disrupt reproductive hormones and menstrual and estrous cyclicity in females. Two
toxicological studies in rats reported disruptions in estrous cyclicity, and two toxicological studies based
in non-human primates reported alterations to different menstrual cycle aspects (e.g., length of cycle,
length of menstruation) and reproductive hormone levels. Additional rodent studies reported effects of Pb
on circulating reproductive hormone levels, including sex steroid hormones (progesterone, testosterone,
and E2) and gonadotropin hormones (LH and FSH).

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8.5.2

Effects on Female Fertility

Multiple epidemiologic and toxicological studies have examined the relationship between Pb and
female fertility. These studies are summarized in the text below. Study details of the recent epidemiologic
studies are included in Table 8-14 and the recent toxicological studies are in Table 8-15.

8.5.2.1 Epidemiologic Studies on Female Fertility

The epidemiologic studies reviewed in the 2013 Pb ISA examined a variety of fertility-related
endpoints. Although some studies demonstrated an association between higher Pb biomarker levels and
fertility/pregnancy, the results are inconsistent across studies. One limitation in most of these studies is
that the participants were women seeking help for fertility problems. The participants were not samples of
the general population and therefore cannot be generalized to all women of childbearing age. This may
also have introduced substantial selection bias into the study.

The recent epidemiologic studies also evaluated different outcomes to measure fertility. In an
NHANES (2013-2014 and 2015-2016) study, Lee et al. (2020) assessed whether BLLs were associated
with self-reported infertility by comparing BLLs of infertile women (n = 42) to pregnant women (n = 82).
There was increased risk of 2.60 (95% CI: 1.05, 6.41) per two-fold increase in BLLs of infertility. When
BLLs were categorized into tertiles, risk of infertility was more pronounced (OR: 5.40 [95% CI: 1.47,
19.78] interfile 2 (0.41-0.62 ^ig/dL) and OR: 5.62 [95% CI: 1.13, 27.90] interfile 3 (0.63-5.37 ^ig/dL),
respectively). In the LIFE Study, a cohort of couples were followed prospectively to assess persistent
environmental chemicals and human fecundity (Louis et al.. 20.1.2). BLLs in both the female and male
partners were collected at baseline. Female BLLs were not associated with increased time to pregnancy in
the female exposure model (OR: 0.97 [95% CI: 0.85, 1.11]) or the couple exposure model (OR: 1.06
[95% CI: 0.91, 1.24]). However, there was decreased odds, or longer time to pregnancy, for male BLLs in
both the male exposure model (OR: 0.85 [95% CI: 0.73, 0.98]) and the couple exposure model (OR: 0.82
[95% CI: 0.68, 0.97]).

In a cross-sectional study among infertile women in Taiwan, Lai et al. C examined the
associations between BLLs and diagnosis of endometriosis, which can cause infertility. Increasing tertiles
of BLLs was associated with higher odds of endometriosis (OR: 2.59 [95% CI: 1.11, 6.06] for T3
compared to Tl). In a cohort study, among couples undergoing the first in vitro fertilization (IVF) cycle,
maternal Pb levels were assessed with pregnancy outcomes (Li et al.. 2022). Pb levels in serum were
collected before oocyte retrieval. With increasing maternal serum Pb levels, there was a reduction in
successful implantation (OR: 0.85 [95% CI: 0.77, 0.94]) and clinical pregnancy (OR: 0.95 [95% CI: 0.91,
0.99]). Additionally, when maternal serum Pb was categorized into tertiles, there was a lower rate of
successful implantation (OR: 0.58 [95% CI: 0.40, 0.85]) in the highest Pb tertile, compared to the lowest
Pb tertile. Among tertiles of Pb serum levels, the associations were null for clinical pregnancy.
Furthermore, there was a negative association with maternal serum Pb and high-quality embryo rate ([3:

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-0.14 [95% CI: -0.32, -0.04]), but there were null associations with all other embryo quality indicators
(Table 8-14). In a cohort of 195 couples undergoing IVF, Pb was measured in serum and follicular fluid
from the female partner and semen from the male partner in association with six IVF outcomes ("Zhou et
ah. 2021a'). There were no associations between serum or follicular fluid Pb levels and any IVF
outcomes—normal fertilization, good embryo, blastocyst formation, high-quality blastocyst, pregnancy,
or live birth.

8.5.2.2	Toxicological Studies on Female Fertility

The 2013 Pb ISA reported some studies that investigated the effects of Pb on female fertility. A
handful of these studies reported that exposure to Pb reduced litter sizes in exposed female rats
("Dumitrescu et ah. 2008a; Teiion et ah. 2006) and mice (Iavicoli et ah. 2006). Contrasting this is a study
that found no changes in fertility rate or litter size in female rats treated prior to mating through pregnancy
(Nampoothiri and Gupta. 2008). Recent studies corroborate the findings of Nampoothiri and Gupta
(2008) and do not demonstrate any effects of Pb on female fertility in terms of litter size or number of
litters produced by dosed dams in mice (Schneider et ah. 20.1.6; Corv-Slechta et ah. 20.1.3) or rats (Rao
Barkur and Bairv. 20.1.6; Barkur and Bairv. 20.1.5; Weston et ah. 20.1.4; Betharia and Mafaer. 20.1.2). Among
these recent studies, a variety of dosing paradigms were utilized, including exposure during
preconception, lactation, gestation, and combinations thereof (BLLs ranged between 3.02-26.86 (ig/dL in
pups on PND 2-22). The contrast in effects on litter size between studies that do and do not report effects
of Pb on litter size is perplexing, and the inconsistencies of BLL measurements (e.g., some measured Pb
levels in offspring, some measured in dams, some studies did not report BLLs at all) between studies
further exacerbates the difficulty of reconciling these contrasts. However, some plausible explanations for
these differences exist and primarily involve differences in study design. Studies that reported reductions
in litter size due to Pb exposure tended to either use higher doses (Teiion et ah. 2006). longer dosing
durations (Iavicoli et ah. 2006). or dosed sires in addition to dosing dams (Dumitrescu et ah. 2008a) when
compared to studies that did not report reductions in litter sizes.

8.5.2.3	Integrated Summary of Effects on Female Fertility

Among the recent epidemiologic studies, there were inconsistent associations between Pb
exposure and female fertility. In studies among participants in the general population, there was an
increased risk of self-reported infertility and longer time to pregnancy. However, among studies with
women who were either seeking help at a fertility clinic or reported infertility, the associations were
inconsistent. Because the study participants included only a small sample of women who were either
seeking help at a fertility clinic or reported infertility, there may be selection bias and limits the
generalizability of the results as study participants have already been diagnosed and are seeking treatment
for fertility issues. Further, pregnancy outcomes, such as successful implantation, are more likely to be

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ascertained from women seeking treatment at fertility clinics. Additionally, the recent epidemiologic
studies were limited by the concurrently measured exposure and outcome, different biomarkers of
exposure (e.g., blood, serum, and follicular fluid), and a small number of participants. However, these
studies did include adjustment for potential confounders, including age, BMI, and partner exposure.
Previous toxicological evidence reported inconsistent effects of Pb on fertility in females. All recent
toxicological studies reported that female fertility was not affected by Pb exposure, even when a variety
of dosing paradigms were used.

8.5.3 Effects on Morphology and Histology of Female Sex Organs (Ovaries,
Uterus, Fallopian Tubes/Oviducts, Cervix, Vagina, and Mammary
Glands)

Recent epidemiologic and toxicological studies evaluating the association between Pb exposure
and morphology or histology of female sex organs (ovaries, uterus, fallopian tubes/oviducts, cervix,
vagina, and/or mammary glands) are limited. Study details for the single cross-sectional epidemiologic
study are included in Table 8-14 and the toxicological studies are included in Table 8-15.

8.5.3.1	Epidemiologic Studies of Morphology and Histology of Female Sex Organs
(Ovaries, Uterus, Fallopian Tubes/Oviducts, Cervix, Vagina, and Mammary
Glands)

In the 2013 Pb ISA, there were no epidemiologic studies available that evaluated Pb
concentrations and associations with morphology or histology of female sex organs (ovaries, uterus,
fallopian tubes/oviducts, cervix, vagina, and/or mammary glands). A recent cross-sectional study
examined the association between BLLs and rate of uterine fibroids and uterine fibroid volume (Ye et aL
20.1.7'). Among 288 (46 with fibroids and 242 without) pre-menopausal women included in the study,
there were null associations between blood Pb and the presence of uterine fibroids (OR: 1.39 [95% CI:
0.75. 2.56]) and volume of the largest fibroids ((3: 0.12 [95% CI: -2.26, 2.51]). When blood Pb was
categorized into quartiles, the association with volume of uterine fibroids remained null. While the
associations between blood Pb and rate of uterine fibroids and uterine fibroid volume were generally null,
the women with uterine fibroids had higher geometric mean BLLs than women without fibroids
(1.43 (ig/dL versus 1.35 (ig/dL, respectively).

8.5.3.2	Toxicological Studies of Morphology and Histology of Female Sex Organs
(Ovaries, Uterus, Fallopian Tubes/Oviducts, Cervix, Vagina, and Mammary
Glands)

There are no recent animal toxicological studies on the effects of Pb on morphology or histology
of female sex organs. The 2013 Pb ISA discussed a single study that reported Pb exposure increased

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membrane fluidity in granulosa cells in Charles Foster rats that were dosed via intraperitoneal injections
for 15 days (Nampoothiri and Gupta. 2006). The 2006 Pb AQCD also reported that exposure to Pb during
early pregnancy caused structural changes in the uterine epithelium in mice (Nilsson etah. .1.991; Wide
and Nilsson. .1.979).

8.5.3.3 Integrated Summary of Morphology and Histology of Female Sex Organs
(Ovaries, Uterus, Fallopian Tubes/Oviducts, Cervix, Vagina, and Mammary
Glands)

There was a single recent epidemiologic study evaluating associations between Pb exposure and
uterine fibroids. Although this was a small cross-sectional study, it was able to control for a large range of
confounders. The results from this single study are limited by the small sample size and concurrent
measurements of blood Pb and fibroids. Toxicological evidence regarding Pb exposure and female sex
organs is scarce. No recent PECOS-relevant toxicological studies were available. Previous studies
discussed in the 2013 Pb ISA and the 2006 Pb AQCD were scarce and reported few effects of Pb on
female sex organ morphology or histology.

8.6 Effects on Male Reproductive Function

The 2013 Pb ISA concluded that there was toxicological evidence with supporting epidemiologic
evidence to conclude that a causal relationship exists between Pb exposure and effects on male
reproductive function. The key evidence was provided by toxicological studies in rodents, non-human
primates, and rabbits showing detrimental effects on semen quality, sperm, and fecundity/fertility with
supporting evidence in epidemiologic studies of associations between Pb exposure and detrimental effects
on sperm. Recently published research has continued to support an association between Pb and
sperm/semen production, quality, and function. Studies of Pb and male reproductive function are
described in the sections below.

8.6.1 Effects on Sperm/Semen Production, Quality, and Function

Multiple epidemiologic and toxicological studies have examined the relationship between Pb and
sperm and semen production, quality, and function. These studies are summarized in the text below.

Study details of the recent epidemiologic studies are included in Table 8-16 and the recent toxicological
studies are in Table 8-17. The majority of the recent epidemiologic studies are cross-sectional with
concurrent measurements of Pb levels in biological samples and sperm-related outcomes. Recent
toxicological studies use a variety of dosing paradigms, and those that dose for longer periods of time

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(>30 days) or during a developmental window most often reported effects of Pb exposure on aspects of
sperm and semen quality.

8.6.1.1 Epidemiologic Studies on Sperm/Semen Production, Quality, and Function

The 2013 Pb ISA epidemiologic studies of Pb exposure and sperm and semen production, quality,
and function were cross-sectional, mostly in occupational cohorts, with concurrent measurements of Pb
levels in biological samples and sperm-related outcomes. The multiple epidemiologic studies in
occupational cohorts had mean BLLs over 40 (ig/dL for individuals occupationally exposed to Pb. The
occupational studies also had limited consideration for potential confounding factors, such as other
workplace exposures, which may impact the associations. The epidemiologic studies of men attending
fertility clinics may be subject to selection bias, and the results may not be generalizable. Additionally,
these studies reported imprecise estimates, did not control for other potential confounders such as other
metals, and had small sample sizes.

Several recent cross-sectional studies have explored the relationship between Pb exposure and
sperm and semen production, quality, and function. These studies were all conducted in males attending
fertility clinics and reported inconsistent associations for various metrics of sperm/semen production,
quality, and function. There were other cross-sectional studies that also examined associations with sperm
and semen production, quality, and function using different and Pb measured in semen, seminal fluid, and
seminal plasma, but these findings were more inconsistent.

Among the cross-sectional studies that evaluated associations with blood Pb, there was lower
normal sperm morphology with increasing BLLs ("Shi et ah. 2021; Sukhn et al.. 20.1.8; Li et aL 20.1.5").
Additionally. Li et al. (20.1.5) reported increased odds of lower semen quality, sperm concentration,
numbers of sperm, total motility sperm, and progressive motility sperm with increasing BLLs, whereas
Sukhn et al. (20.1.8) reported null associations with sperm volume, concentration, total count, progressive
motility, viability, and World Health Organization (WHO) morphology, and Shi et al. (1 reported
null associations between blood Pb and semen parameters of semen volume, sperm concentration, total
sperm count, sperm motility, total motile sperm count, sperm vitality, DNA fragmentation index, and
percentage of acrosome reacted sperm (see Table 8-16). Furthermore, there were differences in BLLs
between men categorized as having low-quality semen and those classified as having normal or high-
quality semen (Sukhn et al.. 20.1.8; Li et al.. 20.1.5). Li et al. (20.1.5) reported mean blood Pb for men in the
low-quality semen group was 3.43 (ig/dL and 2.38 (ig/dL for those in the high-quality semen group, and
Sukhn et al. (20.1.8) reported the mean blood Pb for low-quality semen group of 5.198 (.ig/dL and
3.575 (ig/dL for the normal-quality semen group.

Other cross-sectional studies examined the relationship between Pb measured in seminal fluid and
metrics of sperm/semen production, quality, and function, but the associations were inconsistent (Jia et
al.. 2022; Sukhn et al.. 20.1.8; Pant et al.. 20.1.4). Pant et al. (20.1.4) measured Pb in semen and metrics of

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sperm/semen production, quality, and function, and reported associations of higher Pb levels in sperm
with decreased sperm motility ((3: -2.43% [95% CI: -4.87%, -0.001%]), decreased sperm concentration
((3: 1.97 106/mL [95% CI: -3.16, -0.33]), increased tail length ([3: 3.79 [95% CI: 0.56, 7.02]), increased
percent DNA in tail ([3: 1.31 [95% CI: 0.17, 3.74]), and increased tail movement ([3: 1.20 [95% CI: 0.23,
2.16]). However, Sukhn (	).1.8) assessed sperm characteristic relationships with seminal fluid Pb and

reported increased odds of below-reference sperm viability and WHO morphology with higher seminal
fluid Pb. but null associations with volume, concentration, total count, and progressive motility. Jiaet al.
(2022) reported no associations between seminal plasma Pb concentrations and semen parameters (semen
volume, sperm concentration, total sperm number, progressive motility, and normal morphological rate).

A recent cohort study examined the associations between peripubertal blood Pb, collected at
enrollment, and parameters of sperm and semen production, quality, and function for 223 participants in
the Russian Children's Study, with the semen sample collected 10 years after enrollment (Williams et al..
2022). There were null associations between peripubertal blood Pb and sperm parameters (sperm
concentration, total count, progressive motility, and total progressive motile sperm count, or probability
of having low semen quality based on sperm count/motility), whether blood Pb was modeled
continuously, categorized as tertiles, or categorized as low (<5 (ig/dL) blood Pb versus high (>5 (ig/dL)
blood Pb (see Table 8-16).

8.6.1.2 Toxicological Studies on Sperm/Semen Production, Quality, and Function

The 2013 Pb ISA summarized several toxicological studies that investigated the effects of Pb
exposure on sperm-related outcomes. Utilizing a variety of dosing paradigms and animal models,
previously published studies have demonstrated that Pb exposure reduced sperm counts, reduced numbers
of viable sperm, reduced motility, and increased morphological abnormalities (Filial et al.. 20.1.2; Anium
et al.. 20.1.1; Al louche et al.. 2009; Oliveira et al.. 2009; Salawu et al.. 2009; Shan et al.. 2009; Tapisso et
al.. 2009; Massanvi et al.. 2007; Piao et al.. 2007; Wang et al.. 2006). Results from recently published
studies tend to suggest that Pb exposure impacts sperm and semen parameters (Table 8-17). All available
studies that reported outcomes on sperm and semen parameters were conducted in mice. Only one study
utilized a developmental exposure paradigm and dosed lactating CD-I mice from PND 0 to 21 which
resulted in reduced numbers of sperm at PND 70 in male offspring in the highest dose group (BLLs
19.1 (.ig/dL) (Wang et al.. 2013a). Other studies that directly exposed male mice after weaning also
reported Pb-induced sperm alterations including increased incidence of abnormal morphology, reduced
density, and reduced viability (BLLs 9.4-11.92 (.ig/dL) (Zhang et al.. 2021; Xie et al.. 2020; Godinez-
Solis et al.. 20.1.9'). However, some studies also reported no Pb-induced effects on sperm motility,
concentration, count, or viability (BLLs 9.4-11.8 (ig/dL) (Pavlova et al.. 2021; Godinez-Solis et al..
20.1.9'). It is worth noting that while Pavlova et al. (202.1.) reported no Pb-induced effects on sperm count,
Pb-treated animals had sperm counts 25% lower than those of control mice, but this effect failed to reach
statistical significance (p = 0.146). In terms of patterns in the reported data, studies that utilized long-term

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exposure (>30 days) or dosed during developmental periods tended to report effects of Pb on sperm or
semen parameters, and Pavlova et al. (202.1.) was the only study to use short-term exposure during
adulthood and also was the only study to report no effects at all on any sperm or semen parameters.

8.6.1.3 Integrated Summary of Effects on Sperm/Semen Production, Quality, and
Function

Among the recent epidemiologic studies that evaluated associations between Pb exposure
(measured in blood, semen, seminal fluid, or seminal plasma) and effects on sperm/semen production,
quality, and function, there were inconsistent findings, which was similar to the conclusion in the 2013 Pb
ISA. More consistent associations were observed for blood Pb with decreased sperm/semen production,
quality, and function than for Pb measured in semen, seminal fluid, or plasma; however, there are
limitations in the recent epidemiologic studies. All the cross-sectional studies were conducted in males
attending fertility clinics, which may have resulted in selection bias and limits the generalizability of the
results. Further, the small sample size from these cross-sectional studies also reduces the statistical power
to determine the precision of the associations. With concurrent measurement of Pb exposure with
outcomes related to sperm/semen production, quality, and function, temporality cannot be established.
Lastly, the use of different biomarkers (e.g., blood, semen, seminal fluid, or seminal plasma) to measure
Pb exposure and the different metrics of sperm/semen production, quality, and function limits the ability
to judge coherence and consistency across studies. Despite these limitations, it is important to note that a
wide variety of potential confounders were controlled for, including hormone levels, which could
potentially impact sperm/semen production, quality, and function. Previous and recent toxicological
studies generally reported that Pb alters some aspect of sperm or semen quality, such as sperm density,
motility, morphology, and viability, especially those studies that employed dosing during developmental
periods or for periods 30 days or longer. All recent toxicological evidence was produced from mouse
strains, but previous toxicological studies report similar effects in other species such as rats and rabbits.

8.6.2 Effects on Hormone Levels in Males

The epidemiologic and toxicological studies reviewed in the 2013 Pb ISA reported inconsistent
results regarding changes in hormone levels in men and associations with Pb exposure. The results from
the 2013 Pb ISA were similar to the findings from the 2006 Pb AQCD. Recent epidemiologic and
toxicological studies are reported below. Epidemiologic studies were mostly cross-sectional with blood
Pb measured concurrently with hormone levels. Study details for the epidemiologic studies, including
BLLs, study population characteristics, potential confounder, and select results, are in Table 8-16.
Previous toxicological evidence regarding the effect of Pb on hormones in males is somewhat
inconsistent, but most studies reported impacts of Pb on hormone levels. Recent toxicological studies are
extremely limited, but support previous toxicological studies reported in the 2013 Pb ISA that observed

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Pb-induced effects on hormones in males. Study details for the recent toxicological studies are in
Table 8-17.

8.6.2.1 Epidemiologic Studies on Hormone Levels in Males

In the 2013 Pb ISA, there were a few epidemiologic studies that evaluated hormone levels in
males in association with Pb exposure. The findings of these studies were inconsistent. The epidemiologic
studies were limited by their sample populations, often occupational cohorts or men recruited from
fertility clinics, which may not be representative of the general populations and limits the generalizability
of the results. More specifically, the occupational cohorts may have other metal exposures that were not
considered and may confound the associations, while studies conducted with subjects from fertility clinics
are subject to selection bias. While these studies included important confounders such as smoking, other
factors, such as exposure to other metals, were often absent. The cross-sectional study design of some of
the epidemiologic studies reviewed makes it difficult for temporality of effects to be established.
Additionally, most of the epidemiologic studies examined concurrent Pb exposure and hormone levels,
which may not reflect changes resulting from long-term exposures.

The recent epidemiologic studies on hormone levels detailed in this section are specific to
hormones related to reproductive function and recent epidemiologic studies on other hormones are
described in Section 9.4.2 in the Other Health Effects Appendix.

There are a few recent epidemiologic cross-sectional studies that evaluated the associations
between hormone levels in males and Pb exposure. There were consistent positive associations between
blood Pb and testosterone among these studies. One NHANES analysis combined three consecutive
cycles of NHANES (1999-2000, 2001-2002, and 2003-2004) to investigate the associations between
blood Pb and various sex hormones: testosterone, free testosterone, E2, fE2, androstenedione glucuronide,
and SHBG among men over 20 years old (Kresovich et aL 2015). Comparing the highest quartile of
blood Pb exposure (>3.20 (ig/dL) to the lowest (<1.40 |ig/dL). testosterone was positively associated with
blood Pb ((3: 0.79, SE: 0.22) and there was an indication of exposure-response (p for trend: 0.0026). There
were null associations between blood Pb and all other sex hormones. In another NHANES (2011-2012)
study, blood Pb and serum testosterone were measured in men of reproductive age (18-55 years old)
(Lewis and Meeker. 2015). Of the 484 men included in the analysis, there was a 6.65% (95% CI: 2.09%,
11.41%) change in the serum testosterone concentration associated with a doubling (100% increase) in
blood Pb concentration. Chen et al. t also reported positive associations between concurrent
testosterone and blood Pb concentrations. Utilizing data from a population-based survey, the Survey on
the Prevalence in East China for Metabolic Diseases and Risk Factors (SPECT)-China, 2,286 men were
included in the analysis to investigate the relationship between quartiles of BLLs and multiple
reproductive hormones - tT, SHBG, E2, LH, and FSH. When comparing the highest quartile of blood Pb
(>6.249 (ig/dL) to the lowest (<2.90 |ig/dL). there were positive associations with BLLs and tT ((3: 0.033,

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SE: 0.010, p < 0.01), SHBG ((3: 0.038, SE: 0.012, p < 0.01), FSH ((3: 0.030, SE: 0.015, p < 0.05), and LH
((3: 0.028, SE: 0.013, p < 0.05), but null associations with E2 ((3: -0.003, SE: 0.017). Across the quartiles
of blood Pb, there were also positive trends observed with tT (p for trend: 0.012), SHBG (p for trend
<0.001), FSH (p for trend <0.001), and LH (p for trend <0.001), suggesting a potential linear
concentration response.

While there were consistent positive associations between blood Pb and serum testosterone in the
cross-sectional studies, a single cohort study reported null associations. Among a subset of participants
(n = 453) in the Russian Children's Study, there were no associations between peripubertal BLLs and
hormones levels (testosterone, LH, or FSH) measured between 8-19 years of age, whether blood Pb was
modeled continuously or categorized (<5 (.ig/dL versus >5 (.ig/dL) ("Williams et ah. 2022).

8.6.2.2	Toxicological Studies on Hormone Levels in Males

The 2013 Pb ISA discussed several studies that reported on the effects of Pb on hormone levels in
males. All studies were conducted in rats, and all directly dosed the tested animals save for one that
exposed gestating and lactating dams and measured hormones in offspring. Dosing durations varied from
21 days to 24 weeks, and most studies reported reductions in testosterone (Pillai et al.. 20.1.2; Aniuro et aL
2011: Biswas and Ghosh. 2006: Rubio et al.. 2006'). One study observed increased testosterone ("Allouche
et aL. 2009) and another reported reductions in LH and FSH (Biswas and Ghosh. 2006). However, not all
studies observed effects on male hormones due to Pb exposure. One study observed no change in
testosterone ("Salawu et al.. 2009) and another reported no change in FSH and LH levels despite reporting
increased testosterone levels ("Allouche et al.. 2009). Only one recent PECOS-relevant toxicological study
was published that investigated the effects of Pb exposure on hormones in males (Table 8-17). This study
dosed nursing CD-I mice from PND 0 to 21 and reported reduced serum testosterone at weaning and
PND 70 and reduced testicular testosterone at weaning in offspring in the highest dose group (19.1 (ig/dL)
(Wang et al.. 2013a).

8.6.2.3	Integrated Summary of Effects on Hormone Levels in Males

The recent cross-sectional epidemiologic studies reported consistent associations between blood
Pb and testosterone; however, a single cohort study reported no associations. Of note, the recent cross-
sectional studies were in adult men, whereas the single cohort study was in male adolescents.
Additionally, the study by Chen <	) .1.6) provides further support of a positive association with

SHBG, FSH, and LH and blood Pb. The positive trends among quartiles of blood Pb and testosterone,
SHBG, FSH, and LH provide insight on the possible concentration-response relationship. These studies
have robust sample sizes drawn from population-based surveys and controlled for a number of
confounders, including smoking, but the temporality of effects is difficult to establish due to the nature of

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cross-sectional study design. Recent toxicological evidence regarding the effects of Pb on hormones in
males is extremely limited, but in agreement with most studies summarized in the 2013 Pb ISA which
report effects of Pb on hormones in males.

8.6.3 Effects on Male Fertility

The recent epidemiologic and toxicological studies examining the relationship between Pb
exposure and male fertility are summarized in the text below. The epidemiologic studies on Pb exposure
and male fertility are limited. Previous epidemiologic studies were conducted among men seeking help at
fertility clinics. Study details of the recent epidemiologic studies are in Table 8-16. Previous and recent
toxicological evidence regarding the effect of Pb on male fertility is scarce, but generally reports reduced
fertility in males exposed to Pb. Study details of the recent toxicological studies are in Table 8-17.

8.6.3.1 Epidemiologic Studies on Male Fertility

The epidemiologic studies included in the 2013 Pb ISA that assessed associations between Pb
exposure and male fertility reported inconsistent findings. The few studies available for review were
conducted with cases that included men seeking help at fertility clinics, resulting in limited
generalizability of the studies because the study populations are not representative of the general
population. Additionally, by recruiting men who were seeking help at fertility clinics, there could be
selection bias, as their fertility status is already known and those seeking help at fertility clinics may be
different from men who have fertility issues who may be unaware of their condition and not seeking help
at a fertility clinic. Another study was conducted among occupationally exposed men, which may result in
differential exposures compared to the general population. Furthermore, another study did not control for
potential confounders.

There were a limited number of recent epidemiologic studies that examined associations between
Pb and male fertility. In the LIFE Study, a cohort of couples were followed prospectively to assess
persistent environmental chemicals and human fecundity ("Louis et aL 2012). BLLs in both female and
male partners were collected at baseline. While female blood Pb was not associated with increased time to
pregnancy, there was decreased odds, or increased time to pregnancy, for male BLLs in both the male
exposure model (OR: 0.85 [95% CI: 0.73, 0.99]) and the couple exposure model (OR: 0.82 [95% CI:
0.68, 0.97]). In a cohort of 195 couples undergoing IVF, Pb was measured in blood serum and follicular
fluid from the female partner and semen from the male partner in association with six IVF outcomes
(Zhou et aL 2021a). There was a positive association between Pb in seminal plasma and the possibility of
obtaining a good embryo (RR: 1.86 [95% CI: 1.05, 3.11]), but the associations were null across all other
IVF outcomes (normal fertilization, blastocyst formation, high-quality blastocyst, pregnancy, or live
birth).

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8.6.3.2	Toxicological Studies on Male Fertility

Only a few studies on the effects of Pb on male fertility were summarized in the 2013 Pb ISA.
These studies reported that Pb-exposed males produced smaller litters and fewer implantations and
fetuses per dam ("Anium et ah. 20.1.1; Sainath et aL 20.1.1). Only a single recent study investigated fertility
outcomes in males exposed to Pb (Table 8-17). This study exposed ICR-CD-I mice from PND 91 to 136
via drinking water and reported that sperm from treated mice had reduced fertilization capacity, resulting
in fewer fertilized oocytes in vitro (9.4 (.ig/dL) (Godinez-SoKs et aL 20.1.9).

8.6.3.3	Integrated Summary of Male Fertility

Similar to the 2013 Pb ISA, there were only a few epidemiologic studies evaluating associations
between Pb exposure and male fertility and the findings were inconsistent. The results from these studies
are limited by the small sample size and the study population was recruited from a fertility clinic, which
may have resulted in selection bias and limits generalizability as the study population has already been
diagnosed and are seeking treatment for fertility issues. Further, male fertility related to pregnancy
outcomes, such as successful implantation and normal fertilization, are more likely to be ascertained from
couples seeking treatment at fertility clinics. Additionally, different biomarkers were used to assess Pb
exposure, as well as different metrics of male fertility across the studies. In terms of toxicological
evidence, previous and recent studies are few in number. However, all report a reduction of male fertility
in Pb-treated animals using outcomes such as litter size, implantations, and fertilized oocytes.

8.6.4 Effects on Morphology and Histology of Male Sex Organs

The toxicological studies in the 2013 Pb ISA supported historical findings that showed an
association between Pb exposure and changes in the sex organs as well as germ cells. There were no
epidemiologic studies available for review for the 2013 Pb ISA that examined the relationship between Pb
exposure and morphology or histology of male sex organs. The current epidemiologic and toxicological
studies examining the relationship between Pb exposure and effects on morphology and histology of male
sex organs are summarized in the text below with study details in Table 8-16 and Table 8-17,
respectively.

8.6.4.1 Epidemiologic Studies of Morphology and Histology of Male Sex Organs

In the 2013 Pb ISA, there were no epidemiologic studies available that evaluated Pb
concentrations and associations with morphology or histology of male sex organs. A recent cohort study
evaluated the associations between prenatal metal exposure and reproductive development in boys at 2-
3 years (Huang et aL 2020). Serum concentrations of multiple metals, including Pb. were obtained from

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mothers in the Guangxi Birth Cohort Study throughout pregnancy, while reproductive development was
measured as TV and anogenital distance (AGD), categorized as anopenile distance (AGDap) and
anoscrotal distance (AGDas), in 2-3-year-old male children. When maternal serum Pb levels were
categorized by quartiles, infants in the highest quartile (serum Pb >1.23 |ig/L) had, on average, a
0.064 mL (95% CI: -0.124, -0.004) smaller TV, 0.060 cm (95% CI: -0.110, -0.011) shorter AGDap, and
0.115 cm (95% CI: -0.190, -0.039) shorter AGDas than infants in the lowest quartile (serum Pb <
0.54 |ig/L).

8.6.4.2 Toxicological Studies of Morphology and Histology of Male Sex Organs

This section is divided into the two main outcomes for the male sex organs: changes in weight of
male sex organs and changes in histology/morphology of male sex organs. The 2013 Pb ISA summarized
several studies that investigated the effects of Pb exposure on male sex organ weights. Several studies
reported decreases in weights of organs such as the testis, epididymides, vas deferens, seminal vesicles,
and prostate (Anium et ah. 20.1.1; Filial et ah. 20.1.0; Dong et ah. 2009; Salawu et ah. 2009; Biswas and
Ghosh. 2006; Rubio et ah. 2006). The direction of effect was consistent, and any effects observed were
only decreases in organ weights. However, the 2013 Pb ISA noted that there were many other studies that
did not report effects on male reproductive organ weights even when using similar doses as those studies
that did observe effects, indicating that the impact of Pb on reproductive organ weights is somewhat
inconsistent. Recent studies have also investigated the effects of Pb exposure on male sex organ weight
(Table 8-17). Wang et ah (20.1.3a) reported that dosing male CD-I mouse pups via their dams' drinking
water from PND 0 to 21 led to reduced absolute weight of testes in both treatment groups at weaning and
reduced relative testis weight in the highest treatment group at weaning (BLLs 19.1-21.2 (ig/dL on
PND 22 and 3.24-4.40 (ig/dL on PND 70). However, they observed no effect on the weight of the
prostate, seminal vesicle, or epididymides at weaning and no effects on relative weights of any
reproductive organ on PND 70. Similarly, another study reported that dosing Sprague-Dawley rats from
GD -10 to PND 183 had reduced absolute and relative testis weights (BLLs 18.6 (.ig/dL) (Wang et ah.
2013 b'). However, some studies reported that Pb did not alter the weights of testes or epididymides in ICR
mice (BLLs 6.02-21.66 ug/dL) (Pavlova et ah. 2021; Satapathv and Panda. ).

The 2013 Pb ISA reported on studies that investigated the effects of Pb on the histopathology of
male sex organs in rodents exposed to Pb. One of the most common outcomes was alterations of
seminiferous tubule pathology, such as reduced length of some spermatogenic cycle stages within
seminiferous tubules, tubule damage, and tubule atrophy (El Sfaafai et ah. 20.1.1; Shan et ah. 2009;
Massanvi et ah. 2007; Rubio et ah. 2006; Wang et ah. 2006). A few recent Pb studies have also reported
Pb-induced histopathological changes in male sex organs (Table 8-17). All recent studies were conducted
in mice, and exposure paradigms used between recent studies varied from developmental to exposure
only during adulthood. One study in CD-I mice that utilized developmental exposure (dosing dams from
lactational day 0 to 21) reported that Leydig cell numbers in the testes were reduced in the highest dose

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group at weaning and layers of spermatogenic cells within the seminiferous tubules were decreased in
both dose groups at weaning and PND 70 (BLLs at weaning 19.1-21.1 (ig/dL; BLLs at PND 70 3.24-
5.09 (.ig/dL) (Wang et ah. 2013a'). Some studies that dosed mice for 90 days following weaning reported
histopathological disruptions to the epididymal epithelial cells (BLLs 6.02-11.8 (.ig/dL) (Xie et ah. 2020)
and that spermatogenic cells within seminiferous tubules were reduced in number (BLLs at 11.92 (ig/dL)
(Zhang et ah. 2021). Lastly, a study that dosed mice from PND 60 to 74 reported that the epithelium of
the seminiferous tubules was disorganized, the luminal region contained undifferentiated germ cells, and
some tubules had decreased diameter and germ cell number and displayed incomplete spermatogenesis
(BLLs 21.7 (.ig/dL) (P avlova et ah. 2021).

A few previous and recent studies concurrently investigated the effects of Pb on male sex organ
weight and histopathology, and effects within studies were coherent (Rubio et ah. 2006; Wang et ah.

20.1.3a; Pavlova et ah. 2021). Both Rub io et ah. 2006 and Wang et ah. 20.1.3a reported reductions in male
sex organs as well as histopathological alterations, while Pavlova et ah. 202.1. reported no effects of Pb on
sex organ weight or histopathology.

8.6.4.3 Integrated Summary of Morphology and Histology of Male Sex Organs

In the 2013 Pb ISA, there were no epidemiologic studies available that evaluated Pb
concentrations and associations with morphology or histology of male sex organs. A recent cohort study
reported decreased TV, shorter AGD, shorter anopenile distance, and shorter anoscrotal distance in 2-3-
year-old male children. While it is difficult to judge coherence and consistency from the findings of a
single study, this well-designed longitudinal cohort study does provide limited evidence of changes in
morphology and histology of male sex organs. Previous and recent toxicological studies are consistent in
reporting that Pb affects different aspects of sex organ histopathology. The most consistent effects appear
to be disruptions of histopathology of seminiferous tubules within the testes. However, there exists a data
gap regarding the effects of Pb on histopathology of other male sex organs such as the prostate,
epididymides, and seminal vesicles.

8.7 Biological Plausibility

This section describes the biological pathways that may underlie some reproductive and
developmental health effects from exposure to Pb. Figure 8-1 graphically depicts the proposed pathways
as a continuum of pathophysiological responses—connected by arrows—that may ultimately lead to the
observed delayed onset in both males and females and reduced sperm/semen production, quality, and
function. This discussion of how exposure to Pb may lead to these reproductive and/or developmental
events also provides biological plausibility for the epidemiologic results reported previously in this

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Appendix. In addition, most studies cited in this subsection are discussed in greater detail earlier in this
Appendix.

Disruption of
GnRH levels

and
hypothalamic-
pituitary-gonadal

Pb
Exposure

Delayed Pubertal
Onset in Males and
Females

Damage to
organs and
supportive
somatic cells

Reduced
Sperm/Semen
Production, Quality,
and Function

GnRH = gonadotropin-releasing hormone; Pb = lead.

Note: The boxes above represent the effects for which there is experimental or epidemiologic evidence related to Pb exposure, and
the arrows indicate a proposed relationship between those effects. Solid arrows denote evidence of essentiality as provided, for
example, by an inhibitor of the pathway or a genetic knockout model used in an experimental study involving Pb exposure. Arrows
may connect individual boxes, groupings of boxes, and individual boxes within groupings of boxes. Progression of effects is
generally depicted from left to right and color-coded (gray, exposure; green, initial effect; blue, intermediate effect; orange, effect at
the population level or a key clinical effect). Here, population level effects generally reflect results of epidemiologic studies. When
there are gaps in the evidence, there are complementary gaps in the figure and the accompanying text below.

Figure 8-1 Potential biological pathways for reproductive and developmental
effects following exposure to Pb.

8.7.1 Pubertal Onset

When considering the available health evidence, plausible pathways connecting Pb exposure to
two health endpoints reported in epidemiologic and toxicological studies are proposed in Figure 8-1. The
first endpoint addressed in the figure above is delayed pubertal onset due to Pb exposure. Several
previous epidemiologic and toxicological studies that reported delays in pubertal onset in females
(Gollcnbcrg et al.. 2010; Naicker et al.. 2010; Dumitrescu et al.. 2008a; Iavicoli et al.. 2006; Denham et
al.. 2005; Selevan et al.. 2003; Wu et al.. 2003) and males (Williams et al.. 2010; Hauser et al.. 2008)
were summarized in the 2013 Pb ISA and several toxicological studies were summarized the 2006 Pb
AQCD (Pine et al.. 2006; Dearth et al.. 2004; Iavicoli et al.. 2004; Dearth et al.. 2002; Ronis et al.. 1998a.
1996). Some toxicological studies from the 2006 Pb AQCD also reported delays in pubertal onset in
males (Ronis et al.. 1998c; Sokol et al.. 1985). While no recent PECOS-relevant toxicological studies that
investigated the effects of Pb on pubertal onset were available, several recent epidemiologic studies

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reported associations between Pb exposure and delayed onset of puberty in males (Williams et ah. 20.1.9;
Nkomo et ah. 20.1.8; De Craemer et ah. 20.1.7) and females ("Gomula et ah. 2022; Jansen et ah. 20.1.8;
Nkomo et ah. 20.1.8; De Craemer et ah. 20.1.7; Slawinska et ah. 2012). The proposed biologically plausible
pathway through which Pb induces delays in pubertal onset begins with the Pb-induced disruption of the
gonadotropin-releasing hormone (GnRH) levels which may occur through reduction of circulating IGF-1
levels. GnRH is a key hormone in the hypothalamic-pituitary-gonadal axis and hormonal signaling
pathways related to reproduction and pubertal onset. A recent epidemiologic study found negative
associations between BLLs at 8-9 years of age and IGF-1 in boys 2 and 4 years later (Fleisch et ah. 20.1.3)
and toxicological studies have reported reduced IGF-1 levels and IGF-1R expression in the brains of
animals exposed to Pb (Li et ah. 20.1.6; Li et ah. 20.1.4; Dn ah. 2002; Ron is et ah. 1998b). IGF-1 is
known to act on GnRH neurons and affect GnRH secretion (Dees et ah. 2021; Daitary and Gore. 2005).
which is responsible for the release of LH and FSH from the anterior pituitary, resulting in stimulation of
the gonads to begin producing sex steroid hormones and mature oocytes and spermatozoa. One
toxicological study conducted in female Fisher 344 rats found that Pb-induced delays of pubertal onset
could be reversed by supplementation with IGF-1 (Pine et ah. 2006). This study reported that
supplementation with IGF-1 also restored GnRH and LH levels in Pb-exposed rats, demonstrating that
IGF-1 disruption is a key component in delays in the onset of puberty mediated by Pb at BLLs at/above
35 (ig/dL.

Pb has also been shown in some in vitro studies to directly alter steroidogenic enzyme expression
(e.g., steroidogenic acute regulatory protein, 3(3-hydroxysteroid dehydrogenase, and aromatase) and levels
of sex steroid hormones important for proper sexual maturation, including progesterone, E2, and
testosterone (Huang ;	2004; Srivastava et ah. 2004; Taupeau et ah. 2003; Huang et ah. 2002;

Thoreux-Manlav et ah. .1.995). Additionally, although not all studies report relationships between Pb and
hormone levels, some epidemiologic studies have reported associations and some toxicological studies
have demonstrated effects of Pb exposure on steroidogenic enzymes and sex steroid hormones (Pollack et
ah. 20.1.1; Toroouro et ah. 20.1.0; Dumitrescu et ah. 2008b; Nampoothiri and Gupta. 2008; Telisman et ah.
2007; Rubio et ah. 2006; Sokol et ah. .1.985). Pb-induced disruptions of the hypothalamic-pituitary-
gonadal axis, steroidogenic enzymes, and their sex steroid products are plausible explanations for the
observed delays in pubertal onset reported in epidemiologic and toxicological studies.

8.7.2 Male Reproductive Function

The other health outcome proposed in Figure 8-1 is male reproductive function. Recent
epidemiologic studies have reported that Pb exposure is associated with reductions in a variety of semen
parameters, including sperm motility, sperm concentration, and normal sperm morphology (Shi et ah.
202.1.; Sukhn et ah. 20.1.8; Li et ah. 20.1.5; Pant et ah. 20.1.4). These findings are generally consistent with
the epidemiologic evidence presented in the 2013 Pb ISA (U.S. EPA. 20.1.3). Further, toxicological studies
provide supporting evidence that Pb negatively impacts male reproductive function (see Section 8.6.1)

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(Aniiiro et ah. 20.1.1; Sainath etah. 20.1.1). Figure 8-1 shows a plausible biological pathway through which
Pb may act to reduce reproductive function in males.

The 2013 Pb ISA concluded that the evidence indicates a causal relationship between Pb
exposure and reduced quality of sperm, and that this relationship was likely mediated through the
generation of reactive oxygen species (ROS). leading to cellular damage ("U.S. EPA. 2013). Specifically,
the 2013 Pb ISA summarized one study that reported Pb-induced increases in oxidative stress markers
and reductions in antioxidant enzyme levels in testicular plasma of rats ("Salawu et ah. 2009). In addition,
several studies in the 2013 Pb ISA reported attenuation of Pb-induced reductions in sperm count, motility,
and viability when animals were co-administered substances with known antioxidant properties ("Salawu
et ah. 2009; Shan et ah. 2009; Madhavi et ah. 2007; Rubio et ah. 2006; Wang et ah. 2006). Further
supporting the proposed pathway through which oxidative stress mediates Pb-induced effects are
additional studies that report that Pb exposure dysregulates antioxidant enzymes, leading to oxidative
stress and DNA damage in the affected tissues (Lopes et ah. 20.1.6; Kagi and Vallee. .1.960; Ommati et ah.
In Press). Recent studies also support the proposed pathway and report an attenuation of Pb-induced
effects on aspects of male reproductive function (e.g., subfecundity, reduced sperm count) in animals
supplemented with antioxidants (Zhang et ah. 2021; Abdelhamid et ah. 2020; Alotaibi et ah. 2020; Naderi
et ah. 2020; Udefa et ah. 2020; Abdrabou et ah. 20.1.9; Hassan et ah. 20.1.9; Ommati et ah. 20.1.9;
BaSalamah et ah. 20.1.8; Hasanein et ah. 20.1.8; Mabrouk. 20.1.8; El Shafai etah. 20.1.1; Leiva et ah. 20.1.1;
Sainath etah. 20.1.1; Ommati et ah. In Press). Although many studies report negative effects of Pb on
supporting somatic cells that have key functions in the spermatogenic cycle (e.g., Leydig cells, Sertoli
cells), Pb may also have negative effects directly on sperm cells. Direct contact of Pb with sperm cells has
been documented by multiple studies (Jia et ah. 2022; Sukhn et ah. 20.1.8; Pant et ah. 20.1.4). One recent
study reported that incubating sperm from healthy adult men for 4 hours with 30 |ig/m L or 8 hours with
either 15 or 30 |ig/mL Pb increased DNA fragmentation, possibly due to oxidative stress and Pb binding
to DNA phosphate residues, disrupting the process of chromatin condensation (Gomes et ah. 20.1.5). In
another study, 4 hours of incubation of semen samples from healthy adult men with Pb reduced
intracellular levels of cyclic adenosine monophosphate (cAMP) (10, 50, and 100 |iM) and Ca2+ (2.5, 10,
50, and 100 (j,M), both of which are important in regulating sperm cell function (He et ah. 20.1.6). In
support of this alternative mechanism of action is one non-PECOS relevant study (due to use of i.p.
injection route) that reported an attenuation of Pb-induced effects on reproduction in Pb-injected male
mice that were supplemented with CaCl; (Golshan Iran pour and Kfaeiri. 20.1.6). Disruption of intracellular
levels of key components such as cAMP and Ca2+is another way in which Pb can directly affect sperm
health and function outside of oxidative stress.

In summary, pathways are suggested by which Pb exposure can delay pubertal onset and reduce
sperm/semen production, quality, and function. Studies indicate that Pb exposure likely impacts the
hypothalamic-pituitary-gonadal axis in both males and females, leading to disruption of the onset of
puberty, a developmental period with increasing regard for its sensitivity to insult due to the vulnerability
of the various endocrinological events for which it is known. In addition, Pb exposure alters multiple

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aspects of male reproductive function. The production of adequate quantities of viable sperm is essential
for proper male fertility and reproduction. Pb exposure hampers this by negatively impacting both the
sperm cell and the supportive somatic cells that play key roles in the spermatogenic cycle through
increased oxidative stress and disruption of other important intracellular functions.

8.8 Summary and Causality Determination

The 2013 Pb ISA (U.S. EPA. 20.1.3') made four causality determinations for Pb exposure and
(1) effects on pregnancy and birth outcomes; (2) effects on development; (3) effects on female
reproductive function; and (4) effects on male reproductive function. The 2013 Pb ISA concluded that the
evidence is suggestive of a causal relationship between Pb exposure and effects on birth outcomes; a
causal relationship between Pb exposure and effects on development, based on the findings of delayed
pubertal onset among males and females; suggestive of a causal relationship between Pb exposure and
effects on female reproductive function; and a causal relationship between Pb exposure and effects on
male reproductive function. The following sections detail the causality determinations based on the recent
epidemiologic and toxicological studies.

8.8.1 Summary of Effects on Pregnancy and Birth Outcomes

The 2013 Pb ISA concluded that based on the mix of inconsistent results of studies on various
birth outcomes and some associations observed in epidemiologic studies of preterm birth and low birth
weight/fetal growth, the evidence was suggestive of a causal relationship between Pb exposure and birth
outcomes. Some associations were observed between Pb and low birth weight in epidemiologic studies
that used postpartum maternal bone Pb or air Pb concentrations. Although associations were less
consistent for low birth weight with maternal blood Pb measured, during pregnancy or at delivery, or with
Pb measured in the umbilical cord and placenta (maternal blood Pb or umbilical cord and placenta Pb
were the biomarkers most commonly used in studies of low birth weight), some negative associations
between Pb biomarker levels and low birth weight or other measures of fetal growth were observed. The
effects of Pb exposure during gestation in animal toxicological studies included mixed findings, but most
studies reported reductions in birth weight of pups or birth weight of litters when dams were treated with
Pb. Thus, although evidence available was mixed, some associations observed in epidemiologic studies of
preterm birth and low birth weight or fetal growth provided suggestive evidence of a causal relationship
between Pb exposure and birth outcomes.

Compared to the evidence assessed in the 2013 Pb ISA, the evidence for associations between Pb
exposure and birth outcomes in the 2024 Pb ISA is notably stronger due to a recent quasi-experimental
study demonstrating decreased probability of preterm birth, decreased probability of low birth weight,
decreased probability of SGA, and increased birth weight (Bui et al.. 2022). Overall, among the recent

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epidemiologic studies, there was a pattern of elevated risk of preterm birth observed across several studies
from multiple geographic locations. Additionally, the recent epidemiologic studies of preterm birth
included populations for which mean/median maternal blood Pb values were below 10 (ig/dL and
controlled for wide range of confounders, including GA, other metals, and maternal health factors
(e.g., smoking, parity, BMI). There remain uncertainties regarding the critical window for the timing of
the exposure (e.g., during pregnancy, at delivery), biomarkers examined for Pb (e.g., maternal blood, cord
blood, maternal red blood cells, maternal serum, placental tissue), and evaluation of co-pollutants among
the epidemiologic literature with limited supportive evidence in the toxicological literature. While there
were no epidemiologic or toxicological studies examining Pb exposure and maternal health outcomes in
the 2013 Pb ISA, recent epidemiologic and toxicological studies reported inconsistent results regarding
maternal health outcomes and different maternal health outcomes were evaluated between the
epidemiologic and toxicological studies. Among the epidemiologic studies, there were consistent null
associations between maternal blood (blood, serum, and erythrocytes) Pb levels and GDM in studies that
reported mean/median blood Pb below 10 (ig/dL. Although some recent epidemiologic studies
investigated various pregnancy-related endpoints, the small number of studies limits the ability to judge
coherence and consistency across these studies. Among the few toxicological studies that investigated
maternal health, the only outcome reported was maternal weight gain during pregnancy. Most studies
reported no effects of Pb on maternal weight gain during pregnancy, and the few that reported reductions
in maternal weight gain also reported reductions in dam brain weight, a marker often indicative of overt
toxicity. This suggests that the observed reduction in maternal weight gain during pregnancy reported in
these studies may not be directly due to Pb exposure and may have been influenced by overt toxicity
experienced by the dams.

The recent epidemiologic and toxicological studies of birth outcomes reported inconsistent
findings overall. Among the recent epidemiologic studies of prenatal growth and Pb exposure, the
findings were inconsistent, and no effects on birth weight were reported in the recent toxicological
studies. The inconsistencies in the recent epidemiologic studies of prenatal growth and Pb exposure may
be due to differences in study design, the timing of the exposure, differences in biomarkers of exposure,
and the wide variation in prenatal growth outcomes assessed (birth weight, birth length, HC, GA). A few
studies were further limited by small sample size, which may cause imprecision in the measures of
association. Recent toxicological studies did not report any effects of Pb exposure on birth weight. Of
note is a previous study discussed in the 2013 Pb ISA that reported reduced litter weights at birth were
driven by reduced weights in female pups. No recent studies performed separate analyses of birth weight
for male and female pups, or they did not assess female pup weights at all. This suggests that the observed
lack of effects in recent literature could be due to a lack of sensitivity.

The recent epidemiologic studies of Pb exposure and birth defects, specifically NTDs, CHDs,
OFC defects and abdominal congenital malformations, reported inconsistent associations. While the
associations were generally null for Pb exposure (measured in placental tissue, umbilical tissue, maternal
blood serum, and umbilical cord serum) and NTDs, CHDs, and abdominal congenital malformations,

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there were positive associations with OFC defects when Pb was measured in placental tissue or maternal
blood. The small number of studies limits the ability to judge consistency and coherence across studies of
different birth defects (e.g., NTDs, CHDs, OFC defects, and abdominal congenital malformations), timing
of Pb exposure (e.g., second trimester, third trimester, and at delivery), differences in biomarkers
(e.g., placental tissue, umbilical tissue, maternal blood serum, and umbilical cord serum, maternal blood),
and confounders considered in the analyses. Additionally, the relatively small sample sizes in some
studies reduce the statistical power to determine the precision of the associations. Recent toxicological
studies report no effects of Pb on birth defects in offspring. This contrasts with some previously reviewed
studies that reported defects in offspring of Pb-exposed dams. However, dams in these previous studies
also experienced overt toxicity due to the high Pb doses used, which did not occur in recent toxicological
studies, suggesting that maternal toxicity may have been involved with the birth defects observed in
previous studies.

There were only a few recent epidemiologic studies that evaluated Pb exposure and spontaneous
abortion and pregnancy loss. Studies that examine spontaneous abortion are difficult to conduct as many
spontaneous abortions or pregnancy losses occur during the first trimester. Women may miscarry before
being enrolled in a study and/or women may not have known they were pregnant when they miscarried,
thus limiting the ability of a study to detect subtle effects (e.g., if higher Pb exposures lead to increased
risk of early spontaneous abortions). In the recent epidemiologic studies, some of the studies assessing
spontaneous abortion and/or pregnancy loss were among women who were undergoing treatment at
fertility clinics. Detection of spontaneous abortion and/or pregnancy loss is more likely to be ascertained
in such clinics, but this study design approach may result in selection bias and limited generalizability of
the results because the study populations are not representative of the general population as they have
already been diagnosed and are seeking treatment for infertility. In the recent toxicological studies, there
were no reported effects of Pb exposure on pre- or postnatal offspring mortality. Although not always
consistently so, BLLs were generally lower in recent toxicological literature when compared to previous
literature, possibly explaining the observed contrast in results.

There were no epidemiologic studies available that evaluated Pb concentrations and associations
with placental function in the 2013 Pb ISA. There were a limited number of recent epidemiologic studies
in this area. These cross-sectional studies provide insight into associations between concurrent Pb
exposure and placental function, but are limited by their cross-sectional design, making it difficult to
establish the temporality of the effects or the critical window of exposure to Pb that might result in
changes in the placenta during pregnancy. Further, there were only a small number of cases, which may
result in imprecise associations. While previous toxicological evidence included decreased placental
weight and histological alterations, these findings were limited to a single study. Recent toxicological
studies reported that dams dosed with Pb had reduced placental weight, but some of these studies also
reported reduced brain weight in dams, suggesting that overt toxicity may have occurred and could be
related to the observed reductions in placental weight.

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There were also a number of recent epidemiologic studies that evaluated other outcomes related
to maternal health during pregnancy such as biomarkers of fetal immune system, fetal marker for
metabolic function, and rTL, but the small number limits the ability to judge the coherence and
consistency across these studies. The only additional pregnancy outcome investigated in recent
toxicological literature was sex ratio of offspring born to Pb-treated dams. Although most toxicological
studies reported no effects of Pb on sex ratio, a single study reported that Pb produced female-skewed
litters when compared to control. It is worth noting, however, that the non-Pb-exposed groups were male-
skewed and had male:female offspring ratios of 1.4-1.5, whereas Pb-treated groups had male:female
ratios of 1 to 1.

In summary, the collective evidence is sufficient to conclude that there is likely to be a causal
relationship between Pb exposure and effects on pregnancy and birth outcomes. This determination
is largely driven by a recent quasi-experimental study that reported Pb-related changes in birth weight and
probability of low birth weight, preterm birth, and small for gestational age, in addition to other studies
demonstrating effects between Pb exposure and preterm birth (Table 8-1). Additionally, there were a few
high-quality epidemiologic studies that reported associations with relevant BLLs and prenatal growth,
birth defects, spontaneous abortion and pregnancy loss, and placental function, but the findings overall
were inconsistent. There is uncertainty related to exposure patterns resulting in likely higher past Pb
exposures, especially among maternal Pb levels. Additional uncertainties are related to biomarkers of
exposure (maternal blood, maternal serum, maternal bone, maternal erythrocytes, cord blood, cord blood
serum, placental tissue), the critical window of exposure, and co-pollutants confounding. Of note, the
cohorts in the recent epidemiologic literature would generally be expected to have had appreciable past
exposures to Pb; however, the extent to which adult BLLs in these cohorts reflect the higher exposure
histories is unknown as is the extent to which these past Pb exposures (magnitude, duration, frequency)
may or may not elicit effects on pregnancy and birth outcomes. The recent evidence from the
toxicological studies mostly reported no effects of Pb across pregnancy and birth outcomes. This may be
due to the exclusion of toxicological studies with BLLs greater than 30 (ig/dL, indicating the possibility
that most pregnancy and birth outcomes are only affected in laboratory animals at levels higher than most
environmentally relevant Pb exposure levels.

8.8.2 Summary of Effects on Development

The 2013 Pb ISA concluded that the collective body of evidence integrated across epidemiologic
and toxicological studies, based on the findings of delayed pubertal onset among males and females, was
sufficient to conclude that there is a causal relationship between Pb exposure and developmental effects.
Multiple epidemiologic studies of Pb and puberty in the 2013 Pb ISA showed associations between
concurrent BLLs and delayed pubertal onset for girls and boys. In cross-sectional epidemiologic studies
of girls (ages 6-18 years) with mean and/or median concurrent BLLs from 1.2 to 9.5 (ig/dL, consistent
associations with delayed pubertal onset (measured by age at menarche, pubic hair development, and

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breast development) were observed. In boys (ages 8-15 years), fewer epidemiologic studies were
conducted but associations between BLLs and delayed puberty were observed, including associations
among boys in a longitudinal study. These associations were consistently observed in populations with
mean or median BLLs of 3.0 to 9.5 (ig/dL. Potential confounders considered in the epidemiologic studies
of both boys and girls that performed regression analyses varied. Most studies controlled for age and
BMI. Other variables, such as measures of diet, socioeconomic status (SES), and race/ethnicity, were
included in some of the studies. Adjustment for nutritional factors was done less often and this could be
an important confounder. A study using a cohort of girls from NHANES controlled for various dietary
factors as well as other potential confounders and reported an association between increased concurrent
BLLs and delayed pubertal onset ("Selevan et aL 2003). A limitation across most of the epidemiologic
studies of BLLs and delayed puberty was the cross-sectional design, which does not allow for an
understanding of temporality. There was uncertainty with regard to the exposure frequency, timing,
duration, and level that contributed to the associations observed in these studies. Additionally, the
toxicological studies reviewed in the 2013 Pb ISA indicated that delayed pubertal onset may be one of the
more sensitive developmental effects of Pb exposure with effects observed after gestational exposures
leading to BLLs in the female pup of 1.3-13 (ig/dL (lavieoli et aL 2006; lavicoli et aL 2004). An
additional study reviewed in the 2013 Pb ISA reported increases in age at vaginal opening in Wistar rats
that were dosed prior to conception and in utero, but BLLs were not reported (Dumitrescu et aL 20083).
These results are supported by studies reviewed in the 2006 Pb AQCD that reported delays in pubertal
onset in female rats and mice as measured by age at vaginal opening and age at first estrus ("Pine et aL
2006; Dearth et aL 2004; Dearth et aL. 2002; Ron is et aL 1998a; Ron is et aL 1998c; Ron is et aL .1.996).
BLL varied greatly between studies with some reporting effects occurring in dose groups with levels
below 30 (ig/dL (Dearth et aL 2004; Dearth et aL. 2002). while others only report effects in groups with
BLLs higher than 30 (ig/dL (Ron is et aL. 1998a; Ronis et aL. 1998c; Ron is et aL. 1996V A key study
reviewed in the 2006 Pb AQCD, Pine et al. (2006) reported increased age at vaginal opening in Fisher
344 rats that was attenuated by supplementation of IGF-1. However, Pine et al. (2006) only reported
BLLs of dams (39.8 (.ig/dL). making it difficult to determine what BLLs in the offspring may have been
achieved to elicit such effects on puberty. Toxicological studies have also reported delayed male sexual
maturity as measured by sex organ weight, among other outcomes, seeing significant decrements at BLLs
of 20-34 (ig/dL (Ronis et al.. 1998c; Sokol et al.. .1.985). Thus, the 2013 Pb ISA concluded that the data
from the toxicological literature and from epidemiologic studies demonstrated puberty onset in both
males and females was delayed with Pb exposure.

In the 2013 Pb ISA, findings from epidemiologic studies of the effect of Pb on postnatal growth
were inconsistent. Findings from the toxicological literature of the effect of Pb exposure on postnatal
growth summarized in the 2013 Pb ISA and the 2006 Pb AQCD were fairly consistent, and most studies
showed decreases in body weight of Pb-exposed offspring at postnatal time points, while one study
reported an increase in body weight at 1 year of age in male offspring only.

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The 2013 Pb ISA summarized some toxicological evidence that demonstrated the effect of Pb on
other developmental outcomes, including impairment of retinal development, effects on the lens of the
eye, and alterations in the developing hematopoietic, hepatic systems and teeth. No studies that
investigated more classic toxicological developmental milestones (e.g., eye slit formation, eye opening,
pinna detachment) were reported in the 2013 Pb ISA.

In the recent epidemiologic and toxicological literature, the relationships between Pb exposure
and puberty onset in both females and males, as well as postnatal growth, were reviewed. While there
were no recent PECOS-relevant toxicological studies in puberty in either females or males, the recent
epidemiologic studies reported consistent patterns of association between blood Pb exposure and delayed
age of menarche ("Gomula et ah. 2022; Jansen et ah. 20.1.8; De Craemer et ah. 20.1.7; Slawinska et ah.
20.1.2) and some indication of slower breast development (Nkomo et ah. 20.1.8; De Craemer et ah. 20.1.7) in
females, which is similar to the findings from the epidemiologic studies reviewed in the 2013 Pb ISA.
However, the associations between Pb exposure and male pubertal onset were inconsistent among the
cross-sectional studies. The differences in markers of puberty in males (hormone levels, pubic hair
development, genital development, TV) may explain the inconsistencies in findings across recent studies.
While the studies assessing Pb exposure and female and male puberty were limited by differences in the
timing of exposure to Pb or Pb biomarker (blood, maternal bone, cord blood), these studies consider a
wide range of confounders, including height, weight, and BMI.

The recent toxicological and epidemiologic studies that evaluated the relationship between Pb
exposure and postnatal growth were inconsistent. The majority of recent toxicological studies did not
report changes in postnatal growth due to Pb exposure (Zhao et ah. 2021; Xie et ah. 2020; Rao Barkur
and Bairv. 20.1.6; Basha and Reddv. 20.1.5; Basgen and Sobin. 2014). However, some recent toxicological
studies reported decreases (Duan et ah. 20.1.7; de Figueiredo et ah. 20.1.4; Graham et ah. 20.1.1) and
increases (Betharia and Maher. 2012) in body weight of offspring due to Pb exposure. Among
epidemiologic studies that evaluated the associations between blood Pb and postnatal growth in children
(older than 4 years) there were more consistent patterns of associations of decreased height and weight
(Signes-Pastor et ah. 2021; Kuang et ah. 2020; Zhou et ah. 2020; Deierlein et ah. 20.1.9; Kerr et ah. 20.1.9;
Choi et ah. 2017). Overall, there were negative associations between Pb exposure and specific postnatal
growth outcomes among the cross-sectional studies. However, among cohort studies, there were some
inconsistencies in the associations of Pb exposure and different postnatal growth outcomes. These
inconsistencies in the cohort studies may be due to differences in the timing of when Pb exposure was
measured, the biomarker of Pb exposure (maternal blood, maternal bone, cord blood, infant blood,
childhood blood), and the timing of the outcome. The current inconsistent findings of exposure to Pb and
postnatal growth are similar to those reported in the 2013 Pb ISA.

There was a small body of epidemiologic studies across various other developmental effects;
however, the small number of studies limits the ability to judge coherence and consistency across these
studies, although the associations reported demonstrate that Pb exposure could result in physiological

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responses that contribute to adverse developmental effects, including changes to diurnal Cortisol rhythms,
lower salivary sialic acid levels, and oxidative stress damage to DNA from Pb exposure among children
during developmental periods. Recent studies that investigate other developmental outcomes such as
developmental milestones are scarce. Some toxicological studies investigated developmental milestones
in rodents (e.g., pinna detachment, eye slit formation, eye opening, tooth eruption, and fur development),
but no effects of Pb exposure were reported on any of these milestones in groups with PECOS-relevant
BLLs.

In summary, the collective evidence is sufficient to conclude a causal relationship exists
between Pb exposure and effects on development. The key evidence is outlined in Table 8-1. While
there were no recent PECOS-relevant toxicological studies that investigated the impacts of Pb on puberty
in either females or males, previous toxicological evidence demonstrated that Pb exposure consistently
delayed pubertal onset in female rodents. Of note is one key previous toxicological study (Pine et ah,
2006) which demonstrated that delayed pubertal onset in female rats developmentally exposed to Pb
could be completely attenuated by supplementation with IGF-1. Further, the recent epidemiologic studies
reported consistent patterns of associations between blood Pb exposure and delayed age of menarche and
some indication of slower breast development in females, which is similar to the findings from the
epidemiologic studies reviewed in the 2013 Pb ISA. The few recent cohort studies of male pubertal onset
found consistent associations between Pb exposure and delayed onset. Recent cross-sectional studies
reported inconsistent results, possibly due to differences in the markers of puberty examined (hormone
levels, pubic hair development, genital development, TV). Though the effects of Pb exposure on postnatal
growth were inconsistent overall, there was some evidence from toxicological studies indicating reduced
body weight of offspring and from epidemiologic studies reporting associations between blood Pb and
decreased height and weight in children. The cohorts in the recent epidemiologic literature would
generally be expected to have had appreciable past exposures to Pb; however, the extent to which adult
BLLs in these cohorts reflect the higher exposure histories is unknown, as is the extent to which these
past Pb exposures (magnitude, duration, frequency) may or may not elicit developmental effects such as
decreased postnatal growth or disrupted puberty. Toxicological evidence supports biologically plausible
pathways of how Pb exposure exerts its effects on pubertal onset (Li et ah, 20.1.6; Li et ah, 20.1.4; Pine et
ah, 2006; Dearth et ah, 2002; Ron is et ah, 1998b), including studies suggesting that Pb may impact
pubertal onset via dysregulation of IGF-1 resulting in a cascade of effects that alters levels of hormones
important during the pubertal period.

8.8.3 Summary of Effects on Female Reproductive Function

The 2013 Pb ISA concluded that the available evidence was suggestive of a causal relationship
between Pb exposure and female reproductive function. Epidemiologic and toxicological studies of
reproductive function among females investigated whether Pb biomarker levels were associated with
hormone levels, fertility, menstrual/estrous cycle changes, and altered morphology or histology of female

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reproductive organs. Two previous toxicological studies conducted in non-human primates reported
disrupted menstrual cyclicity and reduced progesterone, although another non-human primate study with
lower BLLs than the other studies (<40 (ig/dL versus 44-89 (ig/dL) reported no effects on menstrual
cyclicity. Some previous toxicological studies in rodents also demonstrated impacts of Pb exposure on
estrous cyclicity, but of note is the high BLLs in treated animals (63.2-264 (ig/dL). Some of the
epidemiologic studies reviewed in the 2013 Pb ISA reported associations with concurrent BLLs and
altered hormone levels in adults, but results varied among studies, possibly due to the different hormones
examined and the different timing in menstrual and lifecycles. There was some evidence of a potential
relationship between Pb exposure and female fertility, but findings were mixed. The majority of the
epidemiologic studies were cross-sectional and adjustment for potential confounders varied from study to
study, with some potentially important confounders, such as BMI, not included in all studies. Further,
most of the epidemiologic studies on female reproductive function reviewed in the 2013 Pb ISA had
small sample sizes and were generally conducted in women attending infertility clinics. Previous
toxicological studies reported inconsistent effects of Pb on female fertility outcomes in rodents (reduced
litter size, reduced number of litters produced), while all recent toxicological studies reported no effects
of Pb exposure on female fertility outcomes. Studies that reported impacts of Pb on female fertility
outcomes tended to use higher doses, longer dosing durations, and/or concurrently exposed sires in
addition to dams, which may explain the observed contrast between studies. Although epidemiologic and
toxicological studies provide information on different exposure periods, both types of studies, including
some high-quality epidemiologic and toxicological studies, supported the conclusion that Pb may affect
some aspects of female reproductive function.

There were no recent PECOS-relevant toxicological studies of the effects of Pb exposure on
hormone levels in females or menstrual/estrous cyclicity; however, there were several recent
epidemiologic studies. The recent epidemiologic studies examining the relationship between Pb exposure
and hormone levels reported consistent positive associations between blood Pb and FSH and LH in
women who were post-menopausal. While these studies were limited by their cross-sectional study
design, the studies were conducted in well-established population-based surveys. These studies
considered a range of confounders, including controlling for BMI, smoking, and co-exposure with Cd, but
not all studies adjusted for some potential important confounders such as age at menarche, pregnancy
history, oral contraceptive use, and female hormone use, such as IVF or hormone therapy. Additionally,
the recent studies examining the relationship between menopause and Pb exposure found consistent
positive associations of early risk of menopause. The results from a study of concurrent exposure of blood
Pb with menopause were supported by the results from a longitudinal cohort that reported that bone Pb, a
cumulative biomarker of Pb exposure, was associated with difference in age at menopause and risk of
early menopause.

Among the recent epidemiologic studies, there were inconsistent associations between Pb
exposure and female fertility. In studies among participants in the general population, there was an
increased risk of self-reported infertility and longer time to pregnancy (Lee et ah. 2020; Louis et al..

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2012). However, among studies with women who were either seeking help at a fertility clinic or reported
infertility the associations were inconsistent. Because the study participants included only a small sample
of women who were either seeking help at a fertility clinic or self-reported infertility, selection bias may
exist and limits the generalizability of the results. Additionally, these studies were limited by the
concurrently measured exposure and outcome, different biomarkers of exposure (blood, serum, and
follicular fluid), and a small number of participants. These studies did include adjustment for potential
confounders, including age, BMI, and partner exposure. The recent toxicological studies in female
fertility did not observe alterations in the number of litters or the litter size in Pb-exposed dams that began
dosing prior to conception.

There was only a single recent epidemiologic study evaluating the association between Pb
exposure and morphology or histology of female sex organs (ovaries, uterus, fallopian tubes/oviducts,
cervix, vagina, and/or mammary glands) and no recent PECOS-relevant toxicological studies. The results
from the single epidemiologic study reported null associations between blood Pb and rate of uterine
fibroids and uterine fibroid volume, but women with uterine fibroids had higher geometric mean BLLs
than women without fibroids (1.43 (ig/dL versus 1.35 (ig/dL, respectively).

In summary, the collective body of evidence is sufficient to conclude that there is likely to be
a causal relationship between Pb exposure and female reproductive function. The strongest line of
evidence is from recent epidemiologic studies examining the relationship between Pb exposure and
effects on hormone levels and menstrual/estrous cyclicity (Table 8-1). Positive associations from a
longitudinal cohort between bone Pb, a biomarker of cumulative Pb exposure, and both earlier age at
menopause and risk of early menopause were supported by results from a cross-sectional NHANES study
of concurrent exposure of blood Pb with earlier age at menopause. Additionally, recent epidemiologic
studies found consistent positive associations between blood Pb and FSH and LH in women who were
post-menopausal. While these studies are limited by their cross-sectional study design, the studies were
conducted in well-established population-based surveys. These studies considered a range of confounders,
even co-exposure to other metals, but not all studies adjusted for some potential important confounders
such as age at menarche, pregnancy history, oral contraceptive use, and female hormone use, such as IVF
or hormone therapy. While there were no recent PECOS-relevant toxicological studies that examined the
effects of Pb on hormone levels in females or menstrual or estrous cyclicity, previous toxicological
evidence suggests that Pb may disrupt reproductive hormones and menstrual and estrous cyclicity in
females. Two toxicological studies in rats reported disruptions in estrous cyclicity, and two toxicological
studies based in non-human primates reported alterations to different menstrual cycle aspects (e.g., length
of cycle, length of menstruation) and reproductive hormone levels. Additional rodent studies reported
effects of Pb on circulating reproductive hormone levels, including sex steroid hormones (progesterone,
testosterone, and E2) and gonadotropin hormones (LH and FSH).

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8.8.4

Summary of Effects on Male Reproductive Function

The 2013 Pb ISA concluded that there was sufficient evidence to support a causal relationship
between Pb exposures and male reproductive function. This determination was based on toxicological
evidence of sperm/semen production, quality, and function with supporting evidence in the epidemiologic
studies, in addition to evidence supportive for a mode of action. Previous toxicological studies with
relevant Pb exposure routes reported effects on rodent sperm quality and sperm production rate (BLL
range: 34-37 (ig/dL) ("Sokol and Berman. .1.991; Sokol et ah. .1.985). sperm DNA damage (BLL of 19 and
22 (ig/dL) (Nava-Hernandez et ah. 2009). and histological or ultrastructural damage to the male
reproductive organs in studies from rodents (BLL of 5.1 (ig/dL) (El Sfaafai et ah. 20.1.1) and non-human
primates (BLL of 43 (ig/dL) (Cullen et ah. .1.993). These effects were found in animals exposed to Pb
during peripuberty or adulthood for 1 week to 3 months. The toxicological studies reported that Pb
exposure decreased reproductive organ weight and caused histological changes in the testes and germ
cells. Subfecundity (decreased number of pups born/litter) was reported in unexposed females mated to
Pb-exposed males. Also, sperm from Pb-exposed rats (BLLs: 33 to 46 (ig/dL) used for IVF of eggs
harvested from unexposed females yielded lower rates of fertilization (Sokol et ah. .1.994). Supporting
evidence was provided by decrements in sperm quality from rabbits administered Pb subcutaneously
(BLLs of 25 (ig/dL) (Moorman et ah. .1.998).

The 2013 Pb ISA reported detrimental effects of Pb on sperm observed in epidemiologic studies
with concurrent BLLs of 25 (ig/dL and greater among men occupationally exposed (Hsu et ah. 2009;
Kasperczvk et ah. 2008; .Naha and Manna. 2007; .Naha and Cfaowdfaurv. 2006). Findings of these
epidemiologic studies are limited due to these high exposure levels among the occupational cohorts and
the lack of consideration for potential confounding factors, including occupational exposures other than
Pb. Studies among men with lower Pb levels were limited to infertility clinic studies, which may produce
a biased sample and findings that lack generalizability. However, a well-conducted epidemiologic study
that enrolled men going to a clinic for either infertility issues or to make a semen donation and controlled
for other metals and smoking reported a positive association of blood Pb with various detrimental effects
in sperm (Telisman et ah. 2007). The median concurrent BLL in this study was 4.92 (ig/dL (range: 1.13-
14.91). A similar study also reported possible associations between concurrent blood Pb and various
semen parameters, but the results were extremely imprecise (large confidence intervals [CIs]), making it
difficult to draw conclusions (Meeker et ah. 2008).

The epidemiologic and toxicological studies in the 2013 Pb ISA reported inconsistent results
regarding hormone aberrations associated with Pb exposure. Due to the complexity of the reproductive
system, uncertainty exists as to whether Pb exerts its toxic effects on the reproductive system by affecting
the responsiveness of the hypothalamic-pituitary-gonadal axis by suppressing circulating hormone levels
or by some other pathway. Inconsistent findings were also apparent among epidemiologic studies of
fertility among men.

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Toxicological studies from the 2013 Pb ISA suggested that oxidative stress was a major
contributor to the effects of Pb exposure on the male reproductive system, providing mode of action
support. The effects of ROS may involve interference with cellular defense systems leading to increased
lipid peroxidation and free radical attack on lipids, proteins, and DNA. Several studies showed that Pb
induced germ cell injury (as evidenced by aberrant germ cell structure and function) and increased
generation of ROS within the male sex organs. Co-administration of Pb with various antioxidant
compounds either eliminated Pb-induced injury or greatly attenuated its effects. In addition, many studies
that observed increased oxidative stress also observed increased apoptosis, which is likely a critical
underlying mechanism in Pb-induced germ cell dysfunction.

Recent epidemiologic and toxicological studies examined Pb exposure and male reproductive
function, including sperm/semen production, quality, and function; hormone levels; fertility; and
morphology and histology of male sex organs. Among the studies that evaluated the relationship of Pb
exposure and sperm/semen production and quality, there was consistent evidence of effects when the
exposure metric was blood Pb. In the recent epidemiologic studies, there were consistent associations of
decreased sperm/semen production and quality with increased blood Pb, but there were inconsistent
associations when Pb was measured in seminal fluid or seminal plasma. The majority of the
epidemiologic studies that evaluated the associations of Pb and sperm/semen production and quality were
cross-sectional studies conducted in males attending fertility clinics, limiting the generalizability of the
results. The studies were further limited by concurrent measurement of exposure and outcome, different
biomarkers of Pb, different seminal parameters, exposure circumstances (historical exposure, magnitude,
duration, timing, and frequency), and small sample sizes. Despite these limitations, it is important to note
that a wide variety of potential confounders were considered, including controlling for hormone levels.
The recent toxicological studies support the findings from the epidemiologic studies. Among the recent
toxicological studies, the majority reported that Pb exposure negatively impacted sperm/semen production
and quality, although these studies were limited to a single species, and no recent toxicological studies
reported on the effects of Pb on sperm or semen parameters in any other laboratory animal species.

There were a limited number of recent epidemiologic and toxicological studies that examined the
relationship between Pb and hormones in males. While recent epidemiologic and toxicological studies
reported changes in hormone levels among males, the direction of the observed relationships differed
across disciplines. Specifically, the epidemiologic studies reported Pb-associated increases in
testosterone, whereas the toxicological studies reported a reduction in testosterone following exposure to
Pb. Additionally, recent cross-sectional epidemiologic studies reported inconsistent associations between
blood Pb and other sex hormones. One study reported positive associations between blood Pb and SHBG,
FSH, and LH, as well as positive trends among quartiles of blood Pb, suggestive of a potential exposure-
response relationship. In contrast, an NHANES study reported null associations between blood Pb and
E2, fE2, androstenedione glucuronide, and SHBG. Recent toxicological evidence regarding the effects of
Pb on male sex hormones is limited to a single study that reported that exposure through the dam's milk

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from birth to weaning (PND 21) in CD-I mice was sufficient to reduce testosterone in the serum at
weaning and on PND 70.

The recent epidemiologic and toxicological studies of Pb exposure and male fertility were
limited. In the recent epidemiologic studies, male fertility was measured by IVF outcomes. There were
inconsistent associations with Pb exposure and male fertility, with one study reporting blood Pb was
associated with longer time to pregnancy, but another reported a positive association between Pb in
seminal plasma and the possibility of obtaining a viable embryo. Differences in Pb biomarkers and
difference in outcomes might explain the inconsistencies of the associations among these studies. The
males in these studies were also recruited from fertility clinics, which might have resulted in selection
bias and limits the generalizability of the results. A single recent toxicological study reported that sperm
from Pb-exposed mice had reduced fertilization capacity, resulting in fewer fertilized oocytes in vitro.

There were a limited number of studies of Pb exposure and morphology or histology of male sex
organs. There was only a single recent epidemiologic study that reported decreased TV, shorter anopenile
distance, and shorter anoscrotal distance with maternal serum Pb exposure. Among the recent
toxicological studies, Pb exposure resulted in effects in the morphology or histology of male sex organs.
Alterations in testis weight were inconsistent with some studies reporting Pb-induced decreases in testis
weight and some reporting that testis weight was unaffected by Pb treatment. However, of the studies that
reported on this outcome, only those that dosed prior to weaning reported that Pb treatment reduced testis
weight, suggesting that this outcome may be more sensitive to developmental exposures. Few studies
investigated the effects of Pb on accessory sex organ weight in males, and of the few studies available, no
effects of Pb were reported on weight of the prostate, seminal vesicles, or epididymides. Testicular
histopathology was consistently altered by Pb exposure, often resulting in visible changes to the
seminiferous tubules and surrounding tissue. Toxicological studies also reported Pb-induced changes in
cellular structures in the epididymides.

In summary, the collective body of evidence is sufficient to conclude a causal relationship
exists between Pb exposure and male reproductive function. There is coherent evidence across the
epidemiologic and toxicological studies of detrimental effects of Pb exposure on male reproductive
function (Table 8-1). The strongest evidence of effects of Pb on male reproductive function is seen in the
consistency of the reported effects of Pb on sperm and semen parameters in both toxicological and
epidemiologic studies. However, the recent epidemiologic and toxicological studies suggest that Pb
exposure may also result in alterations in testosterone levels, fertility, and changes in morphology or
histology of male sex organs. Epidemiologic studies consistently report associations between Pb
measured in blood and decreased sperm/semen production and quality, and toxicological studies
consistently report Pb-induced reductions of a variety of semen parameters such as sperm density,
motility, viability, and normal sperm morphology. There are biological plausible pathways through which
Pb exposure may alter sperm/semen production and quality. Specifically, Pb exposure has been shown to
cause oxidative stress, which can damage the supportive somatic cells in the testis (Leydig cells and

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Sertoli cells) as well as damage the sperm cells directly. Supportive somatic cells are responsible for
producing sex steroid hormones and regulating spermatogenesis, and disruption of either of these
functions can impact the quality and quantity of the sperm produced.

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Table 8-1 Summary of evidence contributing to causality determinations for Pb exposure and reproductive
and developmental effects

Rationale for Causality
Determination3

Key Evidence13

Key References'3

Pb Biomarker Levels Associated with
Effects0

Effects on Pregnancy and Birth Outcomes - Likely to be Causal

A few high-quality
epidemiologic studies of Pb
levels and preterm birth
demonstrate associations

Evidence in a single quasi-experimental study and
some high-quality epidemiologic studies
demonstrates associations with preterm birth. There
is uncertainty related to exposure patterns resulting in
likely higher past Pb exposure, especially among
maternal Pb levels.

Jelliffe-Pawlowski et al.
Viqeh et al. (2011)

See Section 8.3

Maternal BLLs: >10 [jg/dL

A few high-quality
epidemiologic studies show
associations with relevant
BLLs, but findings are
overall inconsistent

Inconsistent findings for studies for maternal health
outcomes, prenatal growth, birth defects,
spontaneous abortion and pregnancy loss, and
placental function. There is uncertainty related to
exposure patterns resulting in likely higher past Pb
exposure, especially among maternal Pb levels.
Additional uncertainties regarding biomarker of
exposure (maternal blood, maternal serum, maternal
bone, maternal erythrocytes, cord blood, cord blood
serum, placental tissue) and the critical window of
exposure.

See Section 8.3

Maternal BLLs: 0.32-6.7 pg/dL
Cord blood Pb: 0.37-10.78 pg/dL

Inconsistent toxicological
evidence

Previous studies report reduced BW, but recent
studies report few impacts of Pb on BW, abortion, still
birth, maternal weight gain, birth defects, or placental
weight and histology.

See Section 8.3

Placental weight altered at BLLs as low
as 12.42 pg/dL

Effects on Development - Causal

Delayed Puberty Onset

Consistent associations
with relevant BLLs in high-
quality epidemiologic
studies

Consistent evidence in multiple cross-sectional and
longitudinal epidemiologic studies for females and
males. Most of these studies have large sample sizes

See Section 8.4.2.1
Section 8.4.3.1

and	Female Puberty

BLLs: 0.65-6.57 pg/dL

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Rationale for Causality
Determination3

Key Evidence13

Key References'3

Pb Biomarker Levels Associated with
Effects0



and controlled for potential confounding by covariates
such as age and BMI.



Male Puberty
BLLs: 0.66-6.5

Consistent toxicological
evidence with relevant Pb

Consistent toxicological evidence from multiple
laboratories of delayed male and female puberty

Pine et al, (2006)
lavicoli et al, (2006)

Markers of pubertal onset reduced in
animals with BLLs as low as 12.7 [jg/dL

exposures

onset with Pb exposure via diet, drinking water, or
oral gavage in rodents

Dumitrescu et al, (2008a)





Ronis et al, (1998a)







Ronis et al, (1998c)







Dearth et al, (2002)







Dearth et al, (2004)







Ronis et al, (1996)







lavicoli et al, (2004)



Sokol et al. (1985)

Evidence clearly describes
biological plausibility

Toxicological evidence supports hypothalamic-
pituitary-gonadal axis dysfunction and changes in
IGF-1 contributing to Pb-induced delay in puberty
onset.

Pine et al

Dearth et al.

Pine et al. (2006) reported dam BLLs to
be 39.8 [jg/dL at the time of mating;
Dearth et al. (2002) reported dam BLLs
to be 25.4 [jg/dL at weaning

Postnatal Growth

Available epidemiologic Multiple studies, mostly cross-sectional, for children See Section 8.4.1.1	Maternal BLLs: 0.5-10.1 [jg/dL

evidence is inconsistent of varying ages have reported inconsistent results for	Qorcl b|ooc| p|y o 91-3 1 (jg/dL

the association between BLLs and various measures
of growth. There is uncertainty related to exposure
patterns resulting in likely higher past Pb exposure,
especially among maternal Pb levels. Additional
uncertainties regarding biomarkers of exposure
(maternal blood, maternal serum, maternal bone,
maternal erythrocytes, cord blood, cord blood serum,
placental tissue) and the critical window of exposure.

Available toxicological There are inconsistent findings in the toxicological See Section 8.4.1.2	BLLs ranged from 0.0318-29.16 [jg/dL

evidence is inconsistent literature on Pb exposure and postnatal growth.

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Rationale for Causality
Determination3

Key Evidence13

Key References'3

Pb Biomarker Levels Associated with
Effects0

Effects on Female Reproductive Function - Likely to be Causal

A few high-quality
epidemiologic studies of Pb
levels and hormones
demonstrate associations

Evidence in some high-quality cross-sectional
epidemiologic studies demonstrates associations with
hormone levels but results are mixed based on the
hormone examined. There is uncertainty related to
exposure patterns resulting in likely higher past Pb
exposure.

Krieg and Feng (2011)
Chen et al.

I op pf o I

BLLs: 1.6-4.1 pg/dL

A few high-quality
epidemiologic studies of Pb
levels and menopause
demonstrate associations

Evidence in some high-quality epidemiologic studies
demonstrates associations with menopause. There is
uncertainty related to exposure patterns resulting in
likely higher past Pb exposure.

Mendola et a

Eum et a

(2013)

BLLs: 1.21-3.0 pg/dL

Bone Pb
Tibia: 10 pg/g
Patella: 12 pg/g

Available epidemiologic
studies of Pb levels and
fertility are inconsistent

Epidemiologic studies of this association are limited
by the small sample sizes included in those studies.
In addition, most of the study populations were drawn
from women undergoing IVF and/or attending
infertility clinics. There is uncertainty related to
exposure patterns resulting in likely higher past Pb
exposure.

See Section 8.5.2.1

BLLs: 0.50-2.13 pg/dL

Available toxicological
evidence is inconsistent.

Recent toxicological evidence is scarce and reports
no effects of Pb on litter size and number of litters in
exposed dams. Previous evidence reports
inflammation, decreased ovarian antioxidant capacity,
altered ovarian steroidogenesis.

See Section 8.5

See Section 4.8.4 from U.

EPA (2013)

BLLs ranged from 7.72-12.61 pg/dL in
dams in recent literature

Effects on Male Reproductive Function - Causal

Sperm/Semen Production, Quality, and Function

High-quality and consistent
toxicological evidence with
relevant Pb exposures to
rule out chance, bias, and
confounding with
reasonable confidence.

Decreased sperm counts, decreased sperm
production rate, dose-dependent suppression of
spermatogenesis in rodents with drinking water Pb
exposure.

See Section 4.8.3.1 from U.S.

EPA (2013)

See Section 8.6.1

BLL after adult drinking water exposure
for 30 d: 34 pg/dL

BLL after peripubertal or adult drinking
water exposure for 30 d: 35 and 37 pg/dL.

8-76


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Rationale for Causality
Determination3

Key Evidence13

Key References'3

Pb Biomarker Levels Associated with
Effects0



Ultrastructural and histological damage to non-human
primate testis and seminiferous tubules

Maximum BLLs after daily oral Pb
exposure (gelatin capsule) during infancy,
post infancy, or over a lifetime (up to 10
yr): 32 to 36 pg/dL



Histologic damage to rodent seminiferous tubules
including spermatids and developing sperm.

BLL after adult exposure (oral gavage) for
3 mo: 5.31 pg/dL



Ultrastructural abnormalities to rat spermatogenesis.

BLL after i.p. injection for 16 d: 7.4 pg/dL



Direct effects on rodent sperm DNA after drinking
water Pb exposure.

BLL after adult exposure for 13 wk: 19
and 22 pg/dL



Sperm from Pb exposed rats used for IVF of eggs
harvested from unexposed females yielded lower
rates of fertilization.

BLL after adult exposure for 14-60 d: 33-
46 pg/dL



Semen and sperm quality in rabbits with
subcutaneous Pb treatment; ultrastructural damage
to spermatids with i.p. injection of Pb.

BLL after adult exposure for 15 wk: 16-
24 pg/dL

Available epidemiologic
evidence is inconsistent

The few epidemiologic studies examining this See Section 8.6.1.1

outcome generally have small samples sizes and are

drawn from men attending infertility clinics. There is

uncertainty related to exposure patterns resulting in

likely higher past Pb exposure and biomarker of

exposure (blood, semen, seminal plasma, seminal

fluid).

BLLs: 2.18-3.26 pg/dL

Available toxicological
evidence consistently
reports alterations of sperm
and semen parameters

Consistent reductions of sperm with normal See Section 8.6.1
morphology, sperm density, and sperm viability.

BLLs ranged from 5.09-11.8 pg/dL at
time of outcome assessment

8-77


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Rationale for Causality
Determination3

Key Evidence13

Key References'3

Pb Biomarker Levels Associated with
Effects0

Hormone Levels

A few high-quality
epidemiologic studies of Pb
levels and hormones
demonstrate associations

Evidence in some high-quality cross-sectional
epidemiologic studies demonstrates associations with
testosterone levels and adult males, but inconsistent
associations with other hormones. A longitudinal
study among male adolescents reported null
associations with hormone levels.

See Section 8.6.2.1

Concurrent BLLs: 1.0-4.4 [jg/dL

Available toxicological
evidence is inconsistent

Evidence for testosterone is inconsistent across
studies and few studies are available for other male
sex hormones.

See Section 8.6.2

See Section 4.8.3.2 from U.S.

EPA (2013)

Recent study reported effects at BLLs of
5.09 and 19.1 pg/dLattime of outcome
assessment

Fertility

Lack of large, well-
conducted epidemiologic
studies but overall
inconsistent evidence

The few epidemiologic studies examining this
outcome generally have small samples sizes and are
drawn from men attending infertility clinics. There is
uncertainty related to exposure patterns resulting in
likely higher past Pb exposure and biomarker of
exposure (blood, semen).

See Section 8.6.3.1

BLLs: 1.03-1.27 pg/dL

Limited toxicological	Few toxicological studies investigate male fertility, but See Section 8.6.3	Recent study reported effects at BLLs of

evidence	most report reductions in fertility outcomes such as	gee section 4 8 3 3 from U S 9 4 pg/dL

fertilized oocytes in recent literature and number of	(2013)

offspring per litter in previous studies

BLL = blood lead level; BMI = body mass index; BW = birth weight; d = day(s); IGF-1 = insulin-like growth factor 1; IVF = in vitro fertilization; mo = month(s); Pb = lead; wk = week(s);
yr = year(s).

"Based on aspects considered in judgments of causality and weight of evidence in causal framework in Table I and Table II of the Preamble to the ISAs (U.S. EPA. 2015).
bDescribes the key evidence and references, supporting or contradicting, contributing most heavily to causality determination and, where applicable, to uncertainties or
inconsistencies. References to earlier sections indicate where the full body of evidence is described.

°Describes the Pb biomarker levels at which the evidence is substantiated.

8-78


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8.9

Evidence Inventories - Data Tables to Summarize Study Details

Table 8-2 Epidemiologic studies of exposure to Pb and maternal health outcomes

Reference and

Study Design Study PoPulatlon

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Gestational Diabetes Mellitus

Shapiro et al. (2015) MIREC
n: 1274

Canada
2008-2011
Cohort

Women at least 18 yr of
age during the first
trimester of pregnancy
(6 to <14 wk gestation)
with singleton, live births

Blood

Maternal blood was
measured by ICP-MS

Age at Measurement:
Maternal age during first
trimester of pregnancy

Geometric mean:

Normal glucose: 0.6 [jg/dL
IGT cases: 0.6 [jg/dL
GDM cases: 0.6 [jg/dL

Quartiles (|jg/dL):

Q1
Q2
Q3
Q4

0.2-0.4
0.5-0.6
0.6-0.9
0.9—4.1

Maternal health during
pregnancy: GDM

IGT and GDM were
assessed by chart review
based on the results of a
50-g glucose challenge test
and 75 or 100-g OGTT

Age at outcome:

Maternal age at IGT or
GDM diagnosis during
pregnancy

Logistic regression models OR (95% CI):
were adjusted for maternal GDM vs norma| g|UCOse
age, race, pre-pregnancy
BMI, and education

Q1
Q2
Q3
Q4

Reference
0.8 (0.3, 1.9)
0.6 (0.2, 1.6)
1.1 (0.5, 2.6)

p for trend: 0.87

IGT vs. normal glucose

Q1
Q2
Q3
Q4

Reference
0.8 (0.4, 1.8)
0.6 (0.2, 1.3)
0.9 (0.4, 2.1)

p for trend: 0.62

GDM or IGT vs. normal
glucose

Q1
Q2
Q3
Q4

Reference
0.8 (0.4, 1.5)
0.6 (0.3, 1.1)
1.0 (0.6, 1.8)

p for trend: 0.76

8-79


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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Soomro et al. (2019) Etude des Determinants Blood

Poitiers and Nancy
France

February 2003 to
January 2006

Cohort

pre et post natals du
developpement de la
sante de I'Enfant study
n: 623

Pregnant women
between 24 and 28 wk
of gestation

Maternal blood measured
by EAAS with Zeeman
background correction

Age at Measurement:
Maternal age at 24-28 wk
gestation

Geometric mean0:
1.62 [jg/dL
Median0: 1.7 pg/dL
75th°: 2.2 pg/dL
95thc: 3.8 pg/dL
Maxc: 8.0 pg/dL

Maternal health during
pregnancy: GDM

At 24-28 wk, maternal
blood glucose
concentrations were
measured 1 hr after a 50 g
glucose challenge. The
GDM was diagnosed by
using the OGTT when there
were >2 blood glucose
concentrations greater than
the following cut points:
fasting = 95 mg/dL, at

1	hr = 180 mg/dL, at

2	hr= 155 mg/dL, and at

3	hr = 140 mg/dL

Age at outcome:

Maternal age at 24-28 wk
gestation

Multiple logistic regression
models were adjusted for
maternal smoking,
maternal age, maternal
BMI, maternal education
level, pregnancy-induced
hypertension, and number
of siblings

OR (95% CI):

GDM vs. normal glucose:
1.318 (0.895, 1.94)

IGT vs. normal glucose:
0.853 (0.676, 1.077)
GDM or IGT vs. normal
glucose: 0.86 (0.682, 1.084)

Oquri et al. (2019)
Japan

January 2011 to
March 2014

Cohort

JECS
n: 16,955

Pregnant women from
15 Regional Centers
throughout Japan who
had single pregnancies,
did not have a history of
diabetes, or receive
insulin treatment, and
hypoglycemic agents
during pregnancy; did
not use steroids during
pregnancy

Blood

Maternal blood was
measured by ICP-MS
Age at Measurement:
Maternal age at 22 to 28 wk
of gestation

Geometric mean

non-GDM: 6.05 ng/g

GDM: 6.13 ng/g
Max: 70.9 ng/g

Quartiles (ng/g):

Q1: <5.00
Q2: 5.1-10.0

Maternal health during
pregnancy: GDM

Pregnant women were
diagnosed with GDM if the
results of a 75 g, 2 hr
OGTT exceeded:
fasting = 92 mg/dL
(5.1 mmol/L);

1	hr = 180 mg/dL
(10.0 mmol/L); and

2	hr= 153 mg/dL
(8.5 mmol/L)

Age at outcome:
maternal age at diagnosis
of GDM

Logistic regression models
adjusted for maternal age
at birth, pre-pregnancy
BMI, pregnancy-induced
hypertension, and pack-
years in the nulliparous
models and maternal age
at birth, pre-pregnancy
BMI, history of GDM,
pregnancy-induced
hypertension, and pack-
years in the parous
models; Model 1 was a
multi-pollutant model with
both Cd and Pb; Model 3
was a single pollutant
model of Pb

OR (95% CI):



Model 1:



Nulliparous:



Q1

Reference



Q2

1.22 (0.75,

1.97)

Q3

1.60 (0.72,

3.55)

Q4

2.51 (0.72,

8.72)

Parous:



Q1

Reference



Q2

0.88 (0.65,

1.20)

Q3

0.79 (0.41,

1.41)

Q4

0.31 (0.04,

2.29)

Model 3:

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Q3: 10.1-15.0

Nulliparous:

Q4: >15.1

Q1

Reference



Q2

1.19 (0.74, 1.91)



Q3

1.55 (0.70, 3.42)



Q4

2.42 (0.70, 8.40)



Parous:



Q1

Reference



Q2

0.89 (0.66, 1.20)



Q3

0.75 (0.41, 1.39)



Q4

0.30 (0.04, 2.23)

Wang et al. (2019)

Taiyuan
China

2012-2016

Case-control

n: 776 cases and 776
controls

Women aged 18 yr or
older with GA of 20 wk
or more and without
mental illness were
eligible for the study.
Women who had
stillbirths or birth
defects, who had
multiple births, who did
not donate blood
samples, or who had
gestational weeks less
than 29 wk were
excluded

Blood

Maternal blood was
measured by ICP-MS

Age at Measurement:

Mean maternal age for
GDM: 31.00 yr

Mean maternal age for non-
GDM: 30.97 yr

Median0: 2.7968 [jg/dL
75thc: 3.5981 pg/dL

Tertiles (pg/dL):

Low: <2.254
Middle: 2.254-3.323
High: >3.323

Maternal health during
pregnancy: GDM

GDM diagnosis was based
on a 75 g OGTT during
gestational weeks 24 and
28. Women who met one or
more of the following
criteria were diagnosed
with GDM: (1) fasting blood
glucose was more than
5.1 mmol/L, (2) 1 hr blood
glucose >10.0 mmol/L, or
(3) 2 hr blood glucose
>8.5 mmol/L

Age at outcome:
maternal age at gestational
weeks 24-28

Logistic regression models
adjusted for pre-
pregnancy BMI,
gestational weight gain,
physical activity, parity,
family history of diabetes,
and month of conception;
the multi-pollutant model
was also adjusted for
nickel, As, Cd, antimony,
thallium, Hg, and Pb

OR (95% CI):

Single Pollutant Pb Model:
Low: Reference
Middle: 1.04 (0.81, 1.35)
High: 1.01 (00.78, 1.30)
p for trend: 0.963

Multi-pollutant Model:
Low: Reference
Middle: 1.06 (0.80, 1.41)
High: 110 (0.80, 1.51)
p for trend: 0.622

Zhou et al, (2021b) n: 8169

Blood

Maternal health during

Logistic regression

OR (95% CI)





pregnancy: GDM

analyses: Model 1

Model 1:

China Pregnant women of GA
in the first trimester
(<14 wk) and singleton

Maternal (serum) analyzed
by polarography method

GDM was diagnosed by the
75 g OGTT according to

adjusted for maternal age,
parity, first trimester BMI,
history of spontaneous

T1: Reference
T2: 1.05 (0.90, 1.21)



abortion, history of ectopic



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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

January 2017-
December 2018

Cohort

pregnancy with no
diabetes prior to
pregnancy were
recruited from their first
prenatal visit to the
Southern Medical
University Affiliated
Foshan Women and
Children's Hospital.

Age at measurement:
maternal mean age:
30.14 yr

Median20: 2.53 |jg/dL
75thc: 4.00 pg/dL

Tertiles (pg/dl_):

T1
T2
T3

<1.96

1.961-3.41

>3.411

the International
Association for Diabetes in
Pregnancy Study Group's
criteria.

Age at outcome: maternal
mean age: 30.14 yr

pregnancy, family history
of diabetes, family history
of hypertension; Model 2
adjusted for Model 1 plus
other five (Mn, copper,
calcium, zinc, and
magnesium) metals

T3: 0.89 (0.76, 1.03)

Model 2:

T1
T2
T3

Reference
1.05 (0.90, 1.22)
0.89 (0.76, 1.04)

Zheng et al. (2021)
Boston,

Massachusetts
United States

1999-2002

Cohort

Project Viva
n: 1311

Pregnant women
participating in Project
Viva were included in
this study; women were
those of singleton
gestation, able to
answer questions in
English, and GA <22 wk
at recruitment.

Blood

Maternal blood
(erythrocyte) measured in
the first trimester measured
by ICP-MS

Age at measurement:
maternal age during first
trimester mean (SD): 32.3
(4.6) yr

Median: 17.6 ng/g
75th: 23.6 ng/g

Maternal health during
pregnancy: gestational
glucose

Glucose tolerance test 26-
28 wk gestation, as
measured by non-fasting
50 g oral glucose challenge
test.

Age at outcome: maternal
age at 26-28 wk gestation
mean (SD): 32.3 (4.6) yr

BKMR models adjusted
for maternal age, self-
identified race/ethnicity,
pre-pregnancy BMI, GDM
in prior pregnancy,
smoking, maternal
education, diabetes status
of biological mother, and
gestational week at blood
collection for metals
measurements

Difference in mid-gestational
glucose concentration
(mg/dL) associated with IQR
changes of Pb exposure, with
all other metals fixed at their
medians (95% credible
interval)15: -0.5 (-1.6, -0.6)

Tatsuta et al.

Japan

2011-2014

Cohort

JECS
n: 78,964

Women who delivered a
live infant with singleton
pregnancy. Women
were excluded if there
was a missed blood
sample, missed

Blood

Maternal blood measured
by ICP-MS

Age at measurement:

Maternal age at second or
third trimester; non-GDM

Maternal health during
pregnancy: GDM

GDM diagnosed by OGTT
in second or third trimester
overt GDM diagnosed prior
to OGTT was excluded.

Age at outcome: maternal

Logistic regression
adjusted for pre-
pregnancy BMI, age at
blood collection,
smoking/drinking habits
during pregnancy, history
of GDM, history of
delivering a macrosomia,
regional center, fish intake

OR (95% CI):

Q1
Q2
Q3
Q4
Q5

Reference
1.026 (0.872,
0.968 (0.821,
1.007 (0.854,
0.974 (0.824,

1.206)
1.141)
1.187)
1.151)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

diagnosis of GDM,
missing HbA1c data,
HbA1c >6.5% at <24
gestational weeks, or a
history of type 1 or type
2 diabetes.

mean age 31.0 yr; GDM
mean age 33.3 yr

Median: 5.9 ng/g
95th: 10.6 ng/g

age at second or third
trimester; non-GDM mean
age 31.0 yr; GDM mean
age 33.3 yr

and co-exposure to Cd,
Mn, and Se

Epigenetic Effects During Pregnancy

Sanders et al

Mexico City
Mexico

2007-2011

Cohort

PROGRESS birth cohort
n: 60

This study was
conducted on a sub-
cohort of 60 Mexican
women aged 18-40 yr
participating in the
PROGRESS birth cohort
in Mexico City, and who
consented to a cervical
swab during mid-
pregnancy (16-19 wk
gestation) for miRNA,
thereby participating in
the PROGRESS Cervix
Study.

Blood and bone

Maternal blood was
measured with a dynamic
reaction cell ICP-MS.
Maternal bone was
measured with spot-source
109Cd K-XRF instrument
within 1 mo of delivery

Age at Measurement:
Maternal age at exposure
sampling (mean 27.9 yr
with a range of 18-40)

Mean:

Blood: 2.85 pg/dL
Patellad: 4.16
Tibiad: 1.45
Max:

Blood: 9.38 pg/dl_

Patella3: 20.90
Tibia3: 19.45

Maternal health during
pregnancy: altered miRNA
expression in the cervix

Cervical cells were
collected in a method
similar to a standard Pap
smear protocol, where a
cotton swab was used to
collect cells from the
endocervix. Total RNAwas
extracted using the Exiqon
miRCURY kit. MiRNAs
were quantified by using a
NanoPhotometer P-300.
MiRNA expression was
assessed using the
NanoString nCounter
system.

Age at outcome:

Maternal age at
assessment (mean 27.9 yr
with a range of 18-40)

Multivariable linear	(3 (95% Cl)b, interpreted as %

regression models were expression change
adjusted for maternal age,

education, smoke

Blood, per 10-fold increase in

exposure in the home, and p^.

panty	hsa-miR-297: 84.0 (15.7,

192.8)

hsa-miR-188: 48.5 (7.9,
1.04.2)

Bone Pb, per 1 -unit increase

in Pb:

Patella:

hsa-miR-320e: -4.7 (-7.3,
-1.4)

hsa-miR-22-3p: -4.7 (-8.6,
-0.7)

hsa-miR-93-5p: -6.7 (-12.3,
-0.7)

hsa-miR-769-5p: -5.4 (-9.9,
-0.7)

hsa-miR-297: 2.1 (0.0, 4.2)

hsa-miR-425-5p: -6.7
(-12.3, 0.0)

hsa-miR-361-3p: 2.8 (0.0,
5.7)

Tibia:

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

hsa-miR-575: -4.1 (-6.7,
-1.4)

hsa-miR-4286: -8.6 (-13.5,
-3.4)

hsa-miR-15a-5p: 7.2 (1.4,
14.1)

hsa-miR-142-3p: 5.7 (0.7,
11.7)

hsa-miR-193b-3p: -7.3
(-12.9, -0.7)

hsa-miR-494: -4.1 (-8.0, 0.0)

Sanchez-Guerra et

al. (2019)

Mexico City, Mexico

December 2007-
2011

Cohort

¦July

PROGRESS Study

n: 410 mother-infant
pairs

Participants who were
<20 wk gestation;
maternal age of >18 yr
and without medical
history of heart or kidney
disease) who underwent
clinical examinations at
different hospitals from
Mexican Social Security
System

Blood

Maternal blood (collected at
second and third trimester
and delivery) and umbilical
cord blood were measured
by ICP-QQQ

Age at measurement:
maternal age at
measurement (mean age
27.22 yr)

Mean

second trimester:

3.79 [jg/dL
3.90 [jg/dL
4.16 [jg/dL
3.50 [jg/dL

75th

second trimester:

third trimester:
at delivery:
cord blood:

4.51 [jg/dL
4.73 [jg/dL
5.28 [jg/dL
4.45 [jg/dL

third trimester:
at delivery:
cord blood:

Maternal health during
pregnancy: altered cord
blood mtDNA content

Venous cord blood
measured the relative
mtDNA content through
mitochondrial-to-nuclear
DNA ratio in cord blood

Age at outcome:
Maternal age at delivery
(Mean age 27.22 yr)

Multivariate linear
regression models were
adjusted for sex, mother's
age, mother's BMI, SES,
smoke exposure, PM2.5
levels, GA, platelets and
leucocytes in cord blood,
C-section, PROM,
preeclampsia, and date of
visit

(3 (95% CI)

Maternal blood
Second trimester: 0.017
(0.002, 0.031)

Third trimester: 0.015 (0.00,
0.03)

At delivery: 0.013 (-0.001,
0.027)

Cord blood

At delivery: 0.016(0.001,
0.03)

8-84


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Study"Desfg".? Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Other Outcomes Related to Maternal Health During Pregnancy

Kahn et al. (2014)

Mitrovoca and

Pristina,

Kosovo

May 1985 and
December 1986

Cohort

Yugoslavia Prospective
Study of Environmental
Lead Exposure
n: 291

Women in their second
trimester of pregnancy
were invited to
participate in a study of
pregnancy outcomes at
their first prenatal visit to
government clinics
located at the centers of
two towns in Kosovo.
Women with singleton
births, between 18 and
44 wk of gestation, had
no major central nervous
system defects, no
chromosomal
abnormalities, and
residing <10 km from
clinic

Blood

Maternal blood (serum)
collected at mid-pregnancy
(no method reported)

Age at Measurement:
Pristina Mean: 26.6 yr;
Mitrovica Mean: 26.7 yr

Mean:

Pristina: 5.57 [jg/dL;
Mitrovica: 20 [jg/dL
Max:

Pristina: 18.60 pg/dL;
Mitrovica: 41.30 [jg/dL

Maternal health during
pregnancy: thyroid function
during pregnancy

Maternal thyroid function
during pregnancy was
assessed using 1T4, TSH,
and TPOAb. 1T4 and
TPOAb were measured by
a radioimmunoassay
procedure, and TSH was
measured using an
immunoradiometric assay
procedure.

Age at outcome:

Pristina Mean: 26.6 yr;
Mitrovica Mean: 26.7 yr

Multiple linear regression
analysis: 1T4 models
adjusted for height,
ethnicity, BMI, fetal GA,
maternal education, adults
per room; TSH models:
hemoglobin, ethnicity,
BMI, fetal GA, maternal
age; TPOAb models
(continuous and
dichotomous): ethnicity,
fetal GA, maternal age,
adults per room

(3 (95% Cl)b

1T4: -0.074 (-0.10, -0.046)
TSH: 0.026 (-0.065, 0.12)
TPOAb: 0.31 (0.17, 0.46)

OR (95% Cl)b

TPOAb: 2.41 (1.53, 3.82),
comparing >10 lU/mLvs.
<10 lll/mL

Wells et al. (2011)

Baltimore, MD
United States

November 2004 and
March 2005

Cohort

Baltimore Tracking
Health Related to
Environmental
Exposures Study
n: 285

Singleton births with
cord blood available,
with complete covariates
data

Blood

UCB was measured by
ICP-MS

Age at Measurement:
Maternal age at delivery
(range: 14-43, mean: 26)

Geometric mean:
0.66 [jg/dL
75th: 0.96 pg/dL
Max: 6.47 pg/dL

Maternal health during
pregnancy: blood pressure
in late pregnancy

Hospital personnel
measured maternal blood
pressure at admission for
labor and delivery and
continuously during
hospitalization. Three pairs
of blood pressure
measurements from each
mother were recorded: SBP
and DBP at admission, the

Multivariable linear
regression models were
adjusted for maternal age,
maternal race,
neighborhood median
household income, prima
parity, smoking during
pregnancy, pre-pregnancy
BMI, and anemia

(3 (95% CI), as change in
blood pressure (mmHg)

Admission SBP:

Q1: Referent

Q2 2.89 (-2.16, 7.94)

Q3: 1.05 (-4.04, 6.14)

Q4: 6.87 (1.51, 12.21)

p for trend: 0.033

Admission DBP:
Q1: Referent

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

maximum SBP and

Q2

0.00 (-3.95, 3.96)

corresponding DBP, and

Q3

0.81 (-3.17, 4.80)

the minimum SBP and

Q4

corresponding DBP.

4.40 (0.21, 8.59)



p for trend: 0.036

Age at outcome:





maternal age at delivery

Maximum SBP:

(range: 14-43, mean: 26)

Q1

Referent



Q2

2.47 (-3.08, 8.02)



Q3

-1.76 (-7.36, 3.85)



Q4

7.72 (1.83, 13.60)



p for trend: 0.055



Maximum DBP:



Q1

Referent



Q2

3.93 (-2.86, 10.72)



Q3

-0.42 (-7.27, 6.43)



Q4

8.33 (1.14, 15.53)



p for trend: 0.086

Li eta!. (2017b)

Shanghai
China

2010

Cohort

N: 1,485

Pregnant women during
late pregnancy (28-36
gestational weeks)

Blood

Maternal blood was
measured by background
corrected GFAAS collected
gestational week 28-36

Age at measurement:
42 yrold

Geometric mean:
3.99 [jg/dL

Max: 14.84 pg/dL

13-

Maternal health during
pregnancy: maternal stress

Maternal life event stress
and emotional stress were
assessed using the LESPW
and SCL-90-R,
respectively.

Generalized additive
models were adjusted for
maternal age, ethnicity,
maternal education, family
monthly income, years
living in Shanghai

Age at outcome:
old

13-42 yr

(3 (95% Cl)b

Log-blood Pb
GSI: 0.01 (-0.05, 0.07)
Depression: 0.03 (-0.05,
0.10)

Anxiety: 0.01 (-0.06, 0.08)

Log-blood Pb <0.41 [jg/dL
GSI: 0.22 (0.05, 0.40)
Depression: 0.34 (0.12, 0.56)
Anxiety: 0.01 (-0.06, 0.08)

Log-blood Pb >0.41 [jg/dL

8-86


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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

GSI: -0.07 (-0.16, 0.01)

Depression: -0.09 (-0.19,
0.02)

Anxiety: -0.08 (-0.18, 0.02)

Osorio-Yanez et al.

(2021)

Mexico

2007-2011

Cohort

PROGRESS
n: 668

Women enrolled during
second trimester of
pregnancy, were >18 yr
of age, lived in Mexico
City for the following 3 yr

Blood and bone

Maternal blood was
measured by ICP-QQQ;
bone Pb measured by
K-XRF and obtained two
estimated for patella and
tibia (one for each leg),
which were measured 26-
55 d postpartum

Age at measurement:
Median (SD): 27(5.5)yr

Median

Blood - 2nd Trimester:
2.80 [jg/dL

Blood - 3rd Trimester:
2.99 [jg/dL
Bone, tibia: 2.84 |jg/g
Bone, patella: 3.49 |jg/g

Max:

Blood:

2nd Trimester: 20.70 [jg/dL
3rd Trimester: 28.25 [jg/dL
Tibia: 30.1 |jg/g
Patella: 43.2 |jg/g

Maternal health during
pregnancy: bone
remodeling

Bone speed of sound
measured at the second
and third trimesters of
pregnancy at the distal
radius and medium
phalange using QUS.

Age at outcome: Median
(SD): 27 (5.5) yr

Linear models adjusted for
maternal age, SES, parity,
BMI, and GA at the time of
Z-score measurement;
linear mixed model
adjusted for maternal age,
SES, parity, BMI, and GA
at the time of QUS
measurement; models
with blood were mutually
adjusted for other (Cd and
As) metals

(3 (95% Cl)b

Bone (radius) QUS Z-score at
2nd Trimester
Blood (|jg/dL): -0.06 (-0.18,
0.07)

Tibia (|jg/g bone mineral):
0.002 (-0.07, 0.07)

Patella (pg/g bone mineral):
-0.08 (-0.15, -0.01)

Bone (radius) QUS Z-score at
3rd Trimester

Blood (|jg/dL): -0.03 (-0.16,
0.10)

Tibia: 0.017 (-0.05, 0.09)
Patella: -0.03 (-0.10, 0.05)

Bone (radius) QUS Z-score
during pregnancy
Blood (|jg/dL): -0.04 (-0.13,
0.04)

Tibia (|jg/g bone mineral):
0.006 (-0.04, 0.06)

Patella (pg/g bone mineral):
-0.06 (-0.10, -0.01)

Kim et a I . (2022)

PROTECT
n: 617

Blood

Maternal health during
pregnancy: MMP

Linear mixed effects
models adjusted for
maternal age, education,

B (95% CI)b as percent
change in MMP per IQR
increase in blood Pb

8-87


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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Puerto Rico and
United States

2010

Cohort

Pregnant women in the
first trimester or early
second trimester of
pregnancy that resided
in the Northern Karst
aquifer region, known
for a large number of
Superfund and other
hazardous waste sites.

Maternal blood, collected at
up to two study visits
(median 18- and 26-wk
gestation), was measured
by ICP-MS

Age at measurement:

Mean (SD) age at
enrollment: 26.9 (5.5) yr

Median:

Enrollment: 0.32 ng/mL
Follow up:0.32 ng/mL
75th:

Enrollment: 0.42 ng/mL
Follow up: 0.43 ng/mL
Max:

Enrollment: 2.18 ng/mL
Follow-up: 1.51 ng/mL

Expression levels of MMP1,
MMP2, and MMP9
measured using
customized Luminex assay
from Invitrogen

Age at outcome: Mean
(SD) age at enrollment:
26.9 (5.5) yr

exposure to second-hand
tobacco smoke, and pre-
pregnancy BMI

MMP1
MMP2
MMP9

23.6 (12.9, 35.2)
5.89 (2.23, 9.67)
-3.31 (-8.12, 1.75)

Females:

MMP1
MMP2
MMP9

Males:

MMP1
MMP2
MMP9

16.3 (5.74, 28.0)
5.48 (1.50, 9.62)
-1.89 (-7.35, 3.90)

10.5 (1.15, 20.6)
2.24 (-1.25, 5.86)
-5.14 (-9.85, -0.17)

Gajewska et al.

(2021)

Poland

2018-2020

Case-control

n: 146 (66 with
preeclampsia)

Healthy pregnant
women and healthy non-
pregnant women visiting
the Independent Public
Clinical Hospital No 4 in
Lublin for a stay in the
hospital or routine
testing.

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
Mean: 29.16 yr
Median: 28 yr
Range: 18-47 yr

All Participants:

Mean (SD): 2.63
(1.34) [jg/dL

Median: 2.6 [jg/dL
Preeclampsia Participants:

Maternal health during
pregnancy: preeclampsia

Diagnosis of preeclampsia
was based on the definition
from the American College
of Obstetrics and
Gynecologists.

Age at outcome:

Mean: 29.16 yr
Median: 28 yr
Range: 18-47 yr

Logistic regression
adjusted by the pregnant
woman's age, place of
resident (urban/rural), GA,
multiplicity of pregnancy,
and number of previous
pregnancies

OR (95% CI)b: 2.65 (1.2,
5.86)


-------
Outcome

Confounders

Effect Estimates and 95%
Clsa

Reference and
Study Design

Study Population

Exposure Assessment

Mean (SD) 3.36 (1.23)
Median: 3.49 [jg/dL

Max:

All Participants: 6.1 [jg/dL
Preeclampsia Participants:
6.1 [jg/dL

Wu etal. (2021)
China,

Foshan, Guangdong
Province

August 2019-
November 2019
(participants followed
from 8-12 wk of
pregnancy to birth)

Cohort

n: 2174

Pregnant women that
were registered,
checkup, and delivering
in the Foshan
Chancheng Central
Hospital were included
in the study.

Blood

Maternal blood, collected
between 12 and 27 (±6) wk
of pregnancy and before
date of diagnosed
preeclampsia, was
measured by AAS

Age at measurement:

Mean age at delivery (SD):
29.04 (4.25) yr

Median: 3.60 [jg/dL

Quartiles (|jg/dL):

Q1
Q2
Q3
Q4

2.00-2.90
3.00-3.60
3.70-4.40
4.50-7.90

Maternal health during
pregnancy: preeclampsia

Preeclampsia was based
on electronic medical
records. Preeclampsia was
defined as newly diagnosed
hypertension and
proteinuria occurring after
20 wk of gestation.
Hypertension was defined
as systolic >140 mmHg or
DBP >90 mmHg, 2
occasions, 4 hr apart in a
previously normotensive
woman. Proteinuria was
defined as >300 mg/24-hr
urine collections, or
protein/creatinine >0.3, or
dipstick reading >1

Age at outcome: maternal
age after 28 wk gestation

Logistic regression models OR (95% CI)
were adjusted for age at
delivery, pre-pregnancy
BMI, parity, method of
conception (natural
conception, ART
conception), and
education level; logistic

Dose-effect analysis of the
relationship between BLLs
and the risk of preeclampsia

Linear regression modelb:
1.43 (1.17, 1.74)

BLLs <4.2 |jg/dLb: 0.79 (0.50,
1.24)

BLLs >4.2 |jg/dLb: 2.05 (1.50-
2.81)

Preeclampsia:

Continuous modelb: 1.43
(1.17, 1.74)

Q1: Reference

Q2
Q3
Q4

1.48 (0.64, 3.39)
0.85 (0.33, 2.20)
2.38 (1.13, 5.03)

p for trend: 0.02

Mild Preeclampsia:

Continuous modelb: 1.62
(1.27, 2.06)
Q1: Reference
Q2: 2.63 (0.81, 8.63)
Q3: 1.33 (0.35, 5.06)
Q4: 4.26 (1.41, 12.89)

8-89


-------
Outcome

Confounders

Effect Estimates and 95%
Clsa

p for trend: 0.01

Reference and
Study Design

Study Population

Exposure Assessment

Severe Preeclampsia:

Continuous modelb: 1.10
(0.72, 1.68)

Q1: Reference

Q2: 0.69 (0.19, 2.49)

Q3: 0.51 (0.13, 2.05)

Q4: 1.12 (0.38, 3.27)

p for trend: 0.78

Braun et al. (2014)

Mexico City,
Mexico

July 2007 and
February 2011

Cohort

n: 1054

Participants for this
study were enrolled from
an ongoing prospective
birth cohort in Mexico
City. Pregnant women
receiving health
insurance and prenatal
care through the
Mexican Social Security
System were invited to
participate in the study.
To be eligible for
participation in the
study, women had to be
<20 wk gestation, >18 yr
old, free of heart or
kidney disease, have
access to a telephone,
plan to reside in Mexico
City for the next 3 yr, not
use steroids (including
glucocorticoids) or anti-
epilepsy drugs, and not
consume alcohol on a
daily basis.

Blood and bone

Maternal blood was
measured by GFAAS
during the second trimester.
Maternal bone was
measured by K-XRF
instrument ~1 mo
postpartum

Age at measurement:
>18 yr old

Mean:

blood: 3.7 [jg/dL
tibia: 2.7 |jg/g
patella: 4.6 |jg/g

Blood Pb Quintiles
Q1
Q2
Q3
Q4
Q5

0—<1.8 |jg/dL
1.8-<2.4 Mg/dL
2.4-<3.4 [jg/dL
3.4-<5.1 pg/dL
>5.1 [jg/dL

Maternal health during
pregnancy: hypothalamic-
pituitary-adrenal axis
function measured from
salivary Cortisol
concentrations

Between 14 and 35 wk of
gestation (mean [SD]: 19.7
[2.4] wk), pregnant women
provided five saliva
samples each day over 2
consecutive days during
the week or weekend.
Women were instructed to
provide samples using the
passive drool technique
upon awakening, 45 min
after waking, 4 hr after
waking, 10 hr after waking,
and at bedtime. Saliva
samples were assayed in
the same batch in duplicate
for Cortisol using a
chemiluminescence assay
with sensitivity of
-0.16 ng/ml.

Linear mixed models with
random intercepts for day
and participant were
adjusted for maternal age,
marital status, years of
education, parity, and
smoking status (never,
former, and current), BMI,
and stress or depressive
symptoms

(3 (95% CI), as % difference in
Cortisol area under the curve
nmol-hr

Blood Pb Quintiles

Q1

Reference

Q2

8 (-1, 18)

Q3

9 (0, 19)

Q4

8 (-1, 18)

Q5

CM
CO
CM

Tibia Pb Quintiles:

Q1

Reference

Q2

-5 (-14, 5)

Q3

2 (-8, 13)

Q4

0 (-10, 10)

Q5

00
CD

Patella Pb Quintiles

Q1

Reference

Q2

1 (-8, 12)

Q3

-6 (-14, 4)

Q4

"3"

CM

CM

8-90


-------
Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Tibia Pb Quintiles

Q1
Q2
Q3
Q4
Q5

<2 pg/g

2-<4.3 |jg/g
4.3-<6.7 |jg/g
6.7—<11.1 |jg/g
>11.1 |jg/g

Age at outcome:
maternal age at the time of
outcome measurement

Q5: 4 (-6, 16)

Patella Pb Quintiles
Q1: <2 |jg/g
Q2: 2-<4.5 |jg/g
Q3: 4.5-<7.8 |jg/g
Q4: 7.8-<12.7 pg/g
Q5: >12.7-43.2 |jg/g

Ishitsuka et al.

Japan

January 2011 -
March 204

Cohort

JECS
n: 17,267

Pregnant women from
15 Regional Centers
throughout Japan who
had single pregnancies,
did not have a history of
diabetes, or receive
insulin treatment, and
hypoglycemic agents
during pregnancy; did
not use steroids during
pregnancy

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
maternal age at 27 wk of
gestation (mean age:
31 ± 5 yr)

Geometric mean:
0.58 [jg/dL

Quintiles (|jg/dL):

Q1
Q2
Q3
Q4
Q5

0.143-0.433

0.444-0.523

0.524-0.616

0.617-0.7533

0.754-6.752

Maternal health during
pregnancy: maternal
depression

Psychological symptoms
during middle or late
pregnancy were assessed
using the K6.

Age at outcome:
maternal age at 27 wk of
gestation (mean age:
31 ± 5 yr)

Multivariable logistic
regression models
adjusted for age, parity,
marital status, education,
employment status,
household income, and
smoking and alcohol
status

OR (95% CI)

Pb per one-unit increase
K6 >13: 1.00 (0.76, 1.32)
K6 >5: 0.98 (0.88, 1.09)

K6 >13:

Q1
Q2
Q3
Q4
Q5

Reference
0.94 (0.69, 1.27)
0.97 (0.71, 1.31)
0.92 (0.68, 1.25)
0.87 (0.64, 1.19)

K6 >5:

Q1

Reference

Q2

1.03 (0.92, 1.16)

Q3

1.07 (0.95, 1.19)

Q4

0.98 (0.87, 1.10)

Q5

1.01 (0.90, 1.13)

8-91


-------
Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Christensen et al.

Ukraine and
Greenland

2002-2004
Cross-sectional

Climate Change,
Environmental
Contaminants, and
Reproductive Health
n: 117

Women at least 18 yr
old and born in the
country of the study.

Blood

Maternal blood was
measured by ICP-MS

Age at measurement: >18

Mean0: 1.74 [jg/dL
Median0: 1.457 [jg/dL

Tertiles°(|jg/dL):

T1
T2
T3

0.544-1.013
1.013-1.902
1.902-14.088

Maternal health during
pregnancy: AMH

Concentrations of AMH
were assessed by the
Immunotech enzyme
immunoassay
AMH/Mullerian-inhibiting
substance assay from
serum.

Age at outcome: >18

General linear models
were adjusted for GA,
maternal age, research
site, parity, fish intake,
BMI, ever smoker and
pelvic diseases and
infections

(3 (95% Cl)b per one-unit In-
Pb increase: -0.0423
(-0.4989, 0.4144)

Gustin et al. (2021)
Sweden,

Norrbotten county

Enrollment: 2015-
2018, follow-up
through 29
gestational weeks

Cohort

NICE
n: 544

Pregnant women visiting
their local maternity
clinics who were
residents of southern or
eastern Norrbotten
count and planned to
give birth at Sunderby
Hospital. Only first
pregnancies and
singleton births
included. Those with
thyroid dysfunction were
excluded.

Blood

Maternal blood
(erythrocyte) was measured
by ICP-MS

Age at measurement:
Median: 30 yr

Median: 11 |jg/kg
95th: 27 pg/kg

Maternal health during
pregnancy: hormone Levels
(1T4, tT4, 1T3, tT3, TSH,
1T4:tT4, 1T3:tT3, 1T3:1T4)

Plasma samples from
gestational week 29
analyzed via

electrochemiluminescence
immunoassays

Multivariate linear
regression models
adjusted for parity,
maternal education,
maternal pre-pregnancy
smoking

(3 (95% Cl)b

1T4 (pmol/L): 0.014 (-0.21,
0.18)

tT4 (nmol/L): 0.90 (-1.5, 3.3)
1T3 (pmol/L): 0.036 (-0.018,
0.090)

tT3 (nmol/L): 0.038 (-0.015,
0.091)

TSH (mlU/L): -0.023 (-0.13,
0.087)

1T4:tT4: -0.001 (-0.002,
0.001)

1T3:tT3: -0.009 (-0.031,
0.014)

1T3:fT4: 0.004 (-0.003, 0.011)

Corrales Vargas et

al. (2022)

n: 344

Blood

Maternal blood measured
by ICP-MS

Maternal health during
pregnancy: thyroid function

TSH, 1T4, and 1T3

Linear regression models
adjusted for age, GA,
cotinine detection, pre-
pregnancy BMI, and

(3 (95% CI) for % change in
1st measurement of
outcomes per 10% increase
in blood Pb (pg/L) at
enrollment

8-92


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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Matina County,
Limon
Coast Rica

2010-2011

Cohort

Age at measurement:
Maternal age at collection
(recruited <33 wk gestation
with 2nd blood sample
10 wk later)

Median: 0.666 [jg/dL
75th: 0.908 pg/dL
90th: 1.211 pg/dL
Max: 3.43 pg/dL

measured in serum using
electrochemiluminescence

severe vomiting during
pregnancy

TSH (mlU/L): -2.3 (-16.15,
11.55)

fT4 (pmol/L): 0.99 (-0.11,
2.09)

1T3 (pmol/L): -0.21 (-0.52,
0.10)

(3 (95% CI) for % change in
2nd measurement of
outcomes per 10% increase
in blood Pb (pg/L) at
enrollment, excluding outliers:
TSH (mlU/L): -0.08 (-0.22,
0.07)

1T4 (pmol/L): 1.96 (0.66, 3.25)
1T3 (pmol/L): 0.24 (-0.13,
0.61)

AAS = atomic absorption spectrometry; AMH = anti-Mullerian hormone; ART = assisted reproductive technology; BKMR = Bayesian kernel machine regression; BMI = body mass index;

Cd = cadmium; CI = confidence interval; d = day(s); DBP = diastolic blood pressure; EAAS = electrothermal atomic absorption spectrometry; fT3 = free triiodothyronine; fT3:fT4 = ratio of

free triiodothyronine to free thyroxine; fT3:tT3 = ratio of free triiodothyronine to total triiodothyronine; fT4 = free thyroxine; fT4:tT4 = ratio of free thyroxine to total thyroxine;

GDM = gestational diabetes mellitus; GFAAS = graphite furnace atomic absorption spectrometry; GSI = Global Severity Index; hr = hour(s); ICP-MS = inductively coupled plasma mass

spectrometry; ICP-QQQ = inductively coupled plasma triple quad; IGT = impaired glucose tolerance; IQR = interquartile range; JECS = Japan Environment and Children's Study;

K6 = Kessler Psychological Distress Scale; K-XRF = K-shell X-ray fluorescence; LESPW = Life Event Scale for Pregnant Women; MIREC = Maternal-Infant Research on Environmental

Chemicals; miRNA = micro RNA; min = minute(s); MMP = matrix metalloproteinases; mo = month(s); mtDNA = mitochondrial DNA; NICE = Nutritional impact on Immunological maturation

during Childhood in relation to the Environment; OGTT = oral glucose tolerance test; OR = odds ratio; PM25 = fine particulate matter; PROGRESS = Programming Research in Obesity,

Growth, Environment and Social Stressors; PROM = premature rupture of membranes; PROTECT = Puerto Rico Test site for Exploring Contamination Threats; Q = quartile;

QUS = quantitative ultrasound; SBP = systolic blood pressure; SCL-90-R = Symptom-Checklist-90-Revised; SD = standard deviation; Se = selenium; SES = socioeconomic status;

TPOAb = thyroid peroxidase antibodies; TSH = thyroid-stimulating hormone; tT3 = total triiodothyronine; tT4 = total thyroxine; UCB = umbilical cord blood; wk = week(s); yr = year(s).

aEffect estimates are standardized to a 1 pg/dL increase in blood Pb or a 10 pg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect estimates are

standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated interval. Categorical

effect estimates are not standardized.

bEffect estimates unable to be standardized.

°Pb measurements were converted from pg/L to pg/dL.

dNo units provided.

8-93


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Table 8-3

Animal toxicological studies of Pb exposure and pregnancy and birth outcomes

Study

Species (Stock/Strain), n,
Sex

Timing of
Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported (pg/dL)

Endpoints
Examined

Saleh eta!. (2018)

Rat (Sprague-Dawley)

Control (vehicle), F, n = 8
dams

160 ppm Pb, F, n = 8 dams
320 ppm Pb, F, n = 8 dams

GD 1 to 20	Dams were dosed via oral

gavage. Authors report a
significant decrease in brain
weight occurred, indicating
potential overt toxicity.

Dams (GD 20):

5.1 [jg/dL for control
27.7 pg/dL for 160 ppm Pb
41.5 pg/dL for 320 ppm Pb

Abortion, Placental
Weight

Saleh etal. (2019)

Rat (Sprague-Dawley)

Control (vehicle), F, n = 8
dams

160 ppm Pb, F, n = 8 dams
320 ppm Pb, F, n = 8 dams

GD 1 to 20	Dams were dosed via oral

gavage. Authors report a
significant decrease in brain
weight occurred, indicating
potential overt toxicity.

Dams (GD 20):

5.26 [jg/dL for control
23.9 pg/dL for 160 ppm Pb
42.9 pg/dL for 320 ppm Pb

Placental Weight

Cory-Slechta et al.

(2013)

Mouse (C57BL/6)

Control (untreated), M/F,
n = 16-29 (8-17/8-12) pups

100 ppm Pb, M/F, n = 16-29
(8-17/8-12) pups

GD -61 to	Dams were dosed via

PND 365	drinking water starting 2 mo

prior to mating. Offspring
were continued on the same
exposure as their dams until
the end of the experiment at
12 mo of age. Sample sizes
are only available for "Final"
group sizes for males and
females in Table 1.

Dams at weaning (PND 24):
0.22 [jg/dL for control
12.12 pg/dL for 100 ppm Pb

BW, Sex Ratio

Schneider et al.

Mouse (C57BL/6)

GD -61 to

Dams were dosed via

Dams at weaning (assumed BW

(2016)

Control (untreated), F,

PND 21

drinking water starting 2 mo

PND 21): 0.22 pg/dL for control



n = NR



prior to mating through

12.61 pg/dL for 100 ppm Pb







lactation (weaning assumed



100 ppm Pb, F, n = NR



to be PND 21).

Pups (PND 5-6): 0.37 pg/dL for
control

10.2 pg/dL for 100 ppm Pb





Dams were also treated to a
non-stress or prenatal stress

8-94


-------
Study

Species (Stock/Strain), n,
Sex

Timing of
Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported ((jg/dL)

Endpoints
Examined







condition. Only data from











dams in the non-stress











condition were used.





Wang et al. (2014)

GD 1-20

Rat (Wistar)	GD 1-10, or

Control (untreated), F, n = 17 ®D	or

dams

0.25% Pb GD 1-10, F,
n = 16 dams

0.25% Pb GD 11-20, F,
n = 15 dams

Dams were dosed via	Dams (assumed GD 20):

drinking water during different	0.828 pg/dL for control

windows of pregnancy.	„„ „„	r,, ^ .

Assumed termination of study	26 29 Mg/dL for 0.25% Pb GD 1-

on GD 20.

Placenta
Histopathology,
Placental Weight

12.4 [jg/dL for 0.25% Pb GD 11-
20

36.02 [jg/dL for 0.25% Pb GD 1-
20

0.25% Pb GD 1-20, F,
n = 15 dams

Weston et al.

Rat (Long-Evans)

Dams

Control (untreated), F, n = 20
50 ppm Pb, F, n = 19
Pups

Control (untreated), M/F,
n = 12.4 (7/5.4 average
number of male and female
pups per litter in control)

50 ppm Pb, M/F, n = 7.4
(6.3/1.1 average number of
male and female pups per
litter in Pb non-stress group)

GD -76 to	Dams were dosed via

PND 21	drinking water starting 2-

3 mo prior to breeding.
Exposure ended at weaning
(PND 21).

Dams (PND 21):

0.500 [jg/dL for control
7.72 |jg/dL for 50 ppm Pb

Pups (PND 5-6):

0.603 [jg/dL for control males
0.690 [jg/dL for control females
15.7 [jg/dL for 50 ppm Pb males
14.6 [jg/dL for 50 ppm Pb females

BW, Sex Ratio

jarkur and Bairy Rat (Wistar)

GD -30 to GD -
GD Oto GD 21;
PND 1 to

Dams were dosed via
drinking water for varying
amounts of time:

Pregestation Only (1 mo prior

Pups (PND 22):
0.19 [jg/dL for control

Stillborn Pups, BW

8-95


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Study

Species (Stock/Strain), n,
Sex

Timing of
Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported (pg/dL)

Endpoints
Examined

Control (untreated), F, n = 6
dams

0.2% Pb Pregestation Only,
n = 6 dams

0.2% Pb Gestation Only,
n = 6 dams

0.2% Pb Lactation Only,
n = 6 dams

0.2% Pb Gestation and
Lactation, F, n = 6 dams

PND21; GD 0 to
PND21

to conception), Gestation
Only (21 d), Lactation Only
(21 d), and Gestation and
Lactation (42 d).

3.03 [jg/dL for 0.2% Pb in
Pregestation Only group

5.51 [jg/dL for 0.2% Pb in
Gestation Only group

26.86 jjg/dL for 0.2% Pb in
Lactation Only group

31.59 pg/dL for 0.2% Pb in
Gestation and Lactation group

jarkur and Bairy

Rat (Wistar)

Control (untreated), F, n = 6
dams

0.2% Pb Pregestation Only,
F, n = 6 dams

0.2% Pb Gestation Only,
F, n = 6 dams

0.2% Pb Lactation Only, F,
n = 6 dams

GD-30toGD-1; Dams were dosed via

GD 0 to GD 21;
PND 0 to
PND21; GD 0 to
PND 21

drinking water for varying
amounts of time:

Pregestation Only (1 mo prior
to conception), Gestation
Only (21 d), Lactation Only
(21 d), and Gestation and
Lactation (42 d).

Pups (PND 22):

0.18 [jg/dL for control

3.02 [jg/dL for 0.2% Pb in
Pregestation Only group

5.30 [jg/dL for 0.2% Pb Gestation
Only group

26.7 [jg/dL for 0.2% Pb in
Lactation Only group

32.0 [jg/dL for 0.2% Pb in
Gestation and Lactation group

Stillborn Pups, BW

0.2% Pb Gestation and
Lactation, F, n = 6 dams

Tartaqlione et al

Rat (Wistar)

Control, M/F, n = NR

50 mg/L Pb, M/F, n = NR

GD -28 to	Dams were dosed via

PND 23	drinking water starting 4 wk

prior to mating until weaning
(PND 23).

Pups (PND 23):

0.700 [jg/dL for 0 mg/L Pb

25.5 [jg/dL for 50 mg/L Pb

BW, Sex Ratio

8-96


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Study

Species (Stock/Strain), n,
Sex

Timing of
Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported (pg/dL)

Endpoints
Examined

Zhaoetal. (2021)

Rat (Sprague-Dawley)

Control (untreated), F, n = 6
dams

109 ppm Pb, F, n = 6 dams

GD-14to	Dams were dosed via

PND 10	drinking water starting 2 wk

prior to mating and continued
until PND 10.

Pups:

PND 0

0.87 [jg/dL for control
48.2 pg/dL for 109 ppm Pb

PND 10

0.87 [jg/dL for control
11.5 pg/dL for 109 ppm Pb

PND 21

0.87 pg/dL for control
2.81 pg/dL for 109 ppm Pb

PND 30

0.87 pg/dL for control
1.20 pg/dL for 109 ppm Pb

BW

Barkur et al, (2011)

Rat (Wistar) GD 0 to PND 21

Control (untreated), F, n = 6
dams

0.2% Pb GD Oto PND 21,

F, n = 6 dams

Dams were dosed via
drinking water throughout
gestation until weaning
(PND 21). Only male pups
were examined.

Pups:

PND 22

0.266 pg/dL for control
31.2 pg/dL for 0.2% Pb

PND 120

0.234 pg/dL for control
0.468 pg/dL for 0.2% Pb

BW

Betharia and Maher
(2012)

Rat (Sprague-Dawley) GD 0 to PND 20

Control (untreated), M/F,
n = 36-48 (18-24/18-24)
pups

10 pg/mL Pb, M/F, n = 36-
48 (18-24/18-24) pups

Dams were dosed via
drinking water throughout
pregnancy until weaning
(PND 20).

Pups:

PND 2

0.188 pg/dL for control
9.03 pg/dL for 10 pg/mL Pb

PND 25:

Stillborn Pups, Sex
Ratio

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Study

Species (Stock/Strain), n,
Sex

Timing of
Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported (pg/dL)

Endpoints
Examined

0.088 [jg/dL for control
0.976 [jg/dL for 10 |jg/mL Pb

PND60:

0.0244 [jg/dL for control
0.0318 [jg/dL for 10 |jg/mL Pb

Graham et al. (2011)

Rat (Sprague-Dawley)

Control (vehicle), M/F,
n = 14-16 (7-8/7-8)

1 mg/kg Pb, M/F, n = 14-16
(7-8/7-8)

10 mg/kg Pb, M/F, n = 14-16
(7-8/78)

PND 4 to 28 Offspring were dosed via oral PND 29:

gavage every other day from 0.267 pg/dL for 0 mg/kg

3.27 pg/dL for 1 mg/kg
12.5 pg/dL for 10 mg/kg

PND 4 until PND 28.

Offspring Mortality

Baranowska-Bosiacka Rat (Wistar)

et al. (2013)

Control (untreated), F, n = 3
dams

0.1% Pb, F, n = 3 dams

Control, M/F, n = 36 (17/19)
pups

0.1% Pb, M/F, n = 36 (18/18)
pups

GD 1 to PND 21 Dams were dosed via

drinking water throughout
pregnancy until weaning
(PND 21).

Pups (PND 28):
0.93 [jg/dL for control
6.86 [jg/dL for 0.1% Pb

Sex Ratio

BLL = blood lead level; BW = birth weight; d = day(s); F = female; GD = gestational day; M = male; mo = month(s); NR = not reported; Pb = lead; PND = postnatal day; wk = week(s).

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Table 8-4 Epidemiologic studies of Pb exposure and prenatal growth

Reference^and Study stlldy Poplllatlon Exposllre Assessmen,	0llteome	Confcundsrs	Effec, Estates ,„d 95%

Xie etal. (2013)

Shandong Province
China

September 2010 and
December 2011

Cohort

n: 252

Pregnant women aged
18 yr or older, planning
to deliver at the Binhai
hospital, and more than
3 yr of residence in the
Laizhou Bay; exclusion
criteria included
diagnoses of
gestational or
preexisting diabetes,
hypertension, HIV, or
AIDS; GA <28 wk;
known occupational
exposure to heavy
metals; with history of
participation in an
assisted reproduction
program; difficulties
with communication;
and infants with severe
neonatal illnesses

Blood and cord blood

Maternal blood and UCB
were measured by GFAAS.

Age at Measurement:
at delivery (within 3 d before
delivery)

Mean (SD):

Maternal: 3.53 (1.51) pg/dL
UCB: 2.92 (1.58) pg/dL
Median:

Maternal: 3.20 pg/dL
UCB: 2.52 pg/dL
75th:

Maternal: 4.18 pg/dL
UCB: 3.95 pg/dL
Max:

Maternal: 11.91 pg/dL
UCB: 10.60 pg/dL

Prenatal growth: BW,
BL, HC

BW, BL, and HC were
measured by several
trained midwives within
1 hr after birth

Age at outcome:
birth

Multiple linear regression
models were adjusted for
infant sex, maternal
education, maternal age,
GA, pre-pregnancy BMI,
parity, and weight gain
during pregnancy

(3 (95% Cl)b

Maternal blood:

BW (g): -148.99 (-286.33,
-11.66)

BL (cm): -0.46 (-1.25,
0.34)

HC (cm): -0.37 (-0.78,
0.19)

UCB:

BW (g): -99.33 (-217.33,
20.67)

BL (cm): -0.84 (-1.52,
-0.16)

HC (cm): -0.36 (-0.81,
0.03)

Garcfa-Esquinas et al.

(2013)

Madrid
Spain

October 2003 to May
2004

Cohort

BioMadrid Project
n: 112

Father-pregnant
woman-newborn trios
residing in two areas of
the Madrid

Autonomous Region, a
municipal district in the
city of Madrid (urban
area) and a second
zone lying in the
Greater Madrid

Blood and cord blood

Blood collected from both
parents during pregnancy
and UCB was collected at
delivery and measured by
AAS

Age at measurement:
maternal age: >15; birth

Geometric mean (95% Cl)c

Prenatal growth: BW,
length, 1- and 5-min
Apgar scores

Anthropometric data
were measured once,
before breastfeeding
started. Apgar score
was measured on a
scale from 1 to 10, at 1
and 5 min after
delivery. Infants were
evaluated on a scale of

Multivariable linear
regression models were
adjusted for newborn's
sex, GA, maternal age,
maternal cigarette
smoking and sampling
season

(3 (95% Cl)b
UCB:

BW (g): 123 (-37.9, 284)

BL (cm): 0.52 (-0.39, 1.44)

1-min Apgar score: 0.67
(-0.19, 1.16)

5-min Apgar score: 0.29
(-0.04, 0.54)

8-99


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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Metropolitan Belt
(metropolitan area);
women were required
to be aged over 15 yr,
to be expecting a single
pregnancy, and to
intend to deliver their
babies at the public
hospital assigned to
them, lived in the study
area for more than a
year, and did not have
a blood transfusion in
the previous year

Maternal blood: 1.98 (1.816,
2.162) [jg/dL

Paternal blood: 3.30 (3.048,
3.564) [jg/dL
UCB: 1.409 (1.277,
1.555) [jg/dL

Median0:

Maternal blood: 1.898 [jg/dL
Paternal blood: 3.324 [jg/dL
UCB: 1.380 pg/dL
75thc:

Maternal blood: 2.721 pg/dL
Paternal blood: 4.321 pg/dL
UCB: 1.911 pg/dL

0 to 2 according to five
categories (skin color,
muscle tone, reflexes,
respiratory effort, and
heart rate), and the
points from each
category added
together to determine
the total score.

Age at outcome:
birth

Govarts et al.

5 provinces of

Flanders,

Belgium

August 2008-July
2009

Cohort

Flemish human
environmental health
survey (FLEHS II)
n: 248

Women with
uncomplicated live-born
singleton pregnancies,
living in Flanders for at
least 10 yr, ability to fill
in a Dutch
questionnaire, and
giving birth in one out
often randomly
selected maternities

Cord blood

UCB was measured by HR-
ICP-MS

Age at Measurement:
birth

Geometric mean0:
0.864 pg/dL
75thc: 1.138 pg/dL

Prenatal growth: BW Linear regression models (3 (95% CI)b for an increase

BWwas obtained from
the medical records

Age at outcome:
birth

were adjusted for GA,
child's sex, smoking of the
mother during pregnancy,
parity, and maternal pre-
pregnancy BMI

of Z-score of UCB Pb IQR
increase: -37.14 g (-93.64,
19.36)

Tatsuta et al. (2017)

Tohoku
Japan

2000-2003

Tohoku Study of Child Cord blood
Development
n: 489

Singleton pregnancy,
Japanese as the first
language, neonates

UCB was measured by ICP-
MS

Prenatal growth: BW Multiple regression models (3 (p-value)b

BWwas obtained from
medical records

were adjusted for GA,
parity, BMI before
pregnancy,

smoking/drinking habits

All infants: -0.011 g
(0.784)

Male infants: 0.023 g
(0.692)

8-100


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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Cohort

born at term (36-42 wk Age at Measurement:

of gestation) with BW of birth

more than 2400 g, and

no congenital	Median: 1.0 [jg/dL

anomalies or diseases Ma|e jnfants: 1.0 [jg/dL

Female infants: 1.0 [jg/dL

Age at outcome:
birth

during pregnancy, and
fish/seafood intake

Female infants: -0.039 g
(0.513)

95th: 1.7 [jg/dL

Male infants: 1.7 [jg/dL

Female infants: 1.7 [jg/dL

Wang et al. (2017b)

Shanghai
China

September 2008 and
October 2009

Cohort

n: 1,009 mother-infant
pairs

Singleton pregnant
women who had lived
in Shanghai for at least
2 yr, were aged 18 yr or
older, and were
delivering at the
selected hospitals were
recruited. Pregnant
women were excluded
if they had chronic
diseases before
pregnancy, pregnancy
complications, or a
history of occupational
heavy metal exposure.
Infants who had severe
disorders or congenital
malformations at birth
were also excluded.

Cord blood

UCB measured by GFAAS

Age at Measurement:
birth

Geometric mean: 4.07 [jg/dL
(95% CI: 3.98, 4.17)

Prenatal growth: BW,
BL, HC, and the PI

Neonatal
anthropometry,
including BW, BL, and
HC, was performed by
trained delivery room
staff with standardized
equipment, and the
results were recorded.
PI was calculated

Age at outcome:
birth

Multiple linear regression
models; models for BW,
HC, and PI were adjusted
for maternal age, GA,
maternal BMI before
delivery, parity, sex of
baby, monthly household
income per capita, mode
of delivery; models for BL
were maternal age, GA,
maternal BMI before
delivery, parity, sex of
baby, monthly household
income per capita; all
models for female infants
and BL model for male
infants were adjusted for
maternal age, GA,
maternal BMI before
delivery, parity, monthly
household income per
capita; models for male
infants for BW, HC, and PI
were adjusted for maternal
age, GA, maternal BMI
before delivery, parity,
monthly household income

(3 (95% Cl)b
All Infants

BW (g): 50.68 (-69.53,
170.88)

BL (cm): 0.36 (-0.13, 0.86)
HC (cm): -0.39 cm (-0.80,
0.02)

PI (g/cm3): -0.03 (-0.12,
0.07)

Female Infants

BW (g): -139.15 (-317.89,
39.59)

BL (cm): 0.32 (-0.38, 1.03)
HC (cm): -0.13 (-0.71,
0.44)

PI (g/cm3): -0.16 (-0.30,
-0.02)

Male Infants

BW (g): 206.50 g (46.15,
366.86)

BL (cm): 0.35 (-0.35, 1.05)

HC (cm): -0.65 (-1.24,
-0.06)

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RefereDCesignnd Study P°P"'ation

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa







per capita, mode of
delivery

PI (g/cm3): 0.09 (-0.04,
0.21)

Govarts et al.

Belgium

FLEHS I: 2002-2004;
FLEHS II: 2008-2009;
FLEHS III: 2013-
2014; 3xG: 2010-
2015

Cohort

FLEHS I, II and III and
a regional birth cohort
(3xG)

n: 1,579 mother-
newborn pairs: FLEHS
I n = 957, II n = 224, III
n = 273, and 3xG
n = 125

Inclusion criteria were
to be able to fill out a
Dutch questionnaire
and to live at least 5 yr
in the selected study
areas (FLEHS I), at
least 10 yr in Flanders
(FLEHS II), at least 5 yr
in Flanders (FLEHS III),
or living in the
recruitment area (3xG).
Live-born singleton
births

Cord blood

UCB measured by HR-ICP-
MS

Age at Measurement:
birth

Median0:

FLEHS I: 1.42 pg/dL

FLEHS II: 0.83 pg/dL

FLEHS III: 0.61 pg/dL

3xG: 0.61 pg/dL

pooled: 0.97 pg/dL
75thc:

FLEHS I: 2.41 pg/dL
FLEHS II: 1.13 pg/dL
FLEHS III: 0.87 pg/dL
3xG: 0.72 pg/dL
pooled: 1.78 pg/dL

Prenatal growth: BW

BW was recorded
shortly after delivery

Age at outcome:
birth

Multiple linear regression
models were adjusted for
other exposures, GA
(linear and quadratic
terms), sex of the
newborn, maternal age at
delivery, maternal pre-
pregnancy BMI, parity,
smoking during pregnancy
and cohort

(3 (95% Cl)b, interpreted as
the change in mean BW
per interquartile fold
change (the fold change of
the 75th percentile over the
25th percentile in
exposure) in In-Pb: 16.98 g
(-13.14, 47.11) per2.94
interquartile fold change in
In-Pb

Lee et al, (2021)

Dhaka Community

Cord blood

Prenatal growth: BW,

Linear models adjusted for

(3 (95% CI)b, per IQR



Hospital Trust



BL, HC

maternal age, maternal

increase in In-cord blood

Sirajdikhan and Pabna

n: 1088

UCB measured by acid
digestion and ICP-MS



BMI at enrollment, child
sex, GA, household
income, second-hand
smoke, site daily tea

Pb:

Sadar regions



Trained staff measured

Birth Z-scores

Bangladesh



anthropometer at birth;
GA estimated using
<16-wk ultrasound.

BW (g): -0.04 (-0.19, 0.11)

2008-2011



Age at measurement:

(heavy metals) and cord

BL (cm): -0.06 (-0.20,



birth

blood As, Cd, Mn

0.09)

Cohort





Age at outcome: birth

concentrations

HC (cm): 0.08 (-0.06, 0.23)



Geometric mean (Geometric
SD): 3.18 (2.35) pg/dL
Median: 3.07 pg/dL



Untransformed birth size
measurements

8-102


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Reference and Study
Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa



75th: 6.04 pg/dL
Max: 83.5 pg/dL





BW (g): -20.68 (-78.43,
37.08)

BL (cm): -0.23 (-0.61,

0.15)

HC (cm): 0.08 (-0.10, 0.25)

Xu eta!. (2012)

Guiyu and Xiamen
China

2001-2008

Cohort

n: 531 (n = 432 from
Guiyu and n = 99 from
Xiamen)

Women who gave birth
in Guiyu or non-urban
area of Xiamen
between 2001 and
2008

Cord blood

UCB measured by GFAAS

Age at Measurement:
birth

Median:

Guiyu: 10.78 pg/dL

Xiamen: 2.25 [jg/dL
Max:

Guiyu: 47.46 |jg/dL
Xiamen: 7.22 [jg/dL

Prenatal growth: BW,
LBW rate, IUGR rate,
GA

Obtained from birth
records; LBW was
defined as <2500 g

Age at outcome:
birth

Multiple linear and logistic (3 (95% Cl)b
regression models were Mean BW (g): _91 80i

adjusted for maternal age
and infant sex

-75)

Mean GA (wk): 0.57 (0.51,
0.63)

OR (95% Cl)b
LBW: 1.61 (1.37, 1.90)
IUGR: 2.12 (1.68, 2.69)

Al-Saleh et al. (2014) n: 1,578

Al-Kharj
Saudi Arabia

2005-2006

Cohort

Women aged 16-50 yr
who delivered in Al-
Kharj hospital, Saudi
Arabia

Cord blood

UCB measured by AAS

Age at Measurement:
maternal age 16-50; birth

Mean (SD):

UCB: 2.551 (2.592) pg/dL
Median:

UCB: 2.057 pg/dL
75th:

UCB: 2.689 pg/dL
Max:

UCB: 56.511 pg/dL

Prenatal growth: PI

PI was calculated as
BW (kilograms) divided
by birth height (m)
cubed

Age at outcome:
birth

Logistic regression model
was adjusted for maternal
age, parity, mother's third
trimester BMI, urinary
cotinine, geographical
distribution of current
dwelling, newborn
mother's highest
education, total family
income, and GA

OR (95% Cl)b: 0.766
(0.502, 1.167)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Kim et al. (2020)

Guiyu and Haojiang
China

Cross-sectional

e-REACH Study
n: 314

Women 18 yr or older
with a singleton
pregnancy, had lived in
their respective town
for the duration of their
pregnancy, and
consented to
participate in the study.
Women were excluded
if they had a multiple
pregnancy, used
assistive reproductive
technology to become
pregnant, had a history
of psychiatric or thyroid
disorders, or lived
outside of their
respective town for a
cumulative of 3 mo or
more during their
pregnancy

Blood

Maternal blood, collected at
delivery, was measured by
GFAAS

Age at Measurement:
>18 (age at delivery)

Geometric mean:

Guiyu: 6.7 [jg/dL
Haojiang: 3.8 [jg/dL
Max:

Guiyu: 27 [jg/dL
Haojiang: 16 [jg/dL

Prenatal growth: BW,
HC, GA, newborn BMI,
PI

GA was calculated
based on the LMP and
the date of delivery.
Newborn BMI and PI
were calculated using
the recorded BW and
BL.

Age at outcome:
>18 (age at delivery)

Multiple linear and logistic
regression models were
adjusted for maternal age,
maternal education,
maternal occupation,
maternal BMI, gravidity,
ETS, and neonate sex

(3 (95% Cl)b, interpreted as
the difference in BW, HC,
BMI, or PI, per 1 -unit
increase In-Pb maternal
blood

BW (g): 60 (-15, 135)

HC (cm): -0.75 (-1.17,
-0.32)

BMI (kg/m2):

-0.14 (-0.39, 0.11)
PI (kg/m3): -0.62 (-1.13,
-0.11)

OR (95% Cl)b
SGA: 0.69 (0.33, 1.46)

Xu et al. (2022b)

Ushuaia (South,
higher income) and
Salta (North, lower
income)

Argentina

2011-2012

Cross-sectional

EMASAR
n: 696

Women who either
were about to deliver or
had given birth within
the last 48 hr at one of
the two hospitals.
Women had to be
above 18 yr of age.

Blood

Maternal blood measured by
ICP-MS

Age at measurement:
birth

Median0:

Overall: 1.34 pg/dL
Ushuaia: 0.98 [jg/dL
Salta 1.50 pg/dL

Prenatal growth: GA, Linear models adjusted for (3 (95% CI):

BW, BL, HC, LBW

Medical records were
used to obtain
measures at birth.

Age at outcome: birth

maternal age, pre-
pregnancy BMI, parity,
smoking, and education

BW (g): -47.23 (-94.46,
0.004)

BL (cm):

-0.439 (-0.658, -0.219)
HC (cm):

-0.223 (-0.385, -0.061)
GA(wk): 0.18 (0.05, 0.309)

OR (95%CI)

LBW:

T1: Reference
T2: 0.59 (0.10, 3.55)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Geometric mean0:
Overall: 1.393 [jg/dL
Ushuaia: 1.01 [jg/dL
Salta 1.58 pg/dL

T3: 0.53 (0.09, 3.16)

75thc:

Overall: 1.851 pg/dl_
Ushuaia: 1.30 [jg/dL
Salta: 2.09 [jg/dL

Hu etal. (2015)

Beijing, Lanzhou,
Taiyuan, Xiamen
China

June-August 2011
Cohort

n: 81

Mother-infant pairs that
were enrolled from 4
hospitals in 4 cities in
China

Blood and cord blood

Maternal blood (serum) and
UCB (serum) were
measured by ICP-MS

Age at Measurement:
median maternal age:
28.5 yr (range: 18-44); at
birth

Prenatal growth: BW

BWwas obtained from
the medical delivery
records

Age at outcome:
birth

Multivariate linear
regression models were
adjusted for infant gender,
maternal age, gestational
week, and maternal BMI

(3 (95% Cl)b

Maternal serum Pb: -1.7 g
(-9.1, 5.6)

UCB serum Pb: -1.5 g
(-5.2, 8.2)

Median:

Maternal: 23.1 ng/g

UCB: 22.0 ng/g
75th:

Maternal: 33.2 ng/g
UCB: 33.7 ng/g

Yang et al. (2020)

Wuhan
China

2014-2015

Births at Women and
Children Medical and
Healthcare Center of
Wuhan
n: 734

The participants were

Cord blood

UCB (serum) was measured
by ICP-MS

Age at Measurement:
birth

Prenatal growth: BW
(BWGA Z-score)

Midwives immediately
measured BW after
delivery and was
standardized for

Generalized linear
regression models
adjusted for maternal age,
annual household income
levels, pre-pregnancy BMI,
parity, passive smoking
during pregnancy,

P (95% Cl)b, per unit
increase in In-Pb UCB
serum

Continuous: 0.01 (-0.002,

0.05)

Quartiles:

8-105


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Outcome

Confounders

Effect Estimates and 95%
Clsa

Cohort

enrolled at their first
antenatal examination
^gestational 16 wk).
The inclusion criteria
were (1) residence in
Wuhan city; (2) with a
single gestation; (3)
willing to take the
following prenatal care
during pregnancy and
give birth at the study
hospital; (4) willing to
complete

questionnaires and
provide blood samples
from the umbilical cord
at delivery.

Geometric mean: 1.65 |jg/L
Median: 2.71 |jg/L
75th: 4.29 pg/L

Quartilesd

gestational weeks to
construct BWZ.

Age at outcome:
birth

maternal weight gain
during pregnancy, fetal
sex

Q1
Q2
Q3

<25th percentile
25th-50th percentile
50th-75th percentile

Q4: >75th percentile

Q1
Q2
Q3
Q4

Reference
0.11 (-0.09, 0.30)
-0.05 (-0.24, 0.14)
0.05 (-0.14, 0.24)

p for trend: 0.84

Tang et al.

Shengsi Island,

Hangzhou

China

July 2011 to May 2012
Cohort

n: 103

Eligible pregnant
women included those
planning to deliver at
the only hospital,
without apparent
clinical symptoms,
without any maternal
history of illness, and
no poor habits such as
drug use. Eligible
infants were singleton
births and had no
congenital diseases.

Cord blood

UCB (serum) measured by
ICP-MS

Age at Measurement:
birth

Mean (SD)C: 12.841
(28.646) [jg/dL

Median0: 7.620 pg/dL
75thc: 11.580 pg/dL

Tertiles (pg/dl_)c:
T1: <5.633
T2: 5.633-9.197
T3: >9.197

Prenatal growth: BW,
length (height), and HC
and GA

All of these infant
anthropometric
measurements were
collected at birth by
professional healthcare
workers. GAwas
obtained using the
reported date of the
LMP and delivery date.

Age at outcome:
birth

Multivariable linear
regression models were
adjusted for maternal BMI,
maternal age, education
level, newborn gender,
number of abortions,
parity, and pregnancy
weight gain

(3 (95% Cl)b

BW (g): -0.019 (-0.045,
0.006)

BL (cm): 0.29 (-0.50,
-0.09)

HC (cm): -0.22 (-0.44,
-0.00)

GA (wk): -0.21 (-0.44,
0.03)

BW, in g:

T1
T2
T3

Reference
-0.15 (-0.41, 0.11)
-0.05 (-0.30, 0.21)

BL, in cm:

T1
T2
T3

Reference
-0.13 (-0.39, 0.13)
-0.15 (-0.40, 0.11)

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Outcome

Confounders

Effect Estimates and 95%
Clsa

HC, in cm:
T1: Reference
T2: -0.31 (-0.59, -0.02)
T3: -0.13 (-0.40, 0.14)

GA, in wk:

T1
T2
T3

Reference
-0.23 (-0.50, 0.05)
-0.20 (-0.49, 0.08)

Freire et al. (2019)
Spain

2000-2008
Cohort

Environment and
Childhood (INMA)
Project
n: 327

Pregnant women of
general population
resident in each study
area [Ribera d'Ebre,
Menorca, Granada,
Valencia, Sabadell,
Asturias and Gipuzkoa]
and their children.
Criteria for inclusion of
the mothers were: (i) to
be resident in one of
the study areas, (ii) to
be at least 16 yr old,
(iii) to have a singleton
pregnancy, (iv) to not
have followed any
program of assisted
reproduction, (v) to
wish to deliver in the
reference hospital and
(vi) to have no

Other: Placenta

Placenta (including maternal
and fetal sides as well as
central and peripheral parts)
measured with GFAAS
using AAS with Zeeman
background correction

Age at Measurement:
birth

Median: <6.5 ng/g (LOD)
75th: <6.5 ng/g (LOD)

Prenatal growth: BW,
length, HC, LBW, GA,
and SGA

Neonatal
anthropometric
measurements were
obtained by the
attending midwife or
nurse; GA was
calculated as the
number of weeks from
the self-reported LMP
to the end of
pregnancy; LBW was
defined by a BW of
less than 2500 g at
term, newborns were
defined as SGA when
below the 10th
percentile of the
expected weight
according to the
Spanish BW curve
adjusted for GA and
sex

Linear models or logistic
regression models were
adjusted for adjusted for
cohort (random effect),
newborn sex, and co-
exposure to other metals
(As, Hg, Cd, Mn, Cr); BW
and LBW models were
additionally adjusted for
GA, maternal smoking
during pregnancy,
maternal working during
pregnancy, and pre-
pregnancy BMI; BL
models were additionally
adjusted for GA and
maternal smoking during
pregnancy; HC models
were additionally adjusted
for GA, maternal smoking
during pregnancy, pre-
pregnancy BMI, and
cesarean delivery; GA
models were additionally
adjusted for maternal
education level; SGA
models were additionally
adjusted for father's

(3 (95% Cl)b

BW (g): 54.57 (-70.84,
180.0)

BL (cm): -0.26 (-0.97,
0.44)

HC (cm): -0.10 (-0.57,
0.36)

GA (wk): -0.11 (-0.57,
0.36)

OR (95% Cl)b

LBW: 2.94 (0.38, 28.34)

SGA: 1.69 (0.53, 8.82)

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Confounders Effect Esti™*fs and 95%



communication
problems

Age at outcome:
birth

education and maternal
working during pregnancy

ikelson et al. (2019) n: 374

Chattanooga, TN
United States

Cohort

Singleton births of HIV
and hepatitis negative
mothers over 18 yr of
age, with GA greater
than 34 wk, and infants
with no major
morphological or
chromosomal
abnormalities

Other: Placenta

Placenta tissue measured
by ICP-MS

Age at Measurement:
birth

Mean (SD): 37.97
(270.5) pg/kg

Median: 12.03 pg/kg
75th: 23.23 pg/kg
Max: 5073 pg/kg

Prenatal growth: BW

Obtained at birth
records

Age at outcome:
birth

Multivariable regression
models adjusted maternal
pre-pregnancy BMI,
maternal age, GA, race,
infant sex, and smoking
while pregnant

(3 (95% Cl)b: -58.3 g
(-97.9, -18.8)

(3 (95% CI)b, as estimated
change in BW from 25th to
75th percentile: -72.7 g
(-122, -23.4)

iloom et al.

Michigan (4 counties)
and Texas (12
counties)

United States

2005-2009

Cohort

LIFE
n: 235

Potential participants
were identified, using
fishing license
registries or a
commercially available
direct marketing data
base, from 12 counties
in Texas and four in
Michigan, respectively,
with presumed
exposure to persistent
organic pollutants.
Inclusion criteria
comprised a committed
heterosexual
relationship, women
aged 18-40 yr (men
>18), English or
Spanish speaker, no
use of an injectable

Blood

Maternal and paternal blood,
collected before pregnancy
(baseline), were measured
by ICP-MS

Age at Measurement:
>18, maternal mean age:
29.75 (SD: 3.73) yr and
paternal mean age: 31.52
(SD:4.57) yr

Mean (SD):

Maternal: 0.71 (0.30) pg/dL
Paternal: 1.13 (0.63) pg/dL
Median:

Maternal: 0.66 pg/dL

Paternal: 0.98 pg/dL
Max:

Prenatal growth: GA,
BW, BL, HC, PI, and
secondary sex ratio

Women were followed
until delivery when they
completed and
returned birth
announcements that
captured date and sex
of birth, weight and
length, and HC.
Secondary sex ratio is
the ratio of live male to
female births, reflecting
a male excess.

Age at outcome:
birth

Multiple regression models
for continuous outcomes:
effect of maternal
exposure adjusted for
paternal exposure,
maternal age, difference in
maternal and paternal age,
and maternal and paternal
smoking, income, race,
serum lipids (mg/dL), and
creatinine for urine
(mg/dL); effect of paternal
exposure adjusted for
maternal exposure,
paternal age, difference in
maternal and paternal age,
and maternal and paternal
smoking, income, race,
serum lipids (mg/dL), and
creatinine for urine
(mg/dL)

(3 (95% CI):

GA, in days
Maternal Exposure:
T1: Reference
T2: 0.43 (-0.48, 1.35)
T3: 0.14 (-0.81, 1.09)
p for trend: 0.671
Paternal Exposure:

T1
T2
T3

Reference
0.19 (-0.70, 1.08)
0.61 (-0.31, 1.53)

p for trend: 0.416
BW, in kg

Maternal Exposure:

T1: Reference

T2: 81.80 (-79.94,
2238.55)

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Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

contraceptive within
12 mo, and a menstrual
cycle length of 21-42 d.

Maternal: 2.23 [jg/dL
Paternal: 6.43 [jg/dL

Tertiles (|jg/dL):

Maternal Blood Pb
T1: <0.55 (<33rd percentile)
T2: 0.55-0.73 (33rd to 67th
percentile)

T3: >0.73 (>67th percentile)

Paternal Blood Pb

T1: <0.84 (<33rd percentile)

T2: 0.84-1.16 (33rd to 67th
percentile)

T3: >1.16 (>67th percentile)

T3: -34.885 (-197.76,
128.06)

p for trend: 0.202

Paternal Exposure:

T1: Reference

T2: 20.46 (-134.17,
175.09)

T3: 62.91 (-94.73, 220.55)
p for trend: 0.882

BL, in cm

Maternal Exposure:
T1: Reference
T2: 0.43 (-0.48, 1.35)
T3: 0.14 (-0.81, 1.09)
p for trend: 0.671
Paternal Exposure:

T1
T2
T3

Reference
0.19 (-0.70, 1.08)
0.61 (-0.31, 1.53)

p for trend: 0.416

HC, in cm
Maternal Exposure:
T1: Reference
T2: 0.03 (-0.68, 0.74)
T3: -0.33 (-1.07, 0.41)
p for trend: 0.132
Paternal Exposure:

T1
T2
T3

Reference
0.12 (-0.57, 0.80)
-0.03 (-0.72, 0.67)

p for trend: 0.971

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Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

PI, in kg/cm3
Maternal Exposure:
T1: Reference
T2: 0.82 (-7.66, 9.31)
T3: -4.26 (-13.16, 4.64)
p for trend: 0.321
Paternal Exposure:

T1
T2
T3

Reference
-0.22 (-8.50, 8.05)
-5.19 (-13.71, 3.33)

p for trend: 0.150

Rabito et al. (2014)

Shelby County,
Tennessee
United States

2008-2011

Cohort

CANDLE study
n: 98

Healthy pregnant
woman between the
ages of 16 and 40 yr,
carrying a single fetus
with the intent to deliver
the fetus, residence
within Shelby County,
Tennessee, and having
the intent to deliver at
one of three area-
based hospitals

Blood and cord blood

Maternal blood and UCB
were measured by ICP-MS

Age at Measurement:
Maternal age at collection
(second or third trimester or
delivery) (median: 29.50 yr);
birth

Median:

Second trimester:
0.43 [jg/dL

Third trimester: 0.43 |jg/dL
At delivery: 0.50 [jg/dL
Cord blood: 0.37 [jg/dL

Geometric mean (SD):

Second trimester: 0.42
(0.20) [jg/dL

Prenatal growth: BW

BWwas obtained from
medical records

Age at outcome:
birth

Linear regression models
were adjusted for gravidity,
marital status, and GA (in
weeks)

(3 (95% Cl)b, per 0.1 -unit
increase in second
trimester maternal blood
Pb: -43.21 g (-88.6, 2.18)

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Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Third trimester: 0.45
(0.28) [jg/dL

At delivery: 0.50
(0.35) [jg/dL
Cord blood: 0.37
(0.32) [jg/dL

Max:

Second trimester:
1.22 [jg/dL

Third trimester: 2.10 |jg/dL
At delivery: 2.47 [jg/dL
Cord blood: 1.80 [jg/dL

Shih et al. (2021)

United States

January 2009-
September 2010

Cohort

Initial Vanguard Study
of the National
Children's Study

n: 125 (68 males, 57
females)

Mother-infant pairs
enrolled in the National
Children's Study.

Blood

Maternal blood was
measured using dynamic
reaction cell ICP-MS.

Age at measurement:

Maternal age at 6-32 wk of
gestation

Median:

Overall: 0.34 [jg/dL
Male infants: 0.35 [jg/dL
Female infants: 0.33 [jg/dL

Max:

Overall: 2.86 [jg/dL
Male infants: 2.86 [jg/dL
Female infants: 0.85 [jg/dL

Prenatal growth: BL,
HC, BW, GA, and PI

Birth outcomes
measured during
physical examination of
infants at birth; BL (cm)
and HC (cm) were
measured twice, and
the average of the two
readings was used. For
those without
measures at birth,
medical records were
extracted by the
National Children's
Study. GA and BW
were obtained from
medical records.

Age at outcome: birth

Linear regression models
adjusted for maternal age,
race/ethnicity, education,
income, smoking status
during pregnancy, number
of prior livebirths,
continuous BMI, and infant
sex

(3 (95% Cl)b, as expected
change for birth outcomes

GA (wk)

Overall: -0.558 (-2.297,
1.181)

Males: 1.084 (-0.855,
3.024)

Females: -4.335 (-7.365,
-1.305)

BW (g)

Overall: -403.593
(-916.671, 109.485)
Males: 141.814 (-431.7,
715.329)

Females: -1685.349
(-2581.105, -789.592)

BL (cm)

Overall: -0.343 (-2.92,
2.233)

Males: 2.211 (-0.667,
5.089)

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Reference^and Study stlldy Poplllatlon Exposllre Assessmen,	0llteome	Confcundsrs	Effec, Estates ,„d 95%

Females: -6.37 (-10.86,
-1.88)

HC (cm)

Overall: -1.245 (-2.769,
0.279)

Males: 0.292 (-1.397,
1.981)

Females: -4.866 (-7.52,
-2.212)

PI (kg/m3)

Overall: -3.134 (-6.698,
0.429)

Males: -2.377 (-6.486,
1.731)

Females: -4.733 (-11.191,
1.725)

Woods etal. (2017)

Cincinnati, Ohio
United States

2003-2006

Cohort

HOME Study
n: 272

Women were recruited
between 13 and 19 wk
of pregnancy from
prenatal clinics and
were >18 yr old,
<19 wk gestation at the
time of enrollment, and
living in a residence
built before 1987

Blood

Maternal blood was
measured by sensitive and
specific liquid or gas
chromatography mass
spectrometry

Age at measurement:
maternal age at 16-26 wk
gestation

Geometric mean (geometric
SD): 0.7 (1.4) [jg/dL

Median: 0.7 [jg/dL
75th: 0.8 pg/dL

Prenatal growth: BW

BW was abstracted
from birth records

Age at outcome:
birth

Bayesian hierarchical
linear models were
adjusted for maternal race,
age at delivery, infant sex,
maternal education,
tobacco exposure,
household annual income,
employment, maternal
insurance status, marital
status, pre-natal vitamin
use, and maternal BMI;
sensitivity analysis
included GA

Posterior mean (95%
credible interval)15, as the
difference in BW per 10-
fold increase in maternal
blood Pb: -44.8 g (-110,
21.7)

Taylor et al. (2016) ALSPAC	Blood	Prenatal growth: BW, Multivariable fractional (3 (95% CI)

n: 4,190	HC, crown-to-heel polynomials and modeled

Bristol, UK	length	adjusted for maternal

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Confounders

Effect Estimates and 95%
Clsa

April 1991-December
1992

Cohort

All pregnant women in
the former Avon Health
Authority with an
expected delivery date
between April 1,1991,
and December 31,
1992, were eligible for
the study; 14,541
pregnant women were
initially enrolled,
resulting in a cohort of
14,062 live births

Maternal blood was
measured by ICP-MS

Age at measurement:
maternal age at
measurement (median GA
of sampling: 11 wk)

Median: 3.40 [jg/dL
75th: 4.33 pg/dL
Max: 19.41 pg/dL

HC and CHL were
measured by trained
study staff where the
mother gave
permission or if these
data were missing, the
values were extracted
from the medical
records by trained
study staff. BW was
derived from obstetric
data and from central
birth notification data:
where values
disagreed by <100 g
then the lowest value
was accepted; if the
values disagreed by
>100 g then the value
was coded as missing.

Age at outcome:
birth

educational attainment,
smoking, GA (centered at
40 wk), maternal height
and pre-pregnancy weight,
and sex of the infant

BW (g): -9.93 (-20.27,
0.41)

HC (cm): -0.03 (-0.06,
0.00)

CHL (cm): -0.05 (-0.10,
0.00)

Garcfa-Esquinas et al.

Madrid
Spain

October 2003-May
2004

Cohort

BioMadrid Project
n: 97

Women were required
to be aged over 15 yr,
to be expecting a single
pregnancy, intend to
deliver their babies at
the public hospital
assigned to them, lived
in the study area
(Madrid Autonomous
Region) for more than a
year, and did not have

Blood and cord blood

Blood, from both parents,
and UCB measured by AAS
with a transversely heated
graphite atomizer furnace
assembly and longitudinal
Zeeman-effect background
correction

Age at measurement:
maternal and paternal age
at median gestational week
was 33.9 (IQR 31.6-35.7)
and at birth

Prenatal growth: GA,
BW, BL, AD, or CD

GA, BW, BL, AD, or
CD was collected at
delivery

Age at outcome:
birth

Multivariable linear
regression models were
adjusted for sampling
maternal age, maternal
tobacco smoke, area
(metropolitan/urban), and
in non-stratified models,
newborn's sex

(3 (95% CI)b, as mean
difference per two-fold
increase in BLL

Maternal blood Pb

BW (g): 62.4 (-73.1, 197.8)

BL (cm): 0.17 (-0.56, 0.91)

AD (cm): 0.31 (-0.52, 1.15)

CD (cm): 0.15 (-0.21, 0.51)

GA (wk): 0.02 (-0.44, 0.47)

Paternal blood Pb
BW (g): -110.8 (-235.6,
6.0)

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Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

a blood transfusion in



BL (cm): -0.44 (-1.12,

the previous year

Geometric mean0:

0.23)



Maternal: 1.83 [jg/dL

AD (cm): -0.81 (-1.64,



Paternal: 3.17 [jg/dL

-0.00)



UCB: 0.45 pg/dL

CD (cm): -0.32 (-0.65,



0.00)





GA(wk): -0.17 (-0.59,





0.26)





UCB Pb





BW (g): 80.0 (-36.8, 196.6)





BL (cm): 0.30 (-0.33, 0.93)





AD (cm): 0.56 (-0.12, 1.24)





CD (cm): -0.16 (-0.47,





0.15)





GA(wk): -0.04 (-0.44,





0.35)





Male Infants





Maternal blood Pb





BW (g): 62.6 (-145.2,





270.4)





BL (cm): 0.29 (-0.83, 1.41)





AD (cm): 1.10 (-0.25, 2.45)





CD (cm): -0.16 (-0.47,





0.15)





GA(wk): 0.11 (-0.58, 0.81)





Paternal blood Pb





BW (g): -93.5 (-269.6,





82.5)





BL (cm): 0.13 (-0.81, 1.06)





AD (cm): -0.64 (-1.89,





0.61)

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Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

CD (cm): -0.11 (-0.57,
0.35)

GA(wk): 0.06 (-0.53, 0.65)

UCB Pb

BW (g): 80.0 (-66.0, 226.0)
BL (cm): 0.66 (-0.11, 1.44)
AD: 0.76 cm (-0.16, 1.67)
CD: -0.11 cm (-0.37, 0.39)
GA: 0.06 wk (-0.53, 0.65)

Female Infants

Maternal blood Pb

BW (g): 62.2 (-128.0,
252.4)

BL (cm): 0.08 (-0.95, 1.10)
AD (cm): -0.21 (-1.30,
0.88)

CD (cm): -0.05 (-0.55,
0.46)

GA (wk): -0.06 (-0.70,
0.57)

Paternal blood Pb

BW (g): -129.4 (-312.3,
53.4)

BL (cm): -1.06 (-2.03,
-0.08)

AD (cm): -1.94 cm (-2.06,
0.18)

CD (cm): -0.55 (-1.03,
-0.07)

GA (wk): -0.41 (-1.02,
0.21)

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Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

UCB Pb

BW (g): 80.0 (-115.7,
275.7)

BL (cm): -0.37 (-1.41,

0.67)

AD (cm): 0.31 (-0.73, 1.35)

CD (cm): -0.47 (-0.98,
0.05)

GA(wk): -0.13 (-0.79,
0.65)

Daniali et al. (2023)
Isfahan, Iran
2019-2020
Cohort

Prospective
Epidemiologic
Research Studies in
Iran - Isfahan Center

n: 263

Pregnant Iranian
women who have lived
in Isfahan for at

least 1 yr, and did not
have any history of
infertility, those
in the first trimester of
pregnancy, and those
who intended to give
birth in hospitals of
Isfahan city. All
participants with major
risks of SGA and IUGR
such

as serious medical
complications
(hypertension or
diabetes or kidney
disease), cerclage until
24 wk of pregnancy,

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
maternal age at first
trimester (mean maternal
age 29.94 yr)

Geometric mean ± SD:
2.534 ± 0.205 pg/dL
Median: 2.786 pg/dL
25th: 1.741 pg/dL
75th: 4.01 pg/dL

Prenatal growth: BW,
HC, BL

Standardized neonatal
anthropometric
measurements were
obtained by trained
midwives using
calibrated instruments.

Age at outcome: birth

Infant sex, and maternal
age, BMI at enrollment
(12-14 wk gestation),
income, secondhand
smoke exposure, parity,
and education.

B (95%CI)

BW (g): -0.057 (-0.099,
-0.014)

BL (cm): 0.01 (-0.034,
0.054)

HC (cm): -0.036 (-0.076,
0.004)

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Effect Estimates and 95%
Clsa

history of stillbirth or
preterm labor, multiple
pregnancies, or
abnormal sonographic
evidence were
excluded from the
study.

Taylor et al.

Bristol
UK

April 1991-December
1992

Cohort

ALSPAC
n: 4,285

All pregnant women in
the former Avon Health
Authority with an
expected delivery date
between April 1, 1991,
and December 31,
1992, were eligible for
the study

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
maternal age at
measurement (median GA
of sampling: 11 wk)

Mean (SD): 3.67
(1.47) [jg/dL

Geometric mean: 3.43 [jg/dL

Median: 3.42 [jg/dL
Max: 19.14 pg/dL

Prenatal growth: BW,
HC, CHL, and LBW

BW, HC, and CHL
were measured by
trained staff or
extracted from medical
records; LBW was
<2500 g

Age at outcome:
birth

Linear regression models
were adjusted for maternal
height, maternal pre-
pregnancy weight,
maternal educational
attainment, parity, number
of cigarettes per day, sex
of baby, GA at delivery or
death; logistic regression
models for LBW were
adjusted for maternal
height, maternal pre-
pregnancy weight,
maternal educational
attainment, parity, number
of cigarettes per day, sex
of baby and GA at delivery
or death

(3 (95% CI)

BW (g): -1.62 (-2.909,
-0.331)

HC (cm): -0.005 (-0.043,
0.033)

CHL (cm): -0.006 (-0.013,
0.001)

OR (95% CI)

LBW: 1.37 (0.86, 2.18)

Hu et al. (2021)
Canada
2008-2011
Cohort

MIREC
n: 1857

Women from the
MIREC cohort who
delivered singleton live
births, had complete
sociodemographic
information, and
provided biological
samples during the first
trimester of the
pregnancy.

Blood

Maternal blood was
measured by ICP-MS

Age at Measurement:
Maternal age during first
trimester

Geometric mean0:
0.62 [jg/dL

Prenatal growth: BW

Infant BW (g)
abstracted from
medical records and
examined continuously.

Age at outcome: birth

Maternal age, race,
education, pre-pregnancy
BMI, smoking status,
parity, infant sex, cubic-
spline GA; multi-pollutant
model was also adjusted
for As, Cd, Hg, and Mn

j3 (95% CI), as two-fold
increase in Pb blood

Single pollutant model:
-82.22 g (-145.46, -18

.97)

Multi-pollutant model:
-75.89 g (-141.24, -10.54)

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Effect Estimates and 95%
Clsa

Median0: 0.60 [jg/dL
75thc: 0.85 pg/dL

Goto et al. (2021)
Japan

January 2011-March
2014

Cohort

JECS
n: 16,423

Pregnant women living
in the study area and
understanding ofthe
Japanese language.
Participants were
excluded: if they did not
meet the Pb
measurement quality
control criteria
(n = 2,002); if mothers
who: were lost to
follow-up; had severe
maternal conditions
preceding pregnancy,
such as chronic
hypertension,
pregestational diabetes
or cardiac disease,
during pregnancy; or
had pregnancies
ending in abortions or
stillbirths (n = 1,209); if
infants had

chromosomal or major
congenital anomalies
(n = 263) or multiple
births (n =283)

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
maternal age at second or
third trimester (mean age at
delivery: 31 ± 5.0 yr)

Mean: 0.69 pg/dL
Median: 0.63 pg/dL
75th: 0.78 pg/dL
Max: 7.4 pg/dL

Prenatal growth: BW,
SGA, and LBW

BWwas the primary
outcome.

Anthropometric data
were measured by
trained delivery room
staff. Gestational
dating was performed
from the first accurate
ultrasound examination
during the first
trimester. SGA was
defined as a BW below
the 10th percentile of
the national BWs
reported in the
Japanese standard
growth chart, which
also considers GA,
infant sex, and
maternal parity. LBW
was defined as a BW
below 2500 g,
regardless of GA.

Age at outcome:
birth

Multivariable linear
regression models were
adjusted for maternal age
at birth, BMI before
pregnancy, weight gain
during pregnancy,
maternal educational
background, a history of
preterm birth, alcohol
consumption during
pregnancy, smoking habit
during pregnancy, and
parity

j3 (95% CI), per 0.1 pg/dL
increase in maternal blood
Pb

BW (g): -54 (-74.5, -33.5)
HC (cm): -0.10 (-0.05,
-0.15)

BL (cm): -0.20 (-0.30,
-0.10)

GA (days): 0.20 d (-0.35,
0.75)

(3 (95% CI), per doubling
increment in maternal
blood Pb

BW (g): -86.595 (-112.16,
-61.03)

HC (cm): -0.152 (-0.25,
-0.054)

BL (cm): -0.326 (-0.468,
-0.185)

GA (days): 0.087 (-0.566,
0.74)

OR (95% CI), per 0.1 pg/dL
increase in maternal BLL

SGA: 1.34 (1.16, 1.55)

LBW: 1.34 (1.16, 1.55)

OR (95% CI), per doubling
increment in maternal
blood Pb

SGA: 1.952 (1.526, 2.498)
LBW: 1.34 (1.16, 1.55)

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Rodosthenous et al.
(2017)

Mexico City
Mexico

2007-2011

Cohort

PROGRESS
n: 944

Inclusion criteria:
singleton pregnancy,
GA <20 wk, maternal
age of >18 yr,
expectation to live in
Mexico City for the
following 3 yr, and have
access to a telephone;
exclusion criteria:
chronic medical
conditions such as
heart or kidney
disease; use of steroids
or anti-epilepsy drugs;
drug addiction; and
daily consumption of
alcoholic beverages
due to its association
with adverse fetal
outcomes

Blood

Maternal blood measured by
ICP-QQQ

Age at measurement:

maternal age at -20 wk
gestation

Mean (SD): 3.7 (2.7) pg/dL

Quartile Mean (SD) (pg/dL):

Q1: 1.4 (0.3)

Q2: 2.4 (0.2)

Q3: 3.6 (0.5)

Q4: 7.3 (2.8)

Median: 2.8 pg/dL

75th: 4.5 pg/dL
Max: 22.9 pg/dL

Quartiles (pg/dL)

Q1: <1.93

Prenatal growth:

BWGA, SGA

Infants with a BWGA Z-
score <10th percentile
as SGA

Age at outcome:
birth

Q2
Q3
Q4

1.93-2.79
2.80-4.53
>4.53

Multivariable linear
regression models were
adjusted for maternal age,
BMI, SES, hemoglobin
levels, and infant sex

Quantile regression
models were adjusted for
maternal age, BMI, SES,
hemoglobin levels, and
infant sex

Multivariable logistic
regression models were
adjusted for maternal age,
BMI, SES, hemoglobin
levels, and infant sex

(3 (95% Cl)b, as difference
in BWGA Z-score per log2
increase in maternal BLL:
-0.06 (-0.013, 0.03)

(3 (95% Cl)b, as the BWGA
Z-score per log2 increase in
maternal BLL

QL 0.05: -0.08 (-0.19,
0.03)

QL 0.10: -0.13 (-0.25,
-0.004)

QL 0.15: -0.11 (-0.22,
-0.002)

QL 0.20: -0.12 (-0.20,
-0.03)

QL 0.25: -0.10 (-0.19,
-0.02)

QL 0.30: -0.11 (-0.18,
-0.04)

QL 0.35: -0.04 (-0.12,
0.04)

QL 0.40: -0.06 (-0.14,
0.03)

QL 0.45: -0.05 (-0.13,
0.04)

QL 0.50: -0.07 (-0.16,
0.01)

QL 0.55: -0.07 (-0.16,
0.01)

QL 0.60: -0.07 (-0.15,
0.01)

QL 0.65: -0.04 (-0.12,
0.04)

QL 0.70: -0.04 (-0.12,
0.03)

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Effect Estimates and 95%
Clsa

QL 0.75: -0.01 (-0.08,
0.06)

QL 0.80: -0.02 (-0.1, 0.06)

QL 0.85: -0.06 (-0.16,
0.04)

QL 0.90: -0.06 (-0.16,
0.02)

QL 0.95: -0.02 (-0.13,
0.09)

OR (95% CI) for SGA:
Q1: Reference
Q2: 1.30 (0.79, 2.15)
Q3: 1.15 (0.92, 1.45)
Q4: 1.09 (1.00, 1.18)
p for trend: 0.06

Ashrap et al. (2020)

Puerto Rico

2010-2017

Cohort

PROTECT
n: 731

Participants were
recruited at
approximately
14 ± 2 wk of gestation
at seven prenatal
clinics and hospitals
throughout Northern
Puerto Rico and
followed until birth;
maternal age between
18 and 40 yr; residence
inside of the Northern
Karst aquifer region;
disuse of oral
contraceptives within
the 3 mo prior to
pregnancy; disuse of
IVF to become

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
18-40 (collection between
18 and 26 wk of gestation)

Geometric mean (SD):

Preterm births: 0.39
(1.6) pg/dL

Term births: 0.32 (1.5) pg/dL
Median:

Preterm births: 0.36 [jg/dL
Term births: 0.32 [jg/dL

Prenatal growth: GA,
SGA, LGA, BWZ

All the birth outcome
data were extracted
from medical records.
GA was calculated;
BWZ was defined as
the number of SDs by
which a BW is above or
below the mean; SGA
births were defined as
below the 10th
percentile of BWZs;
LGA births were
defined as above the
90th percentile of
BWZs

Logistic regression models
were adjusted for maternal
age, maternal education
level, pre-pregnancy BMI,
and exposure to second-
hand smoking

(3 (95% Cl)b, per change
per IQR increase in
maternal blood In-Pb

GA (days): -1.
-0.5)

Tertilesd:
GA (days):

(-3.1,

T1
T2
T3

Reference
-0.2 (-2.9, 2.4)
-2.9 (-5.5, -0.2)

BWZ:

T1
T2
T3

Reference
-0.12 (-0.32, 0.07)
0.09 (-0.11, 0.29)

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pregnant; and free of
any major medical or
obstetrical

complications, including
pre-existing diabetes.
Each woman
participated in a total of
up to three study visits
(18 ± 2 wk, 22 ± 2 wk,
and 26 ± 2 wk of
gestation).

Age at outcome:
birth

OR (95% Cl)b, per change
per IQR increase in
maternal blood In-Pb
SGA: 0.91 (0.69, 1.2)

Tertilesd:

SGA

T1: Reference
T2: 1.58 (0.88, 2.83)
T3: 0.62 (0.30, 1.26)
LGA

T1: Reference
T2: 1.13 (0.63, 2.03)
T3: 0.74 (0.40, 1.40)

Thomas et al. (2015)

Canada

2008-2011
Cohort

MIREC Study
n: 1,835

Pregnant women were
recruited in the first
trimester of pregnancy
from 10 study sites
across Canada.
Exclusion criteria
included: inability to
communicate and
consent in either
French or English,
>14 wk gestation at the
time of recruitment,
<18 yr of age,
diagnosed with a fetal
anomaly or a history of
major chronic disease.
Excluded from the
analysis were: 18
women who withdrew

Blood

Maternal blood, collected
during the first and third
trimesters of pregnancy,
was measured by ICP-MS

Age at Measurement:
Maternal age at first and
third trimesters

Median: 0.59 [jg/dL
75th: 0.81 pg/dL
Max: 4.04 pg/dL

Tertiles (pg/dL):

T1
T2
T3

<0.52

0.52-1.04

>1.04

Prenatal growth: SGA

SGA births were
identified as those
weighing less than the
10th percentile for a
reference population
based on the same
completed week of
gestation and infant
sex

Age at outcome:
birth

Log binomial multivariate
regression models
estimated RR and
adjusted for smoking and
parity

RR (95% CI):
T1: Reference
T2: 1.33 (0.88, 1.99)
T3: 1.19 (0.65, 2.18)

RR (95% CI) for GSTP1

A114V

CC

Pb <0.08 pg/dL: Reference
Pb >0.08 pg/dL: 0.90 (0.57,
1.41)

TC + tT

Pb <0.08 pg/dL: Reference

Pb >0.08 pg/dL: 2.25 (0.95,
5.16)

p for interaction: 0.06

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Effect Estimates and 95%
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during the study, 51
women who gave birth
to multiples, 9
stillbirths, 32
spontaneous abortions,
13 therapeutic
abortions, 28 with no
metal exposure data,
and 15 with no infant
sex, weight, or GA
recorded

RR (95% CI) for GSTP1

1105V

AA

Pb <0.08 [jg/dL: Reference
Pb >0.08 [jg/dL: 1.22 (0.69,
2.15)

AG + GG

Pb <0.08 [jg/dL: Reference
Pb >0.08 [jg/dL: 0.95 (0.54,
1.66)

p for interaction: 0.53

RR (95% CI) for GSTOI

A104A

CC

Pb <0.08 [jg/dL: Reference
Pb >0.08 [jg/dL: 0.94 (0.52,
1.69)

CA + AA

Pb <0.08 [jg/dL: Reference

Pb >0.08 [jg/dL: 1.20 (0.70,
2.06)

p for interaction: 0.54

Ashrap et al. (2021)

Puerto Rico

2011-2017

Cohort

PROTECT
n = 682

Participants were
recruited at
approximately
14 ± 2 wk of gestation
at seven prenatal
clinics and hospitals
throughout Northern
Puerto Rico and

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
18-40 (collection between
18 and 26 wk of gestation)

Prenatal growth: GA,
BWZ, small for
gestation, large for
gestation

Birth outcomes were
extracted from medical
records. Psychosocial
status was evaluated
using four
questionnaires

Linear and logistic
regression models were
adjusted for maternal age,
maternal education, pre-
pregnancy BMI, second-
hard smoke exposure

(3 (95% CI)b, per IQR
increase in in maternal
blood In-Pb
GA, change in days

Good Psychosocial Status:
-1.9 (-3.2, -0.6)

Poor Psychosocial Status:
-1.3 (-4.0, 1.5)

BWZ, change in Z-score

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followed until birth;
maternal age between
18 and 40 yr; residence
inside of the Northern
Karst aquifer region;
disuse of oral
contraceptives within
the 3 mo prior to
pregnancy; disuse of
IVF to become
pregnant; and free of
any major medical or
obstetrical

complications, including
pre-existing diabetes.
Each woman
participated in a total of
up to three study visits
(18 ± 2 wk, 22 ± 2 wk,
and 26 ± 2 wk of
gestation)

Geometric mean: 3.1 [jg/dL Age at outcome: birth

Median: 3.1 [jg/dL

75th: 4.1 [jg/dL

95th: 6.5 pg/dL

Max: 15.1 pg/dL

Good Psychosocial Status:
0.1 (0.0, 0.2)

Poor Psychosocial Status:
-0.1 (-0.3, 0.2)

OR (95% CI)b, per IQR
increase in in maternal
blood In-Pb
SGA

Good Psychosocial Status
0.86 (0.65, 1.14)

Poor Psychosocial Status:
1.49 (0.67, 3.33)

LGA

Good Psychosocial Status
0.89 (0.64, 1.23)

Poor Psychosocial Status:
1.10 (0.57, 2.10)

Gustin et al. (2020)

Norrbotten County
Sweden

2015-2018

Cohort

NICE
n: 589

The cohort was
established in the
catchment area of
Sunderby hospital in
Norrbotten county,
Sweden. At the routine
ultrasound in
gestational week 17-
18, parents who were
interested in
participation were given
more information and
an informed consent to
sign at home and send
back. To be included in
the study, families had

Blood

Maternal blood (erythrocyte)
was measured by ICP-MS

Age at measurement:
Maternal age at gestational
week 24-36 (mean: 31 yr,
range 19-45 yr)

Mean: 14 pg/kg

Median: 11 pg/kg
Max: 148 pg/kg

Prenatal growth: BW,
BL, and HC

Information on the
infants' weight (g),
length (cm), and HC
(cm) at birth was
collected from the
hospital records at
Sunderby hospital.

Age at outcome:
birth

Multivariable-adjusted
linear and spline
regression models were
adjusted for maternal age,
early-pregnancy BMI,
parity, education, pre-
pregnancy smoking, pre-
pregnancy snuff or non-
smoking tobacco use, pre-
pregnancy alcohol
consumption, and
marital/cohabitant status;
infant sex and GA at birth
(in days); models were
also mutually adjusted for
other maternal metals (Cd
and Hg)

(3 (95% CI)b:

BW (g): -13 (-66, 41)
p for interaction with infant
sex: 0.88

BL (cm): -0.080 (-0.31,
0.15)

p for interaction with infant
sex: 0.43
HC (cm):

Less than median: 0.059
(-0.22, 0.34)

p for interaction with infant
sex: 0.84

Greater than median:
-0.24 (-0.53, 0.056)

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Effect Estimates and 95%
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to be residents in
Norrbotten county and
be able to

communicate in written
and spoken Swedish.

p for interaction with infant
sex: 0.23

Mutually adjusted for other
maternal metals
BW (g): -0.0091 (-0.077,
0.058)

BL (cm): -0.0078 (-0.079,
0.064)

HC (cm):

Less than median: 0.018
(-0.058, 0.094)

Greater than median:
-0.050 (-0.13, 0.0026)

Rahman et al. (2021) Project Viva
n: 1391

Massachusetts

United States

1999-2002

Cohort

Women were recruited
at prenatal care visits at
eight urban and
suburban practices of a
multi-specialty group
practice in eastern
Massachusetts.
Exclusion criteria
included multiple
gestation, inability to
answer questions in
English, GA >22 wk at
recruitment and plans
to move away from the
study area before
delivery.

Blood

Maternal blood (erythrocyte)
was measured by ICP-MS.

Age at measurement:
maternal age at collection
(mean 11.3 ± 2.8 wk
gestation); mean maternal
age

(SD): 32.3 (4.7) yr

Geometric mean: 17.99 ng/g
Median: 17.7 ng/g
75th: 23.6 ng/g

Prenatal growth: BW,
BL, HC, GA

GA from reported last
menstrual period, BW,
BL, and HC from
medical records

Age at outcome: birth

Multivariable linear
regression models were
adjusted for maternal age,
education, pre-pregnancy
BMI, parity, smoking
status, race/ethnicity,
household income, infant
sex, and GA at delivery
(except when GA is an
outcome)

(3 (95% CI)b, per IQR
(10.1 ng/g) increase:

BW (g):

Full Cohort: -33.9 (-65.3,
-2.5)

Males: -32.5 (-77.4, 12.5)

Females: -34.6 (-77.2,
8.1)

BL (cm):

Full Cohort: -0.10 (-0.29,
-0.09)

Males: -0.08 (-0.35, 0.19)

Females: -0.13 (-0.39,
0.13)

HC (cm)

Full Cohort: -0.07 (-0.17,
0.04)

Males: -0.14 (-0.29, 0.02)

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Effect Estimates and 95%
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Females: 0.00 (-0.15,
0.15)

GA (wk)

Full Cohort: 0.03 (-0.10,
0.16)

Males: 0.12 (-0.07, 0.30)

Females: -0.04 (-0.22,
0.14)

(95% Cl)b, per IQR
(10.1 ng/g) increase, when
As, Cd, Mn, Zn, and Hg
were fixed at the 25th
percentile:

BW (g):

Full Cohort: -0.05 (-0.11,
0.02)

Males: -0.03 (-0.12, 0.06)

Females: -0.06 (-0.14,
0.02)

BL (cm):

Full Cohort: -0.06 (-0.14,
0.03)

Males: -0.04 (-0.16, 0.07)

Females: -0.04 (-0.14,
0.07)

HC (cm)

Full Cohort: -0.08 (-0.18,
0.02)

Males: -0.08 (-0.26, 0.09)
Females: -0.04 (-0.14,
0.07)

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Effect Estimates and 95%
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GA (wk)

Full Cohort: 0.02 (-0.06,
0.10)

Males: 0.05 (-0.06, 0.16)

Females: -0.01 (-0.12,
0.100)

(3 (95% CI)b, per IQR
(10.1 ng/g) increase, when
As, Cd, Mn, Zn, and Hg
were fixed at the 50th
percentile:

BW (g):

Full Cohort: -0.04 (-0.10,
0.01)

Males: -0.03 (-0.11, 0.05)
Females: -0.05 (-0.13,
0.02)

BL (cm):

Full Cohort: -0.05 (-0.13,
0.04)

Males: -0.04 (-0.15, 0.07)

Females: -0.03 (-0.14,
0.07)

HC (cm)

Full Cohort: -0.06 (-0.15,
0.03)

Males: -0.06 (-0.21, 0.09)
Females: -0.03 (-0.14,
0.07)

GA (wk)

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Effect Estimates and 95%
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Full Cohort: 0.01 (-0.06,
0.08)

Males: 0.04 (-0.06, 0.14)

Females: -0.02 (-0.12,
0.08)

(3 (95% CI)b, per IQR
(10.1 ng/g) increase, when
As, Cd, Mn, Zn, and Hg
were fixed at the 75th
percentile:

BW (g):

Full Cohort: -0.04 (-0.11,
0.02)

Males: -0.03 (-0.12, 0.06)
Females: -0.05 (-0.13,
0.03)

BL (cm):

Full Cohort: -0.04 (-0.12,
0.05)

Males: -0.04 (-0.15, 0.08)
Females: -0.03 (-0.14,
0.08)

HC (cm)

Full Cohort: -0.03 (-0.13,
0.08)

Males: -0.05 (-0.22, 0.13)

Females: -0.03 (-0.14,
0.08)

GA (wk)

Full Cohort: -0.01 (-0.09,
0.07)

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Males: 0.03 (-0.08, 0.14)









Females: -0.03 (-0.14,









0.08)

Wang et al. (2017a)
China

2009

Cohort

C-ABCS
n: 3,125

Pregnant women with
singleton, live births

Blood

Maternal blood (serum) was
detected by GFAAS

Age at measurement:
maternal age at collection
(first trimester, median:
11 wk) and second trimester
(median: 16 wk) (mean age:
27.5 yr)

Mean:

Overall: 1.50 [jg/dL

First trimester: 1.52 [jg/dL

Second trimester:

1.49 [jg/dL

Median:

Overall: 1.43 pg/dL
First trimester: 1.43 |jg/dL
Second trimester:

1.43 [jg/dL
Max:

Overall: 5.46 [jg/dL

First trimester: 5.16 [jg/dL

Second trimester:

5.46 [jg/dL

Tertiles (|jg/dL):

Low: <1.18
Medium: 1.18-1.70

Prenatal growth: SGA,
BW, BL, HC, and CC

SGA was defined as
live-born infants with
BW below 10th
percentile for the
babies of the same GA
according to a global
reference; BW, BL, HC,
and CC were
measured at birth

Age at outcome:
birth

Multivariate linear and
logistic regression models
were adjusted for pre-
pregnancy BMI, maternal
age, gravidity, monthly
income, parity, and time of
serum collection

(3 (95% Cl)b
Maternal serum during
pregnancy

BW (g): -2.74 (-5.17,
-0.31)

BL (cm): -0.013 (-0.026,
0.001)

HC (cm): -0.008 (-0.019,
0.004)

CC (cm): -0.008 (-0.018,
-0.002)

First trimester maternal
serum

BW: -4.40 g (-8.22, -0.58)

BL: -0.022 cm (-0.048,
0.005)

HC: -0.007 cm (-0.022,
0.007)

CC: -0.015 cm (-0.030,

<0)

Second trimester maternal
serum

BW (g): -1.64 (-4.80,
-0.58)

BL (cm): -0.006 (-0.020,
0.009)

HC (cm): -0.008 (-0.024,
0.008)

CC (cm): -0.002 (-0.016,
-0.011)

OR (95% CI)

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RefereDCesignnd	Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

High: >1.71

SGA

All Infants
Low: Reference
Medium: 1.45 (1.04, 2.02)
High: 1.69 (1.22, 2.34)
Males

Low: Reference
Medium: 1.44 (0.83, 2.50)
High: 1.75 (1.03, 2.99)
Females
Low: Reference
Medium: 1.51 (0.99, 2.31)
High: 1.68 (1.12, 2.54)
First trimester maternal
serum

Low: Reference

Medium: 1.19 (0.65, 2.19)

High: 2.13 (1.24, 3.38)

Second trimester maternal
serum

Low: Reference
Medium: 1.57 (1.05, 2.34)
High: 1.48 (0.98, 2.21)

Cassidy-Bushrow et

al. (2019)

Wayne County, Ml
United States

September 2003 and
December 2007
(December 2011 and
January 2015)

WHEALS
n: 145

Pregnant women were
in their second
trimester or later, were
aged 21-49 yr, and
were living in a
predefined geographic
area in Wayne and
Oakland counties that

Teeth

Teeth, representing second
and third trimester exposure,
measured by LA-ICP-MS

Mean (SD)6:

Second trimester: 0.04
(0.03) |jg/g

Prenatal growth: BWZ Linear regression models (3 (95% CI)b

and GA

BWZ and GA obtained
from prenatal and birth
records

Age at outcome:
birth

adjusted for batch, tooth
attrition, tooth type, race,
urban, ETS, anemic,
maternal age, and year
house built; the effect of
time is the difference in
effect estimates from the
second and third
trimesters

BWZ

Second trimester: -0.15
(-0.35, 0.05)

Third trimester: -0.06
(-0.24, 0.12)

Effect of Time: -0.31
(-0.90, 0.28)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Cohort

included the city of
Detroit as well as the
suburban areas
immediately
surrounding the city

Third trimester: 0.05
(0.04) |jg/g

Boys

Second trimester: -0.20
(-0.47, 0.07)

Third trimester: -0.04
(-0.31, 0.23)

Girls

Second trimester: -0.12
(-0.39, 0.15)

Third trimester: -0.06
(-0.33, 0.21)

GA (wk)

Second trimester: 0.08
(-0.19, 0.35)

Third trimester: 0.14
(-0.11, 0.39)

Effect of time: -0.22
(-1.08, 0.64)

Boys

Second trimester: 0.08
(-0.41, 0.57)

Third trimester: 0.01
(-0.44, 0.46)

Girls

Second trimester: 0.12
(-0.21, 0.45)

Third trimester: 0.27
(-0.06, 0.60)

CCG MSA	Births where the mother's Prenatal growth: BW, Difference-in-difference (3 (95% CI)b, as estimated

n' 147 673 live births in residential address was LBW, and SGA	models were used to	average treatment effect of

North Carolina	the CCG MSA'	within 4,000 meters of CMS	compare birth outcomes in treatment group

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

United States

2004-2009

Quasi-experimental

Treatment group n:
1,138; Control group n:
13,398

Exogenous variation in
Pb exposure resulting
from NASCAR's
deleading of racing fuel
in 2007 was used as a
quasi-experiment.
CMS, located in the
CCG, was the only
NASCAR racetrack in
North Carolina that held
races every year during
our sample period.
Races occurred bi-
annually, in October
and May, ensuring that
all full and near full-
term births in the
sample were prenatally
exposed via the mother
to at least one
NASCAR event.

were classified as the
treatment group, while the
control group consists of
births where the mother's
residential address is in the
CCG but is at least 10,000m
from the racetrack centroid

BWwas the newborn's
weight, in grams. LBW
was defined as BW
<2500 g. SGA was
defined as BW below
the tenth percentile for
clinical GA.

Age at outcome: birth

a non-randomized
treatment group before
and after deleading to
those in the control group.
Models were adjusted for
mother's age, education,
race, and smoking
behavior; father's age,
education, and race;
infant's birth order and
sex; as well as proximity to
a TRI facility or airport,
median household income,
and age of housing stock;
a set of census tract,
month, and year indicator
variables were also
included

BW (g)

All births

Any exposure: 102. 5
(45.73, 152.2)

Trimester 1: 418.6 (205.1,
632.1)

Trimester 2: 47.68 (-40.01,
135.4)

Trimester 3: 262 (97.01,
427.1)

Full-term births

Any exposure: 24.08
(-15.14, 63.29)

Trimester 1: 104.7 (-54.65,
264)

Trimester 2: 44.16 (-36.35,

124.7)

Trimester 3: 80.19 (-30.44,

190.8)

LBW
All births

Any exposure: -0.045
(-0.07, -0.019)

Trimester 1: -0.062
(-0.178, 0.054)

Trimester 2: -0.022
(-0.061, 0.017)

Trimester 3: -0.158
(-0.314, -0.001)

Full-term births

Any exposure: 0.001
(-0.014, 0.016)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Trimester 1: 0.05 (-0.038,
0.138)

Trimester 2: -0.035
(-0.054, -0.016)

Trimester 3: -0.006 (-0.07,
0.057)

SGA
All births

Any exposure: -0.04
(-0.064, -0.016)

Trimester 1: -0.042
(-0.242, 0.158)

Trimester 2: -0.058
(-0.118. 0.002)

Trimester 3: -0.038
(-0.122, 0.045)

Full-term births

Any exposure: -0.028
(-0.051, -0.004)

Trimester 1: -0.053
(-0.274, 0.168)

Trimester 2: -0.049
(-0.103, 0.004)

Trimester 3: 0.022 (-0.081,
0.125)

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Reference and Study
Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

AAS = atomic absorption spectrometry; AD = abdominal diameter; ALSPAC = Avon Longitudinal Study of Parents and Children; As = arsenic; BL = birth length; BMI = body mass
index; BW = birth weight; BWGA = birth weight-for-gestational age; BWZ = birth weight Z-score; C-ABCS = China-Anhui Birth Cohort Study; CANDLE = Conditions Affecting
Neurocognitive Development and Learning in Early Childhood; CC = chest circumference; CCG = Charlotte-Concord-Gastonia; Cd = cadmium; CD = cephalic diameter;
CHL = crown-heel length; CMS = Charlotte Motor Speedway; Cr = chromium; d = day(s); EMASAR = Study on the Environment and Reproductive Health; e-REACH = e-waste
Recycling Exposure and Community Health; ETS = environmental tobacco smoke; FLEHS = Flemish Environment and Health Study; GA = gestational age; GFAAS = graphite
furnace atomic absorption spectrometry; HC = head circumference; Hg = mercury; HOME = Health Outcomes and Measures of the Environment; hr = hour(s); HR-ICP-MS = high
resolution inductively coupled plasma mass spectrometry; ICP-MS = inductively coupled plasma mass spectrometry; ICP-QQQ = inductively coupled plasma triple quad;

INMA = Instituto de Nanociencia y Materiales de Aragon; IQR = interquartile range; IUGR = intrauterine growth restriction; IVF = in vitro fertilization; LA-ICP-MS = laser ablation-
inductively coupled plasma-mass spectrometry; LBW = low birth weight; LGA = large for gestational age; LIFE = Longitudinal Investigation of Fertility and the Environment;
LMP = last menstrual period or last missed period; In = natural log; LOD = limit of detection; MIREC = Maternal-Infant Research on Environmental Chemicals; min = minute(s);
Mn = manganese; mo = month(s); MSA = Metropolitan Statistical Area; NICE = Nutritional impact on Immunological maturation during Childhood in relation to the Environment;
OR = odds ratio; PI = Ponderal Index; PROGRESS = Programming Research in Obesity, Growth Environment and Social Stress; PROTECT = Puerto Rico Test site for Exploring
Contamination Threats; QL = lower quartile; RR = relative risk; SD = standard deviation; SES = socioeconomic status; SGA = small for gestational age; TRI = Toxics Release
Inventory; UCB = umbilical cord blood; WHEALS = Wayne County Health, Environment, Allergy and Asthma Longitudinal Study; wk = week(s); yr = year(s).
aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect
estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated
interval. Categorical effect estimates are not standardized.
bEffect estimates unable to be standardized.

°Pb measurements were converted from |jg/L to |jg/dL.
dNo cut points provided for the categorizations.

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Table 8-5

Epidemiologic studies of Pb exposure and preterm birth

Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Xu eta!. (2012)

Guiyu and Xiamen
China

2001-2008

Cohort

n: 531 (n = 432 from Guiyu
and n = 99 from Xiamen)

Women who gave birth in
Guiyu or non-urban area of
Xiamen between 2001 and
2008

Cord blood

UCB measured by GFAAS

Age at measurement:
birth

Median:

Guiyu: 10.78 pg/dL
Xiamen: 2.25 [jg/dL
Max:

Guiyu: 47.46 |jg/dL
Xiamen: 7.22 [jg/dL

Preterm birth rate	Multiple logistic regression

models were adjusted for
Preterm birth was defined maternal age and infant sex
as birth <37 wk gestation

Age at outcome: birth

OR (95% Cl)b: 1.09
(0.93, 1.28)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Xu et al. (2022b)
Argentina
2011-2012
Cross-sectional

EMASAR
696

n

Women who either were
about to deliver or had given
birth within the last 48 hr at
one of the two hospitals.
Women had to be above
18 yr of age.

Blood

Maternal blood measured
by ICP-MS

Age at measurement:
birth

Median0:

Overall: 1.34 pg/dL
Ushuaia: 0.98 [jg/dL
Salta 1.50 pg/dL

Geometric mean0:

Overall: 1.393 pg/dL
Ushuaia: 1.01 pg/dL
Salta 1.58 pg/dL

75th°:

Overall: 1.851 pg/dL
Ushuaia: 1.30 pg/dL
Salta: 2.09 pg/dL

Preterm birth

Medical records were
used to obtain measures
at birth.

Age at outcome: birth

Logistic models adjusted for
maternal age, pre-pregnancy
BMI, parity, smoking,
education, and LBW

OR (95%CI)

T1
T2
T3

Reference
1.24 (0.35,
1.26 (0.32,

4.40)
5.00)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Freire et al. (2019)
Spain

2000-2008
Cross-sectional

INMA Project
n: 327

Pregnant women of general
population resident in each
study area [Ribera d'Ebre,
Menorca, Granada,

Valencia, Sabadell, Asturias
and Gipuzkoa] and their
children. Criteria for
inclusion of the mothers
were: (1) to be resident in
one of the study areas, (2)
to be at least 16 yr old, (3) to
have a singleton pregnancy,
(4) to not have followed any
program of assisted
reproduction, (5) to wish to
deliver in the reference
hospital and (6) to have no
communication problems

Other: Placenta

Placenta (including
maternal and fetal sides as
well as central and
peripheral parts) measured
by GFAAS with Zeeman
background correction

Age at measurement:
birth

Median: <6.5 ng/g (LOD)
75th: <6.5 ng/g (LOD)

Preterm delivery

Preterm birth was defined
as live birth before 37 wk
of pregnancy,

Age at outcome:
birth

Logistic regression models
were adjusted for cohort
(random effect), newborn sex,
co-exposure to other metals
(As, Hg, Cd, Mn, Cr), and
maternal education level

OR (95% Cl)b:
(0.04, 4.70)

0.40

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Yu eta!. (2019)

Shanxi Province
China

December 2009-
December 2013

Case-control

n: 528

Women with prenatal
examination at <22
gestational weeks, >18 yr
old, and living in the local
counties for >1 yr

Blood

Maternal blood (serum) was
measured by ICP-MS

Age at measurement:
maternal age at first
trimester (<12 wk gestation)
or second trimester (13-
28 wk)

Mediand

Overall: 0.0482 pg/dL

First trimester:

0.0489 pg/dL

Second trimester:
0.0476 pg/dL
75thd:

Overall: 0.0751 pg/dL
First trimester:

0.0783 pg/dL

Second trimester:
0.0735 pg/dL

Spontaneous preterm
birth

Spontaneous preterm
birth is defined as a live
birth at <37 wk GA without
iatrogenic causes,
including spontaneous
preterm labor with intact gender
membranes and PROM

Age at outcome:
birth

Unconditional logistic
regression models were
adjusted for maternal age, BMI,
education, occupation,
residence, gravidity, parity,
spontaneous abortion history,
folic acid use, drug use,
passive smoking, and child

OR (95% Cl)b

Overall: 1.46 (0.97, 2.18)

First trimester: 1.63
(0.91, 2.91)

Second trimester: 1.27
(0.71, 2.28)

Xu et al. (2022a)

Pingding, Shouyang,
and Taigu Counties

Shanxi Province

China

December 2009-
December 2013

Case-Control

n: 148 (74 cases, 74
controls)

Pregnant women were
recruited if over 18 yr old,
living locally for at least 1 yr,
seeking first prenatal visit at
or before 22 gestational
weeks, and seeking to
manage birth/pregnancy at
Maternal and Child Health
Hospitals of study counties.

Blood

Maternal blood (serum)
measured by ICP-MS

Age at measurement:
maternal age during 4-22
gestational week

Mediand: 0.049 pg/dL
75thd: 0.078 pg/dL

Spontaneous preterm
birth

Information about
spontaneous preterm birth
was collected from
pregnancy health records
at the hospitals

Age at outcome: birth

Unconditional logistic
regression with adjustment for
age, BMI, education,
occupation, residence,
gravidity, parity, spontaneous
abortion history, folic acid use,
medication use, passive
smoking, infant sex, fasting
blood collection, and sampling
time.

OR (95% CI):

Q1
Q2
Q3
Q4

Reference
1.63 (0.53, 5.04)
1.81 (0.60, 5.52)
4.09 (1.31, 12.77)

p for trend: 0.017

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Tsuii eta!. (2018)
Japan

January 2011 and
March 2014

Cohort

JECS
n: 14,847

Women who delivered live
birth infant with singleton
pregnancies without missing
exposure or covariate data

Blood

Maternal blood measured
by ICP-MS

Age at measurement:
Maternal age at gestational
weeks 14-39 (mean
maternal age 31.4 yr)

Preterm birth

Preterm births were
divided into early (<34 wk)
and late preterm births
(34 to <37 wk)

Age at outcome:
birth

Multivariable logistic regression
analysis adjusted forage, pre-
pregnancy BMI, smoking,
smoking habits of partner,
drinking habits, gravidity,
parity, the number of cesarean
sections, uterine infection,
household income, educational
levels, and sex of infant

OR (95% CI)
Early preterm

Q1
Q2
Q3
Q4

Reference
0.66 (0.37, 1.20)
0.80 (0.46, 1.41)
1.22 (0.74, 2.02)

p for trend: 0.134
Late preterm

75th: 7.44 ng/g

Q1: Reference



Q2: 0.99 (0.78, 1.26)

Quartiles (ng/g)

Q3: 0.98 (0.77, 1.25)

Q1: <4.49

Q4: 0.92 (0.72, 1.18)

Q2: 4.80-5.95

p for trend: 0.920

Q3
Q4

5.96-7.43
>7.44

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RefereDCesignnd	Study Population	Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Goto et al. (2021)
Japan

January 2011 to
March 2014

Cohort

JECS
n: 15,540

First, data from participants
who withdrew from the study
or did not meet the Pb
measurement quality control
criteria were excluded
(n = 2,002). Second, data
from mothers who: were lost
to follow-up; had severe
maternal conditions
preceding pregnancy, such
as chronic hypertension,
pregestational diabetes or
cardiac disease, during
pregnancy; or had
pregnancies ending in
abortions or stillbirths
(n = 1,209) was excluded.
Third, data from infants with
chromosomal or major
congenital anomalies
(n = 263) or multiple births
(n = 283) was excluded.

Blood

Maternal blood measured
by ICP-MS

Age at measurement:
Maternal age at second or
third trimester (mean age at
delivery: 31 ± 5.0 yr)

Mean: 0.69 [jg/dL
Median: 0.63 [jg/dL
75th: 0.78 pg/dL
Max: 7.4 pg/dL

Preterm birth (<37
gestational weeks) risk

Preterm birth was defined
as a GA of less than 37
completed wk.

Age at outcome:
birth

Multivariable linear regression
models were adjusted for
maternal age at birth, BMI
before pregnancy, weight gain
during pregnancy, maternal
educational background, a
history of preterm birth, alcohol
consumption during pregnancy,
smoking habit during
pregnancy, and parity

OR (95% CI), per
0.1 pg/dL increase in
maternal blood Pb: 0.90
(0.70, 1.16)

OR (95% CI), per
doubling increment in
maternal blood Pb: 0.978
(0.689, 1.39)

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RefereDCesignnd	Study Population	Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Rabito et al. (2014)

Shelby County,
Tennessee
United States

2008-2011

Cohort

CANDLE study
n: 98

Healthy pregnant woman
between the ages of 16 and
40 yr, carrying a single fetus
with the intent to deliver the
fetus, residence within
Shelby County, Tennessee,
and having the intent to
deliver at one of three area-
based hospitals

Blood and cord blood

Maternal blood, collected at
second and third trimester
and at delivery, and cord
blood, collected at deliver,
were measured by ICP-MS

Age at measurement:
Maternal age at collection
(median: 29.50 yr)

Median:

Second trimester:
0.43 [jg/dL

Third trimester: 0.43 |jg/dL
At delivery: 0.50 [jg/dL
Cord blood: 0.37 [jg/dL

Preterm birth

Preterm birth (<37 wk),
early term birth (37-
39 wk), or full-term birth
(>39 wk) based on GA,
which was determined by
expected due data and
LMP

Age at outcome:
birth

Logistic regression models
were adjusted for marital
status, maternal education
level, and maternal income

Geometric mean (SD):
Second trimester: 0.42
(0.20) [jg/dL

Third trimester: 0.45
(0.28) [jg/dL

At delivery: 0.50
(0.35) [jg/dL
Cord blood: 0.37
(0.32) [jg/dL

Max:

Second trimester:
1.22 [jg/dL

Third trimester: 2.10 |jg/dL
At delivery: 2.47 [jg/dL
Cord blood: 1.80 [jg/dL

OR (95% Cl)b, per 0.1-
unit increase in maternal
blood Pb
Preterm birth

Second trimester: 1.66
(1.23, 2.23)

Third trimester: 1.24
(1.01, 1.52)

Early term birth

Second trimester: 0.87
(0.63, 1.20)

Third trimester: 0.88
(0.69, 1.13)

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RefereDCesignnd	Study Population	Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Taylor et al. (2015)

Bristol
UK

April 1991-December
1992

Cohort

ALSPAC
n: 4,285

All pregnant women in the
former Avon Health
Authority with an expected
delivery date between April
1, 1991, and December 31,
1992, were eligible for the
study

Blood

Maternal blood measured
by ICP-MS, collected as
early as possible in
pregnancy (median GA of
sampling: 11 wk)

Age at measurement:
Maternal age at
measurement

Preterm delivery

Preterm delivery was less
than 37 wk of gestation

Age at outcome:
birth

Logistic regression models for
preterm birth were adjusted for
maternal height, maternal pre-
pregnancy weight, maternal
educational attainment, parity,
number of cigarettes per day,
sex of baby

OR (95% Cl)b:
(1.35, 3.00)

2.00

Mean (SD): 3.67
(1.47) [jg/dL

Geometric mean:
3.43 [jg/dL

Median: 3.42 [jg/dL
Max: 19.14 pg/dL

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Li eta!. (2017a)
China

January 1 to
December 31, 2009

Cohort

C-ABCS
n: 3,125

Mother-and-singleton-
offspring pairs from Hefei
City who provided informed
consent, did not drink
alcohol or smoke cigarettes
during pregnancy, did not
have mental disorders, did
not have pregnancy-induced
hypertension, preeclampsia,
gestational diabetes, heart
disease, thyroid-related
disease, a history of >3
previous miscarriages, or
plans to leave location
before delivery

Blood

Maternal blood (serum)
measured by GFAAS
coupled with a deuterium-
lamp background correction
system, collected in the first
and second trimesters
(median time for serum
collection: 14 gestational
weeks; range from 4 to 27
gestational week)

Mean: 1.50 [jg/dL
Max: 5.46 [jg/dL

Tertiles:

low-Pb: <1.18 |jg/dL

medium-Pb: 1.18-
1.70 [jg/dL

high-Pb: >1.71 pg/dL

Preterm birth

Gestational week was
calculated using mother's
last menstrual period.

Preterm birth was defined
as a live birth at less than
37 completed gestational and parity
weeks and preterm birth
can be further sub-divided
into early preterm birth
(<32 gestational weeks),
moderate preterm birth
(32 to <34 gestational
weeks) and late preterm
birth, 34 to <37
gestational weeks)

Age at outcome:
birth

Multiple logistic regression
models estimated the
association between maternal
serum Pb level and risk of
preterm birth, adjusted for
maternal age, pre-pregnancy

OR (95% CI):

Low-Pb: Reference

Medium-Pb: 2.33 (1.49,
3.65)

High-Pb: 3.09 (2.01,

BMI, monthly income, gravidity, 4.76)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Ashrap et al. (2020)

Puerto Rico

2010-2017

Cohort

PROTECT
n: 731

Participants were recruited
at approximately 14 ± 2 wk
of gestation at seven
prenatal clinics and
hospitals throughout
Northern Puerto Rico and
followed until birth; maternal
age between 18 and 40 yr;
residence inside of the
Northern Karst aquifer
region; disuse of oral
contraceptives within the
3 mo prior to pregnancy;
disuse of IVF to become
pregnant; and free of any
major medical or obstetrical
complications, including pre-
existing diabetes. Each
woman participated in a total
of up to three study visits
(18 ± 2 wk, 22 ± 2 wk, and
26 ± 2 wk of gestation)

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
18-40 (collection between
18 and 26 wk of gestation)

Geometric mean (SD):

Preterm births: 0.39
(1.6) pg/dL

Term births: 0.32
(1.5) pg/dL
Median:

Preterm births: 0.36 pg/dL
Term births: 0.32 pg/dL

Preterm birth (overall and
spontaneous preterm
birth)

All the birth outcome data
were extracted from
medical records. Preterm
birth was defined as <37
completed weeks of
gestation with further
classification of
spontaneous preterm birth
(presentation of
premature rupture of the
membranes, spontaneous
preterm labor, or both)
and non-spontaneous
preterm birth (preterm
births with preeclampsia
or with both artificial
membrane rupture and
induced labor)

Age at outcome:
birth

Logistic regression models
were adjusted for maternal
age, maternal education level,
pre-pregnancy BMI, and
exposure to second-hand
smoking

OR (95% CI)b, per IQR

increase in maternal

blood In-Pb

Preterm birth

Overall: 1.63 (1.17, 2.28)

Spontaneous: 1.53 (1.00,

2.35)

Tertilese:

Overall preterm birth:

T1: Reference

T2: 1.27 (0.65, 2.47)

T3: 1.93 (1.02, 3.62)

Spontaneous preterm
birth:

T1
T2
T3

Reference
0.69 (0.29, 1.66)
1.50 (0.71, 3.18)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Ashrap et al. (2021) PROTECT
n = 682

Puerto Rico

2011-2017
Cohort

Participants were recruited
at approximately 14 ± 2 wk
of gestation at seven
prenatal clinics and
hospitals throughout
Northern Puerto Rico and
followed until birth; maternal
age between 18 and 40 yr;
residence inside of the
Northern Karst aquifer
region; disuse of oral
contraceptives within the
3 mo prior to pregnancy;
disuse of IVF to become
pregnant; and free of any
major medical or obstetrical
complications, including pre-
existing diabetes. Each
woman participated in a total
of up to three study visits
(18 ± 2 wk, 22 ± 2 wk, and
26 ± 2 wk of gestation)

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:
18-40 (collection between
18 and 26 wk of gestation)

Geometric mean: 3.1 [jg/dL
Median: 3.1 [jg/dL
75th 4.1 |jg/dL
95th: 6.5 pg/dL
Max: 15.1 pg/dl_

Preterm birth (overall and Logistic regression models

spontaneous preterm
birth)

Birth outcomes were
extracted from medical
records. Psychosocial
status was evaluated
using four questionnaires

Age at outcome: birth

were adjusted for maternal
age, maternal education, pre-
pregnancy BMI, and exposure
to secondhand smoking

OR (95% Cl)b, per IQR
increase in in maternal
blood In-Pb

Preterm birth:

Good Psychosocial
Status: 1.72 (1.14, 2.58)

Poor Psychosocial
Status: 1.43 (0.69, 2.97)

Spontaneous preterm
birth:

Good Psychosocial
Status: 1.56 (0.93, 2.60)

Poor Psychosocial
Status: 1.22 (0.42, 3.56)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Bui et al. (2022)

North Carolina
United States

2004-2009

Quasi-experimental

CCG MSA

n: 147,673 live births in the
CCG MSA; Treatment group
n: 1,138; Control group n:
13,398

Exogenous variation in Pb
exposure resulting from
NASCAR's deleading of
racing fuel in 2007 was used
as a quasi-experiment.
CMS, located in the CCG,
was the only NASCAR
racetrack in North Carolina
that held races every year
during our sample period.
Races occurred bi-annually,
in October and May,
ensuring that all full and
near full-term births in the
sample were prenatally
exposed via the mother to at
least one NASCAR event.

Births where the mother's
residential address was
within 4,000 meters of CMS
were classified as the
treatment group, while the
control group consists of
births where the mother's
residential address is in the
CCG but is at least
10,000m from the racetrack
centroid

Preterm birth

Preterm birth was defined
as clinical GA <37 wk.

Age at outcome: birth

Difference-in-difference models
were used to compare birth
outcomes in a non-randomized
treatment group before and
after NASCAR deleading to
those in the control group.
Models were adjusted for
mother's age, education, race,
and smoking behavior; father's
age, education, and race;
infant's birth order and sex; as
well as proximity to a TRI
facility or airport, median
household income, and age of
housing stock; a set of census
tract, month, and year indicator
variables were also included

(3 (95% CI)b, as
estimated average
treatment effect of
treatment group
All births

Any exposure: -0.03
(-0.057, -0.002)

Trimester 1: -0.247
(-0.438, -0.057)
Trimester 2: 0.019
(-0.042, 0.079)
Trimester 3: -0.163
(-0.277, -0.049)

ALSPAC = Avon Longitudinal Study of Parents and Children; As = arsenic; BMI = body mass index; C-ABCS = China-Anhui Birth Cohort Study; CANDLE = Conditions Affecting Neurocognitive
Development and Learning in Early Childhood; CCG = Charlotte-Concord-Gastonia; Cd = cadmium; CMS = Charlotte Motor Speedway; Cr = chromium; EMASAR = Study on the Environment
and Reproductive Health; GA = gestational age; GFAAS = graphite furnace atomic absorption spectrometry; hr = hour(s); Hg = mercury; ICP-MS = inductively coupled plasma mass
spectrometry; INMA = Instituto de Nanociencia y Materiales de Aragon; IVF = in vitro fertilization; LBW = low birth weight; LMP = last menstrual period or last missed period; LOD = limit of
detection; mo = month(s); MSA = Metropolitan Statistical Area; OR = odds ratio; PROM = premature rupture of membranes; PROTECT = Puerto Rico Test site for Exploring Contamination
Threats; SD = standard deviation; TRI = Toxics Release Inventory; UCB = umbilical cord blood; wk = week(s); yr = year(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect estimates are

standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated interval. Categorical effect

estimates are not standardized.

bEffects estimates unable to be standardized.

°Pb measurements were converted from |jg/L to |jg/dL.

dPb measurements were converted from ng/mLto |jg/dL.

eNo cut points provided for the categorizations.

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Table 8-6

Epidemiologic studies of Pb exposure and birth defects

Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Jin et al. (2013)

Shanxi Province
China

October 2002 -
onward

Case-control

n: 210: 80 controls, 50 any
NTD case; 36 cases of
anencephaly; and 44 cases
of spina bifida

Once a fetus with an NTD
was identified as a case, a
healthy newborn without
congenital malformations
was selected as a control.
The control was of the same
sex as the case and had a
mother residing in the same
county as that of the case. In
this study, we randomly
selected 36 cases of
newborns with anencephaly
and 44 cases of newborns
with spina bifida as case
groups and 50 healthy term
newborns as a control group.

Other: Placenta

Placental tissue, collected
at delivery or pregnancy
termination, was
measured with ICP-MS

Age at Measurement:
delivery or pregnancy
termination

Mean (SD)

Controls: 22.38 (16.35)
ng/g; NTD cases: 23.30
(22.42) ng/g;

Anencephaly cases:
19.30 (15) ng/g

Spina bifida cases: 23.04
(20.03) ng/g

Median

Controls: 16.9 ng/g

NTD cases: 17.59 ng/g
Anencephaly cases:
10.96 ng/g Spina bifida
cases: 17.38 ng/g
75th:

Controls: 28.83 ng/g
NTD cases: 28.15 ng/g
Anencephaly cases:
28.86 ng/g

Spina bifida cases:
28.86 ng/g

Birth defects: NTDs

Trained local health
workers made primary
diagnoses by physical
examination of the
fetal/newborn body for
any pregnancy
outcomes and filled in
a reporting form for
each case. Three
pediatricians
independently
reviewed the case
report forms and
photographs before
assigning the final
diagnostic codes

Age at outcome:
birth or pregnancy
termination

No attempt was made
to adjust for
confounding factors in
our analyses of Pb
because no differences
in their placental
concentrations were
present between cases
and controls.

OR (95% Cl)b:

Any NTD: 1.14 (0.56, 2.30)

Anencephaly: 1.08 (0.46,
2.56)

Spina bifida: 1.19 (0.53, 2.67)

Liu et al. (2021)

n: 332

Other: Umbilical cord
tissue

Birth defects: NTDs

Multivariate logistic
regression model

OR (95% CI)b:
1.94)

1.23 (0.78,

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Shanxi, China

2004-2016

Case-control

Fetuses from elective
pregnancy terminations and
newborns from the Shanxi
Province in China. Cases
were defined as those with
NTD, and controls were
healthy newborns matched
by maternal residence and
date of last menstruation.

Umbilical cord tissue
measured by ICP-MS

Age at measurement:
At delivery or elective
termination

Median: 26.18 ng/g
75th: 48.58 ng/g
Max: 225.572 ng/g

NTD cases were
diagnosed by fetal
ultrasound scan or
physical examination
at birth or pregnancy
termination.

Age at outcome:
birth or pregnancy
termination

adjusted for folic acid
supplementation

Categorization:

Low exposure
(<1.10 ng/g)

High exposure
(>=1.10 ng/g)

Tian etal. (2021)

Shanxi province,
China

2003-2016

Case control

n: 750

Participants were recruited
from six counties or cities in
the Shanxi province of
northern China.

Blood

Maternal blood (serum)
was measured by ICP-MS

Age at measurement:
Maternal age at collection

Median0:

Controls: 0.087 [jg/dL
Case: 0.115 |jg/dL

75thc:

Controls: 0.197 [jg/dL
Cases: 0.268 [jg/dL

Birth defects: NTDs

Diagnoses of
malformation are
made by local health
workers through
physical examination
of the newborns or
electively terminated
fetuses, in

combination with fetal
ultrasound scans.

Age at outcome: birth
or pregnancy
termination

Multilevel mixed effects
logistic regression
model adjusted for
maternal age, maternal
BMI, education,
gestational weeks, sex
of the fetus,
periconceptional folic
acid use, maternal flu,
or fever.

OR (95% CI):

Tertilesd
NTDs

Lowest: Reference
Medium: 2.05 (1.05, 4.02)
Highest: 3.51 (1.76, 6.98)
p for trend: <0.001

Spina bifida
Lowest: Reference
Medium: 2.16 (1.00, 4.88)

Highest: 5.16 (2.24,
p for trend: 0.022

Anencephaly
Lowest: Reference

11.87)

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Reference and
Study Design

Study Population	Exposure Assessment	Outcome

Confounders	Effect Esti™*fs and 95%

Pi etal. (2018)

Shanxi Province
(Pingding, Xiyang,
Taigu, and Zezhou)
China

2005-2007

Case-control

Medium: 2.97 (1.09, 8.12)
Highest: 5.54 (1.89, 16.19)
p for trend: 0.002

Female Infants
NTDs

Lowest: Reference
Medium: 2.63 (0.99, 7.24)
Highest: 6.45 (2.20, 18.95)
p for trend: 0.001

Male Infants
NTDs

Lowest: Reference

Medium: 2.11 (1.02, 4.34)
Highest: 2.16 (1.03, 4.59)
p for trend: 0.048

n: 103 cases and 206
controls

Newborns or terminated
fetuses with any major
external structural defects,
including OFCs, NTDs,
congenital hydrocephalus,
limb defects were recruited
from five rural counties in
Shanxi Province

Other: Placenta

Placental tissue, collected
immediately after delivery,
was measured by ICP-MS

Age at Measurement:
birth

Mean (SD)

Controls 72.6 (34.8) ng/g
Case: 130.9 (95.7) ng/g
Median

Controls: 67.9 ng/g
Cases: 96.1 ng/g
75th

Controls: 98.1 ng/g
Cases: 176.4 ng/g

Birth defects: OFCs

Diagnoses of
newborns/fetuses with
major birth defects
were done through
physical examination
or prenatal ultrasound
examination by county
healthcare workers.
Once a newborn/fetus
with a major birth
defect was identified
as a case, a healthy
newborn with no
congenital
malformation was
selected as a control
to match the case by
residence of the
mother (the same

Binary logistic
regression adjusted for
occupation, newborn
sex, gestational weeks,
previous history of birth
defects, maternal flu or
fever, and passive
smoking during the
periconceptional period

OR (95% CI)
Orofacial defects:
T1: Reference
T2: 3.88 (1.78, 8.42)
T3: 5.17 (2.37, 11.29)
p for trend: <0.001

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa





Tertiles (ng/g):
T1: <57.5
T2: 57.5-96.8
T3: >96.8

county), date of the
LMP (±4 wk), and
newborn sex.

Age at outcome:
at diagnosis





Takeuchi et al.

Japan

2011-2014

Case-control

JECS

n:192 cases, 1920 matched
controls

Pregnant women living in the
study area and
understanding of the
Japanese language.
Participants were excluded if
they had missing data
(heavy metal data, matching
variables, and/or both).
Covariates for matching
were maternal age,
psychological stress
measured by the K6 score,
gestational weeks of blood
sampling during second
trimester, folic acid intake
estimated from a food-
frequency questionnaire,
alcohol intake (self-reported),
smoking (self-reported),
education level, BMI before
pregnancy, diabetes before
pregnancy, intake of
supplements (self-reported),
and regional center

Blood

Maternal blood measured
by ICP-MS

Age at measurement:

Maternal age at collection
(second trimester)

Mediane

Cohort: 0.585 [jg/dL
Cases: 0.584 [jg/dL
Controls: 0.575 [jg/dL
75the

Cohort: 0.73 [jg/dL
Cases: 0.72 [jg/dL
Controls: 0.71 [jg/dL

Birth defects: Cleft
palate and cleft lip
(isolated)

Validated medical
records were used to
identify isolated cleft
lip and palate.

Conditional logistic
regression adjusted for
sex and concentrations
of Hg, Cd and Mn

OR (95% CI), per 0.1 pg/dL
increase in maternal blood
Pb: 1.10 (0.55, 2.21)

Mivashita et al.

(2021)

JECS.
N: 89,273

Blood

Birth defects:
Abdominal congenital
malformations

Multivariate logistic OR (95% CI)
regression models were Abdominal congenital
adjusted for maternal malformations
age, smoking habit,

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Japan

January 2011-
2014

Cohort

¦March

Pregnant women and their
newborns recruited for the
JECS. Singleton, live births
were included.

Maternal blood (serum)
measured by ICP-MS.

Age at measurement:
maternal age at collection
(mid-late pregnancy)

Median

Cohort: 5.84 ng/g
Controls: 5.85 ng/g
Cases: 5.53
75th

Cohort: 7.32 ng/g
Controls: 7.32 ng/g
Cases: 7.00 ng/g
Max

Cohort: 110 ng/g
Quartiles (ng/g):

Q1
Q2
Q3
Q4

<4.7

4.7-<5.84

5.84-<7.32

>7.32

Abdominal congenital
malformations
(including
omphalocele,
gastroschisis,
esophageal atresia
with/without fistula,
duodenal atresia,
intestinal atresia,
anorectal atresia,
diaphragmic hernia)
were identified from
birth records or
records 1 mo post
birth

Age at outcome: birth
to month post birth

drinking habit, paternal
smoking habit, birth
year of child, sex of
child

Q1
Q2
Q3
Q4

Reference
1.19 (0.76, 1.84)
0.77 (0.47, 1.26)
0.85 (0.52, 1.38)

p for trend: 0.233

Diaphragmic hernia

Q1
Q2
Q3
Q4

Reference
1.24 (0.51, 2.99)
0.89 (0.34, 2.31)
0.81 (0.30, 2.20)

p for trend: 0.543

Omphalocele

Q1
Q2
Q3
Q4

Reference
0.72 (0.29, 1.81)
0.35 (0.11, 1.12)
0.35 (0.11, 1.13)

p for trend: 0.033

Gastroschisis
Q1: Reference

Q2
Q3
Q4

1

1.00 (0.14, 7.09)
2.63 (0.50, 13.70)

p for trend: 0.212

Esophageal atresia
with/without fistula

Q1
Q2
Q3
Q4

Reference
0.49 (0.04, 5.43)
0.95 (0.13, 6.80)
1.88 (0.33, 10.50)

p for trend: 0.346

Duodenal atresia/stenosis

Q1
Q2
Q3

Reference
0.25 (0.03, 2.27)
0.50 (0.09, 2.75)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Q4: 0.99 (0.24, 4.06)
p for trend: 0.910

Intestinal atresia/stenosis

Q1: Reference
Q2: 1.40 (0.31, 6.29)
Q3: 1.06 (0.21, 5.27)
Q4: 1.12 (0.22, 5.64)
p for trend: 0.989

Anorectal atresia/stenosis
Q1: Reference
Q2: 1.65 (0.74, 3.67)
Q3: 0.57 (0.19, 1.68)
Q4: 0.62 (0.21, 1.83)
p for trend: 0.158

Liu et al. (2018)
China

February 2010-
October2011

Case-control

n: 97 cases with CHDs and
201 controls without any
abnormalities

Eligible fetuses with cardiac
defects diagnosed during
prenatal examination were
recruited as the case group.
For each case, one pregnant
control without any fetal
malformation was selected in
the same hospital with a
gestation age within 2 wk of
the case fetus. Cases and
controls with GAs from 14 to
40 wk were selected for this
study after the following
exclusion criteria were
applied: (1) multiple
pregnancies; (2) CHD family
history; (3) fetus diagnosed
with a chromosomal
abnormality or hereditary
syndrome; (4) fetus with

Cord blood

UCB (serum) was
measured by ICP-MS

Age at Measurement:
birth

Median0

Cases: 0.791 [jg/dL

Controls: 0.740 ug/dL
75thc

Case: 0.922 [jg/dL
Controls: 0.877 [jg/dL

Tertiles (|jg/dL):
Low: <0.696
Medium: 0.696-0.826
High: >0.826

Birth defects: CHDs

Cardiac defects
diagnosed during
prenatal examination
were recruited as the
case group.

Age at outcome: age
at diagnosis

Logistic regression
models were adjusted
for maternal age,
maternal pre-pregnancy
BMI, maternal
education level, folic
acid supplement, and
parental smoking

OR (95% CI)
CHD, Overall
Low: Reference
Medium: 1.46 (0.77, 2.77)
High: 1.67 (0.88, 3.17)

Septal Defects
Low: Reference
Medium: 1.20 (0.57, 2.52)
High: 1.61 (0.78, 3.32)

Conotruncal Defects
Low: Reference
Medium: 1.35 (0.60, 3.06)
High: 1.47 (0.65, 3.34)

Right-sided Outflow Tract
Deformity

Low: Reference

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Reference and
Study Design

Study Population	Exposure Assessment	Outcome

Confounders	Effect Esti™*fs and 95%

extra cardiac malformations;
(5) uncompleted
questionnaire for some
reason. CHD cases were
classified into six subtypes
based on the anatomic
lesion: (i) septal defects, (ii)
conotruncal defects, (iii) left
sided outflow tract deformity,
(iv) right-sided outflow tract
deformity, (v) anomalous
pulmonary venous return,
and (vi) other cardiac
structural abnormalities.

Medium: 0.92 (0.37, 2.26)
High: 1.21 (0.50, 2.94)

Left-sided Outflow Tract
Deformity
Low: Reference
Medium: 2.29 (0.62, 8.41)
High: 1.32 (0.29, 5.91)

Anomalous Pulmonary
Venous Return
Low: Reference
Medium: 1.71 (0.37, 7.83)
High: 1.49 (0.30, 7.44)

Other Cardiac Structural
Abnormalities

Low: Reference

Medium: 1.10 (0.36, 3.40)

High: 1.41 (0.47, 4.22)

BMI = body mass index; Cd = cadmium; CHD = congenital heart diseases/defects; GA = gestational age; Hg = mercury; ICP-MS = inductively coupled plasma mass spectrometry;
K6 = Kessler Psychological Distress Scale; LMP = last menstrual period or last missed period; Mn = manganese; mo = month(s); NTD = neural tube defect; OFC = orofacial cleft;
OR = odds ratio; SD = standard deviation; UCB = umbilical cord blood; wk = week(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect

estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated

interval. Categorical effect estimates are not standardized.

bEffects estimates unable to be standardized.

°Pb measurements were converted from ng/mL to |jg/dL.

dNo cut points provided for the categorizations.

ePb measurements were converted from |jg/L to |jg/dL.

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Table 8-7 Epidemiologic studies of Pb exposure and fetal and infant mortality and spontaneous abortion and
pregnancy loss

Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates
and 95% Clsa

Xu eta!. (2012)

Guiyu and Xiamen
China

2001-2008

Cross-sectional

n: 531 (n = 432 from Guiyu
and n = 99 from Xiamen)

Women who gave birth in
Guiyu or non-urban area of
Xiamen between 2001 and
2008

Cord blood

UCB measured by
GFAAS

Age at Measurement:
birth

Median:

Guiyu: 10.78 pg/dL

Xiamen: 2.25 [jg/dL
Max:

Guiyu: 47.46 |jg/dL
Xiamen: 7.22 [jg/dL

Stillbirth rate

Stillbirth was defined as
fetal death before complete
expulsion or extraction from
the mother at >20 wk of
gestation

Age at outcome:
birth

Multiple logistic regression
models were adjusted for
maternal age and infant sex

OR (95% CI)b: 4.20
(3.40, 5.18)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates
and 95% Clsa

Louis et al. (2017)

Michigan and Texas
United States

2005-2009

Cohort

LIFE Study
n: 344

Female partners aged 18-40
and male partners aged
>18 yr who were in a
committed relationship; no
physician diagnosis of
infertility/sterility; off
contraception <2 mo; and an
ability to communicate in
English or Spanish. Female
partners also had to have
menstrual cycles ranging
between 21 and 42 d as
required by the fertility monitor
and without the use of
injectable hormonal
contraceptives in the past year
given the uncertain timing for
ovulation return

Blood

Blood from female and
male partners was
measured by ICP-MS

Age at Measurement:
18-40 for females and
>18 for males

Median

Females: 0.66 [jg/dL
Males: 1.00 [jg/dL

75th:

Females: 0.82 [jg/dL
Males: 1.37 [jg/dL

Pregnancy loss

Pregnancy was
prospectively captured by
women's use of the
Clearblue® digital home
pregnancy test, which is
sensitive in detecting
25 mlU/mL ofhCGand
accurately used by women.
Depending upon timing of
loss, it was detected by
conversion to a negative
pregnancy test, clinical
confirmation, or return of
menses.

Age at outcome:

18-40 yr

Cox proportional hazard
models; individual partner
model adjusted for age, BMI,
history of prior loss conditional
on gravidity, average number
of daily alcoholic drinks
consumed, and cigarettes
smoked during the
preconception and early
pregnancy windows for
females and preconception for
males; couples based model
adjusted for each partner's
metal concentration, age,
difference in couples' ages,
BMI, average number of daily
alcoholic drinks consumed and
cigarettes smoked during the
preconception and early
pregnancy window for females
and preconception for males,
and history of prior loss
conditional on gravidity

HR (95% Cl)b
Individual partner
model

Female partner:
1.01 (0.82, 1.25)
Male partner: 0.95
(0.77, 1.17)
Couple based
model

Female partner:
1.01 (0.80, 1.28)

Male partner: 0.96
(0.77, 1.22)

Viqeh et al.

Tehran
Iran

March 2016-October
2017

Cohort

Tehran Environment and
Neurodevelopmental Disorder

n: 166 (spontaneous abortion
n: 25 and ongoing pregnancy
n: 141)

Pregnant women with GA of
10-16 wk and of Iranian
nationality and Tehran city
inhabitant were invited to
participate in the study.

Blood

Maternal blood was
measured using ICP-MS

Age at measurement:
maternal age at first
trimester

Mean0: 4.96 |jg/dL
Maxc: 70.982 [jg/dL

Spontaneous abortion

Spontaneous abortion
defined as fetal demise
before 20 wk gestation and
reported by study
participant or research
hospital.

Age at outcome: before
20 wk of gestation

Logistic regression models
adjusted for maternal age,
primipara, and previous
abortion

OR (95% CI), per
0.1 [jg/dL increase
in maternal blood
Pb: 1.08 (0.98, 1.20)

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Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates
and 95% Clsa

Tolunay et al.

Ankara
Turkey

January 2012 and July
2012

Cohort

n: 101

The study group consisted of
patients with ongoing
pregnancy (n = 20) and the
reference group consisted of
patients experienced ART
failure, miscarriage, or
biochemical pregnancy
(n = 81)

Blood

Maternal blood was
measured by AAS

Age at Measurement:
20-40

Median

Study group: 2.34 [jg/dL

Reference: group
5.11 [jg/dL

Max

Study group: 7.97 [jg/dL

Reference group:
10.47 [jg/dL for
reference group

Pregnancy loss

Clinical pregnancy was
defined as the presence of
an embryo with a heartbeat
at 6th gestational week.
Ongoing pregnancy was
defined when the
pregnancy had completed
20 wk of gestation.
Implantation rate was
calculated separately for
each woman as the number
of gestational sacs divided
by the number of
transferred embryos
multiplied by 100.

Age at outcome:
completion of 20 wk of
gestation

Log binominal regression
analysis adjusted for age and
BMI

RR (95% CI): 0.978
(0.957, 0.999)

Li et al. (2022)

Hefei
China

October 2019 -
January 2020

Cohort

n: 1184

Participants were selected
from First Affiliated Hospital of
Anhui Medical University while
seeking IVF treatment and
diagnosed with infertility with
their partner. Inclusion criteria:
women were aged between
20 and 45 yr; couples were
diagnosed with infertility
(failure to establish a clinical
pregnancy with unprotected
intercourse for at least 1 yr);
and IVF indicators were tubal
factor, ovulation failure, or
other factors for female

Blood

Maternal blood (serum)
was measured by ICP-
MS

Age at measurement:

maternal age at
collection (day before
oocytes were retrieved
for IVF); mean age was
30.22 yr

Geometric meand:
0.0877 [jg/dL

Mediand: 0.0924 [jg/dL

Spontaneous abortion

Spontaneous abortion
before gestational week 12
was followed upon the 65th
day after embryo transfer.

Age at outcome: maternal
age at outcome (before
gestational week 12)

Logistic regression model for
successful implantation
adjusted for: maternal age,
BMI, treatment protocol,
numbers of retrieved oocytes,
embryo quality

OR (95%CI)b:
Spontaneous
abortion: 1.39 (1.02,
1.91)

Tertiles

Low: Reference

Medium: 1.49 (0.84,
2.63)

High: 1.55 (0.87,
2.79)

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Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates
and 95% Clsa

partner or male factor or
unexplained fertility.

75thd: 0.14399 pg/dL

Tertilesd (pg/dL):
Low: 0.002-0.065
Medium: 0.065-0.125
High: 0.125-0.481

AAS = atomic absorption spectrometry; BMI = body mass index; d = day(s); GFAAS = graphite furnace atomic absorption spectrometry; hCG = human chorionic gonadotropin;
HR = hazard ratio; ICP-MS = inductively coupled plasma mass spectrometry; IVF = in vitro fertilization; mo = month(s); OR = odds ratio; UCB = umbilical cord blood; wk = week(s);
yr = year(s).

aEffect estimates are standardized to a 1 pg/dL increase in blood Pb or a 10 pg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect estimates are

standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated interval. Categorical

effect estimates are not standardized.

bEffects estimates unable to be standardized.

°Pb measurements were converted from pg/L to pg/dL.

dPb measurements were converted from ng/Lto pg/dL.

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Table 8-8

Epidemiologic studies of Pb exposure and placental function

Reference and Study
Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Al-Saleh et al.

Al-Kharj
Saudi Arabia

2005-2006

Cross-sectional

n: 1,578

Women aged 16-50 yr
who delivered in Al-
Kharj hospital, Saudi
Arabia

Blood, cord blood, and
other: placenta

Maternal blood, UCB, and
placental tissue
measured by AAS

Age at Measurement:
maternal age 16-50; birth

Mean ± SD:

Maternal blood:
2.897 ± 1.851 [jg/dL
UBC:

2.551 ±2.592 pg/dL
Placenta:

0.579 ±2.176 pg/g
Median:

Maternal blood:
2.540 pg/dL
UCB: 2.057 pg/dL
Placenta: 0.450 pg/g
75th:

Maternal blood:
3.314 pg/dL

UCB: 2.689 pg/dL

Placenta: 0.630 pg/g
Max:

Maternal blood:
25.955 pg/dL

UCB: 56.511 pg/dL

Placenta: 78 pg/g

Placental function:
Placental thickness

Placental weight and
placental thickness were
recorded by obstetrician in
delivery room

Age at outcome:
birth

Logistic regression model was
adjusted for maternal age, parity,
mother's third trimester BMI,
urinary cotinine, mother's highest
education, total family income,
and GA

OR (95% Cl)b, per unit
increase in maternal blood
Pb: 1.64 (1.12, 2.41)

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Reference and Study
Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Tsuii

Japan

January 2011-March
2014

Cross-sectional

JECS
n: 16,019

Mothers who delivered
a singleton pregnancy

Blood

Maternal blood, collected
during the second
trimester, was measured
by ICP-MS

Age at Measurement:
maternal age at second
trimester

Median: 5.96 ng/g
75th: 7.45 ng/g

Quartiles:

Q1: <4.79 ng/g
Q2: 4.80-5.95 ng/g
Q3: 5.96-7.44 ng/g
Q4: >7.45 ng/g

Placental function:
Placenta previa and
placenta accreta

Data for those with and
without placenta previa
and placenta accreta were
obtained from medical
records.

Age at outcome:
maternal age at diagnosis

Multivariable logistic regression
models were adjusted for age,
smoking, smoking habits of the
partner, drinking habits, gravidity,
parity, number of cesarean
deliveries, and geographic region;
Placenta previa was added as a
covariate when comparisons
were performed with or without
placenta accreta

OR (95% CI):
Placenta previa

Q1
Q2
Q3
Q4

Reference
2.59 (1.40, 4.80)
1.32 (0.66, 2.64)
1.34 (0.67, 2.67)

p for trend: 0.007

Placenta accreta

Q1
Q2
Q3
Q4

Reference
1.46 (0.57, 3.76)
1.68 (0.66, 4.24)
0.79 (0.27, 2.30)

p for trend: 0.345

AAS = atomic absorption spectrometry; BMI = body mass index; CI = confidence interval; GA = gestational age; ICP-MS = inductively coupled plasma mass spectrometry; JECS = Japan
Environment and Children's Study; OR = odds ratio; Q = quartile; SD = standard deviation; UCB = umbilical cord blood; yr = year(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect estimates are
standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated interval. Categorical effect
estimates are not standardized.
bEffect estimates unable to be standardized.

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Table 8-9

Epidemiologic studies of Pb exposure and other pregnancy and other birth outcomes

Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Ashley-Martin et al.

Vancouver, Edmonton,
Winnipeg, Sudbury,
Ottawa, Kingston,
Toronto, Hamilton,
Montreal, and Halifax
Canada

2008-2011

Cohort

MIREC study
n: 1,260

Women were recruited from
10 Canadian sites during
their first trimester and
consented to provide urine
and blood samples. Women
were eligible for inclusion if
they were <14 wk gestation
at the time of recruitment,
>18 yr of age, able to
communicate in French or
English, and planning to
deliver at a local hospital

Blood

Maternal blood was
measured by ICP-MS

Age at Measurement:
Maternal age during 1st
and 3rd trimester

Geometric mean (SD):
0.88 (1.61) pg/dL

Quartiles (pg/dL):
Q1: <0.63
Q2: 0.64 to <0.87
Q3: 0.88 to <1.20
Q4: >1.20

Other Pregnancy and Birth
Outcomes: Fetal metabolic
function

Leptin and adiponectin were
measured in plasma from
1363 stored UCB samples by
ELISA using kits from Meso
Scale Discovery. All samples
were above the LOD.

Age at outcome:
birth

Logistic regression models
were adjusted for maternal
age at delivery, pre-
pregnancy BMI, parity, and
BWZ

OR (95% CI)

Low leptin and maternal
blood Pb:

Q1
Q2
Q3
Q4

Reference
0.9 (0.5, 1.6)
0.6 (0.3, 1.1)
0.9 (0.5, 1.5)

High leptin and maternal
blood Pb:

Q1
Q2
Q3
Q4

Reference
1.2 (0.7, 2.1)
1.0 (0.6, 1.8)
1.7 (1.0, 2.9)

Low adiponectin and
maternal blood Pb:

Q1
Q2
Q3
Q4

Reference
1.3 (0.8, 2.2)
0.8 (0.5, 1.4)
1.1 (0.6, 1.9)

High adiponectin and
maternal blood Pb:

Q1
Q2
Q3
Q4

Reference:
0.9 (0.5, 1.5)
1.1 (0.7, 1.9)
0.9 (0.5, 1.5)

Herlin et al, (2019)

n: 194 enrolled of the 221

Blood, cord blood, and

Other Pregnancy and Birth

Multivariable-adjusted

(3 (95% CI)c:



pregnant women

other: placenta

Outcomes: rTL

linear regression models;

UCB: -0.038 (-0.074,

Salta Province (Andean







models with maternal blood

-0.002)



All pregnant women living in



The rTL was measured in

Pb were adjusted for

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Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

part)

Argentina

October 2012-
December 2013

Cohort

the Andean part of the Salta
province northern Argentina
with estimated delivery date
between October 2012 and
December 2013, were
invited to participate

Maternal blood, UCB,
and placenta were
measured using ICP-MS

Age at Measurement:
birth

Median:

Maternal bloodb:
2.1 [jg/dL

UCBb: 1.4 Mg/dL

Placenta: 5.8 [jg/kg

Max:

Maternal bloodb:
9.9 [jg/dL
UCBb: 6.0 [jg/dL
Placenta: 38 [jg/kg

maternal blood leukocytes
(blood samples collected in
late pregnancy, mainly third
trimester), cord blood
leukocytes, and placental
tissue. We obtained high-
quality DNA and measured
rTL in 169 blood samples of
the pregnant women, 99 of
their placentas, and 98 cord
blood samples of their
babies. The rTL was
measured as the ratio
between the signal intensity
of the telomere sequences
and the signal intensity of a
single-copy gene
(hemoglobin (3 chain), using
real-time polymerase chain
reaction.

Age at outcome:
birth

maternal age, pre-
pregnancy BMI, and
education; models with
placenta were also
adjusted for GA at birth;
models with UCB were
adjusted for maternal age,
pre-pregnancy BMI, GA at
birth, and BW.

Maternal blood: 0.026
(-0.043, 0.095)

Placenta: -0.029 (-0.074,
0.016)

Liao et al. (2015)
Taiwan

n: 113

Pregnant women were
recruited from a single
March-December 2010 institution in northern
Taiwan

Cross-sectional

Blood

Maternal blood
(plasma), collected at
the first trimester
(between 10 and 14wk
of gestation), was
measured by ICP-MS

Age at Measurement:
Maternal age at first
trimester (mean age
30.92 ±3.09 yr)

Geometric mean:
0.048 |jg/L

Other Pregnancy and Birth
Outcomes: Fetal nuchal
translucency thickness

Fetal nuchal translucency
thickness was measured at
10-14 wk of gestation by a
gynecologist and three
trained sonographers

Age at outcome:

Age at scan (between
gestational week 10 and 14)

Multiple linear regression
models were adjusted for
maternal age, gestational
weeks, pre-pregnancy BMI,
supplement use, and
medication

(3 (95% CI)c: 0.022 mm
(-0.06, 0.10)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Ashley-Martin et al.

Canada

2008-2011

Cohort

MIREC Study
n: 1256

Pregnant women in Canada
who had singleton, term
birth (>37 wk)

Blood

Maternal blood was
measured by ICP-MS

Age at measurement:

maternal age at first and
third trimester

Median: 0.62 [jg/dL
75th: 1.03 pg/dL
Max: 4.14 pg/dL

Other Pregnancy and Birth
Outcomes: Fetal immune
system biomarkers

Immune system biomarkers
were measured in the
plasma of UCB samples;
TSLP concentrations were
determined using a
commercial antibody kit; IL-
33 concentrations were
analyzed using antibodies
from an R & D systems duo
set; IgE was determined from
ELISA kits

Logistic regression
adjusted for maternal age

OR (95% CI)

Maternal log-io-Pb blood
concentrations with elevated
(>80%) cord blood
concentrations of IL-33 and
TSLP: 0.79 (0.62, 1.01)

Maternal log-io-Pb blood
concentrations with elevated
(>0.5 kU/L) cord blood
concentrations of IgE: 0.99
(0.77, 1.26)

Age at outcome:
birth

Taylor et al. (2014)

Bristol
UK

April 1991-December
1992

Cohort

ALSPAC study
n: 4,285

Blood

Maternal blood was

Pregnant women enrolled in	,,

the ALSPAC study at a measured by ICP-MS

median GA of 11 wk

Age at Measurement:
Maternal age at
measurement (median
GA of sampling: 11 wk)

Median:
Quintile 1
Quintile 2
Quintile 3
Quintile 4
Quintile 5
Max:

2.11 pg/dL
2.82 pg/dL
3.43 pg/dL
4.13 pg/dL
5.00 pg/dL

Other Pregnancy and Birth
Outcomes: Secondary sex
ratio

The sex of the infant was
recorded at birth

Age at outcome:
birth

Logistic regression models
adjusted for maternal and
paternal age, and parity

OR (95% CI)

Q1
Q2
Q3
Q4
Q5

Reference

1.04 (0.86,	1.42)

0.90 (0.70,	1.15)

1.01 (0.79,	1.30)

1.06 (0.82,	1.37)

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Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

Q1: 2.53 [jg/dL
Q2: 3.11 [jg/dL
Q3: 3.71 [jg/dL
Q4: 4.63 pg/dL
Q5: 19.14 [jg/dL

iloom et al.

Michigan (4 counties)
and Texas (12 counties)
United States

2005-2009

Cohort

LIFE
n: 235

Potential participants were
identified, using fishing
license registries or a
commercially available
direct marketing data base,
from 12 counties in Texas
and four in Michigan,
respectively, with presumed
exposure to persistent
organic pollutants. Inclusion
criteria comprised a
committed heterosexual
relationship, women aged
18-40 yr (men >18), English
or Spanish speaker, no use
of an injectable
contraceptive within 12 mo,
and a menstrual cycle
length of 21^2 d.

Blood

Maternal and paternal
blood, collected before
pregnancy (baseline),
were measured by ICP-
MS

Age at Measurement:
>18, maternal mean
age: 29.75 (SD: 3.73) yr
and paternal mean age:
31.52 (SD:4.57) yr

Mean (SD):

Maternal: 0.71
(0.30) [jg/dL
Paternal: 1.13
(0.63) [jg/dL

Median:

Maternal: 0.66 pg/dL
Paternal: 0.98 pg/dL
Max:

Maternal: 2.23 pg/dL
Paternal: 6.43 pg/dL

Other Pregnancy and Birth
Outcomes: Secondary sex
ratio

Women were followed until
delivery when they
completed and returned birth
announcements that
captured date and sex of
birth, weight and length, and
HC. Secondary sex ratio is
the ratio of live male to
female births, reflecting a
male excess.

Age at outcome:
birth

Log-binomial models for
secondary sex ratio: effect
of maternal exposure
adjusted for paternal
exposure, maternal age,
difference in maternal and
paternal age, and maternal
and paternal smoking,
income, race, serum lipids
(mg/dL), and creatinine for
urine (mg/dL); effect of
paternal exposure adjusted
for maternal exposure,
paternal age, difference in
maternal and paternal age,
and maternal and paternal
smoking, income, race,
serum lipids (mg/dL), and
creatinine for urine (mg/dL)

RR (95% CI)
Maternal Exposure:
T1: Reference
T2: 0.97 (0.78, 1.22)
T3: 1.00 (0.81, 1.24)
p for trend: 0.884
Paternal Exposure:

T1
T2
T3

Reference
1.12 (0.89, 1.41)
1.06 (0.84, 1.34)

p for trend: 0.854

Tertiles (pg/dL):
Maternal Blood Pb
T1: <0.55 (<33rd
percentile)

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Reference and Study
Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

T2: 0.55-0.73 (33rd to
67th percentile)

T3: >0.73 (>67th
percentile)

Paternal Blood Pb

T1: <0.84 (<33rd
percentile)

T2: 0.84-1.16 (33rd to
67th percentile)

T3: >1.16 (>67th
percentile)

Tatsuta et al. (2022b)
Japan

January 2011-March
2014 (followed through
birth)

Cohort

JECS
n: 85,171

Pregnant women and their
paternal partners were
recruited from 15 regions of
Japan. Participants
delivered a live infant with
singleton pregnancy and
had child sex information.
Participants were excluded
is they had a stillbirth,
abortion, multiple births, or
withdrew before birth;
missing blood sample
information; missing
confounders; or without
partner's consent and with
paternal age or occupational
exposure to Pb deficits

Blood

Maternal blood was
measured by ICP-MS.

Age at measurement:

maternal age at
collection (middle or late
pregnancy)

Median: 5.85 ng/g
Max: 110 ng/g

Quartiles (ng/g)

Q1
Q2
Q3
Q4

1.20-4.46
4.47-5.39
5.40-6.35
6.36-7.76

Other Pregnancy and Birth
Outcomes: Secondary sex
ratio

Sex of the infant obtained
from the medical record
transcripts by physicians,
midwives, nurses, or trained
research coordinators.

Logistic regression models
were adjusted for maternal
age at parturition, season
of birth, pre-pregnancy
BMI, annual household
income, gravidity, fertility
treatments, score of the
K6, maternal smoking
status during pregnancy,
passive smoking status
during pregnancy, birth
year and study area
(regional center)

OR (95% CI)

Q1
Q2
Q3
Q4
Q5

Reference
1.082 (1.037, 1.129)
1.122 (1.074, 1.171)
1.214 (1.163, 1.268)
1.279 (1.224, 1.336)

Q5: 7.77-110

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Reference and Study
Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and
95% Clsa

AAS = atomic absorption spectrometry; ALSPAC = Avon Longitudinal Study of Parents and Children; BMI = body mass index; BW = birth weight; BWZ = birth weight Z-score; d = day(s);
ELISA = enzyme-linked immunosorbent assay; HC = head circumference; ICP-MS = inductively coupled plasma mass spectrometry; IgE = immunoglobulin E; IL-33 = interleukin-33;

JECS = Japan Environment and Children's Study; K6 = Kessler Psychological Distress Scale; LIFE = Longitudinal Investigation of Fertility and the Environment; LOD = limit of detection;
MIREC = Maternal-Infant Research on Environmental Chemicals; mo = month(s); OR = odds ratio; Q = quartile; RR = relative risk; rTL = relative telomere length; SD = standard deviation;
T# = fertile #; TSLP = thymic stromal lymphopoietin; UCB = umbilical cord blood; wk = week(s); yr = year(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect estimates are

standardized to the specified unit increase for the 10,h-90,h percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated interval. Categorical effect

estimates are not standardized.

bPb measurements were converted from |jg/L to |jg/dL.

°Effect estimates unable to be standardized.

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Table 8-10

Epidemiologic studies of Pb exposure and postnatal growth

Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Cisa

Siqnes-Pastor et al

(2021)

United States

2013-2016

Cross-sectional

NHANES
n: 1,634

Children aged 6-11 yr
old participating in the
2013-2014 and 2015-
2016 NHANES cycles

Blood

Blood was measured by
ICP-MS

Age at measurement: 6-
11 yrold

Median:

Overall: 0.5 [jg/dL
Girls: 0.5 [jg/dL
Boys: 0.5 [jg/dL

75th:

Overall: 0.8 [jg/dL
Girls: 0.7 [jg/dL
Boys: 0.8 [jg/dL
Max:

Overall: 5.8 [jg/dL
Girls: 5.8 [jg/dL
Boys: 5.0 [jg/dL

Postnatal growth: weight,
WC, upper arm length,
standing height, and BMI

Physical examination was
performed to obtain body
measurements.

Age at outcome: 6-11 yr
old

Linear regression
models were adjusted
for total calorie intake,
race, PIR, children's
age, smoker(s) in the
household, outside-of-
school and at-school
activity scores,
children's sex, and co-
exposure to fluoride,
Mn, Hg, and Se

(3 (95% CI)

BMI (kg/m2): -2.092 (-3.227,
-0.957)

Standing height (cm): -3.116
(-5.03, -1.202)

WC (cm): -5.742 (-8.769,
-2.715)

Upper arm length (cm): -1.068
(-1.625, -0.512)

Girls

BMI (kg/m2): -3.204 (-5.654,
-0.754)

Standing height (cm): -2.89
(-6.691, 0.911)

WC (cm): -6.659 (-12.911,
-0.408)

Upper arm length (cm): -1.696
(-2.859, -0.534)

Boys

BMI (kg/m2): -1.959 (-3.45,
-0.467)

Standing height (cm): -3.828
(-6.588, -1.068)

WC (cm): -6.81 (-10.995,
-2.626)

Upper arm length (cm): -0.89
(-1.691, -0.089)

Kuanq et al.

Nanjing
China

n: 395

Students aged 7-11 yr
(grades 2 to 4) were

Blood

Blood was measured by
ICP-MS

Postnatal growth: height,
weight, bust, waistline,
and BMI

General linearized
models were adjusted
for age and gender

(3 (95% CI)

Height (cm): -3.21 (-4.24,
-2.17)

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Cisa

2012

Cross-sectional

recruited from public
primary schools in
Nanjing, an industry city
from East China.
Students with congenital
mental retardation (third-
degree relatives
included) and other
serious diseases were
excluded. Students and
their parents were
informed of the research
content and purpose.
Only completely
matched groups of
samples, including
questionnaire
information, blood
samples, growth indexes
and school
performances, were
included in the study.

Age at Measurement:
7-11 yr

Mean (SD)b: 3.04
(1.72) pg/dL

Medianb: 2.61 pg/dL

Growth: Individual
measurements were
carried out by the medical
staff according to the
standard protocols of
WHO. Height was
measured using a
mechanical height gauge
to the nearest 0.1 cm.
Weight was measured
using digital scales to the
nearest 100 g.

Age at outcome:

7-11 yr

Weight (kg): -1.96 (-3.11,
-0.82)

Bust (cm): -2.77 (-3.79, -1.76)

Waistline (cm): -3.65 (-4.78,
-2.52)

BMI (kg/m2): -0.20 (-0.65,

0.25)

Zhou et al. (2020)

Taizhou
China

April 2013-
November 2013

Cross-sectional

n: 1,678

Children 6 yr or older

Blood

Blood was measured by
GFAAS

Age at Measurement:
>6 yr

Meanb: 5.684 pg/dL

Geometric meanb:
4.904 pg/dL

Medianb: 4.644 pg/dL
75thb: 6.4 pg/dL
Maxb: 46.8 pg/dL

Tertilesb (pg/dL)

Postnatal growth: HAZ,
WAZ and BMIZ

Children's body weight
and supine length or
standing height were
measured. BMI was
calculated by the formula
BMI = weight (kg)/height
(m)2; Z-scores of
anthropometric
parameters, such as HAZ,
WAZ and BMIZ, were
calculated with the WHO
Child Growth Standards.

Multivariable linear
models were adjusted
for age, sex, BW,
maternal education

(3 (95% Cl)c:

WAZ: -0.33 (-0.56, -0.11)
HAZ: -0.38 (-0.63, -0.14)
BMIZ: -0.13 (-0.37, 0.12)

WAZ Tertiles

T1
T2
T3

Reference

-0.28 (-0.47, -0.09)
-0.42 (-0.62, -0.23)

HAZ Tertiles

T1
T2
T3

Reference

-0.26 (-0.47, -0.04)
-0.36 (-0.58, -0.15)

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T1: <2.5
T2: 2.5-5.0
T3: >5.0

Age at outcome:
>6 yr



BMIZ Tertiles
T1: Reference

T2
T3

-0.18 (-0.39, 0.04)
-0.29 (-0.50, -0.07)

Males:

WAZ: -0.36 (-0.67, -0.06)
HAZ: -0.38 (-0.72, -0.04)
BMIZ: -0.15 (-0.49, 0.19)

WAZ Tertiles:

T1
T2
T3

Reference

-0.42 (-0.71, -0.13)
-0.52 (-0.81, -0.24)

HAZ Tertiles:

T1
T2
T3

Reference

-0.36 (-0.69, -0.004)
-0.43 (-0.75, -0.11)

BMIZ Tertiles:

T1: Reference

T2: -0.28 (-0.60, 0.04)

T3: -0.35 (-0.68, -0.03)

Females

WAZ: -0.29 (-0.61, 0.03)
HAZ: -0.35 (-0.71, 0.01)
BMIZ: -0.10 (-0.45, 0.26)

WAZ Tertiles:

T1: Reference

T2: -0.17 (-0.42, 0.09)

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T3

-0.36 (-0.62, -0.09)

HAZ Tertiles:

T1

Reference

T2

-0.17 (-0.45, 0.11)

T3

-0.31 (-0.60, -0.02)

BMIZ Tertiles:

T1

Reference

T2

-0.10 (-0.38, 0.18)

T3

-0.25 (-0.54, 0.04)

Choi eta!. (2017)
Seoul

South Korea

July 2014 to June
2016

Cross-sectional

n: 210

Children ranging from 8
to 23 mo in age and
healthy; no intake of
herbal medicine, iron, or
zinc supplements in the
past 3 mo; no acute
febrile disease or acute
gastrointestinal disease
in the past 2 wk; and no
evidence of other acute
or chronic diseases
affecting growth on
physical examination or
in medical history

Blood

Blood was measured by
ICP-MS

Age at Measurement:
8-23 mo

Geometric mean:
0.96 [jg/dL Median:
0.83 [jg/dL
75th: 1.23 [jg/dL
Max: 3.5 [jg/dL

Postnatal growth: Weight,
height, HC

Each infant's weight,
height, and HC were
measured by experienced
nurse; iron deficiency and
iron deficiency anemia,
complete blood count,
serum iron and ferritin
concentrations, as well as
total iron-binding capacity
were measured from the
venous blood samples of
infants

Age at outcome:

8-23 mo

Linear regression
models; BW,
sociodemographic and
feeding-related factors,
and iron and anemia
status

(3 (95% CI):

WAZ-BWZ (difference of the
WAZ at the time of the study
and BWZs): -0.238 (-0.391,
-0.085)

HCAZ: -0.213 (-0.366, -0.06)

Martin et al.

Vancouver,
Edmonton,
Winnipeg, Sudbury,
Ottawa, Kingston,

MIREC Study
n: 449

MIREC study is a
national-level pregnancy
cohort of 2001 women
from 10 cities across

Blood

Blood was measured by
ICP-MS

Age at Measurement:
2-5 yr

Postnatal growth: HAZ,
WAZ, BMIZ

Child anthropometry was
performed during the
home visit and served as
a measure of growth at

Linear regression
models adjusted for
maternal education,
maternal country of
birth, age, postnatal
BMI, maternal prenatal
smoking, and paternal

(3 (95% Cl)b

HAZ

Overall:

T1: Reference

T2: -0.015 (-0.23, 0.20)

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Toronto, Hamilton,
Montreal, and
Halifax
Canada

2008-2011

Cross-sectional

Canada including
Vancouver, Edmonton,
Winnipeg, Sudbury,
Ottawa, Kingston,
Toronto, Hamilton,
Montreal, and Halifax.
Participants were
recruited in the first
trimester of pregnancy
between 2008 and 2011
and followed through
delivery.

Median: 0.663 [jg/dL
75th: 0.962 pg/dL
Max: 5.49 pg/dL

Tertiles (pg/dL)

T1
T2
T3

<0.54

0.54-0.82

>0.82

that time. Weight and
height were measured
using a calibrated scale
and calibrated
stadiometer. All
measurements were
completed in duplicate or,
if warranted due to
predefined differences in
duplicate measurements,
in triplicate.

Age at outcome:

2-5 yr

BMI; models were
additionally adjusted for
maternal metal
concentrations

T3: 0.025 (-0.20, 0.25)
Male

T1
T2
T3

Reference
0.003 (-0.28, 0.29)
-0.039 (-0.32, 0.24)

Female

T1
T2
T3

Reference
0.022 (-0.31, 0.35)
0.095 (-0.26, 0.45)

WAZ
Overall

T1
T2
T3

Reference
0.064 (-0.12, 0.25)
-0.004 (-0.20, 0.19)

Male

T1
T2
T3

Reference

0.11 (-0.15, 0.36)

0.074 (-0.18, 0.33)

Female

T1
T2
T3

Reference
0.050 (-0.22, 0.32)
-0.11 (-0.40, 0.18)

BMIZ
Overall

T1: Reference
T2: 0.097 (-0.098, 0.29)
T3: -0.041 (-0.24, 0.16)
Male

T1: Reference
T2: 0.15 (-0.13, 0.42)

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T3: 0.14 (-0.14, 0.41)
Female

T1
T2
T3

Reference
0.039 (-0.24, 0.32)
-0.26 (-0.55, 0.033)

Adjusted for maternal

exposure:

HAZ

Overall:

T1
T2
T3

Reference
-0.030 (-0.25, 0.19)
-0.008 (-0.25, 0.23)

Male

T1
T2
T3

Reference
-0.007 (-0.30, 0.28)
-0.067 (-0.38, 0.24)

Female

T1
T2
T3

Reference
0.013 (-0.33, 0.36)
0.081 (-0.30, 0.46)

WAZ
Overall

T1
T2
T3

Reference
0.041 (-0.15, 0.23)
-0.05 (-0.26, 0.16)

Male

T1
T2
T3

Reference
0.09 (-0.16, 0.35)
0.04 (-0.24, 0.32)

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Female

T1
T2
T3

Reference
0.024 (-0.26, 0.30)
-0.15 (-0.46, 0.17)

BMIZ
Overall

T1
T2
T3

Reference
0.076 (-0.12, 0.28)
-0.086 (-0.30, 0.14)

Male

T1
T2
T3

Reference
0.14 (-0.14, 0.41)
0.11 (-0.19, 0.41)

Female

T1
T2
T3

Reference

0.006 (-0.28, 0.29)

-0.32 (-0.64, 0.0036)

Jedrychowski et al.
(2015)

Krakow
Poland

January 2001-
February 2004

Cohort

Krakow Cohort Study
n: 379

The present analysis
was restricted to 379
term-babies (born
>36 wk of gestation)
who took part in the 9-yr
follow-up. Women who
were residents of
Krakow, one of the
major cities in Poland,
and attended
ambulatory prenatal
clinics in the first and
second trimesters of
pregnancy were eligible

Blood and cord blood

Maternal and UCB,
obtained at delivery, and
blood (capillary),
obtained at age 5, were
measured by high-
performance liquid
chromatography
atmospheric-pressure
ionization tandem mass
spectrometry

Age at Measurement:
Maternal age at delivery
and 5 yr old

Postnatal growth: Height
gain

At ages of 3-9 children
were invited annually for
pediatric examination
during which height
measurements were
done.

Age at outcome:
3-9 yr old

GEE models were
adjusted for maternal
height, BL, pre-
pregnancy maternal
weight, gestational
weight gain, prenatal
and postnatal ETS,
breastfeeding, maternal
education, and parity

(3 (95% CI), as mean height
growth (cm) by UCB tertiles

T1
T2
T3

Reference

-0.671 (-1.610, 0.267)
-0.736 (-1.779, 0.307)

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for the study. Enrollment
included only
nonsmoking women with
singleton pregnancies
between the ages of 18
and 35 yr who were free
from such chronic
diseases as diabetes
and hypertension.

Geometric mean:
UCB: 1.21 [jg/dL
Blood: 2.05 pg/dL

UBC Tertiles (pg/dL)

T1
T2
T3

<1.0

1.1-1.4

>1.4

Kimetal. (2017)
Korea

January 2011-
December 2012

Cohort

CHECK
n: 280

Healthy pregnant
women with mature term
singleton were recruited,
who did not have
preterm delivery,
medical predisposition,
or history of
occupational exposure

Cord blood

UCB was measured by
GFAAS

Age at measurement:
birth

Mean:

Overall: 1.31 pg/dL
Males: 1.39 pg/dL
Females: 1.21 pg/dL

Postnatal growth: Weight, Generalized linear

height, and BMI

Weight and height were
measured by the health
professionals

Age at outcome:

3, 6, 9, 12, 15, 18, 24, and

27 mo of age

model adjusted for
maternal age, maternal
BMI, gestational period,
cesarean section, and
smoking

(3 (95% Cl)b
Weight

At birth: 0.037 (-0.128, 2.01)
3 mo: -0.039 (-0.414, 0.335)
6 mo: -0.391 (-0.814, 0.033)
9 mo: 0.000 (-0.356, 0.357)
12 mo: 0.125 (-0.302, 0.552)
15 mo: 0.093 (-0.396, 0.582)
18 mo: 0.897 (-0.171, 1.965)
24 mo: 0.717 (0.195, 1.239)
27 mo: 0.316 (-0.345, 0.977)

Height

At birth: 0.176 (-0.003, 0.354)
3 mo: -0.023 (-0.384, 0.337)
6 mo: 0.033 (-0.458, 0.523)
9 mo: 0.049 (-0.346, 0.444)
12 mo: -0.058 (-0.531, 0.415)
15 mo: 0.226 (-0.220, 0.671)
18 mo: 0.909 (-0.222, 2.040)
24 mo: 0.138 (-0.530, 0.806)
27 mo: 0.354 (-0.497, 1.205)

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BMI

At birth: -0.167 (-0.357, 0.023)
3 mo: -0.019 (-0.431, 0.392)
6 mo: -0.461 (-0.937, 0.014)
9 mo: -0.031 (-0.430, 0.369)
12 mo: -0.020 (-0.492, 0.452)
15 mo: -0.098 (-0.481, 0.285)
18 mo: 0.157 (-1.266, 1.580)
24 mo: 0.695 (0.077, 1.313)
27 mo: 0.409 (-0.398, 1.216)

Males
Weight

At birth: 0.088 (-0.140, 0.316)
3 mo: -0.008 (-0.597, 0.581)
6 mo: -0.023 (-0.543, 0.497)
9 mo: 0.167 (-0.398, 0.733)
12 mo: 0.202 (-0.631, 1.034)
15 mo: 0.365 (-0.467, 1.197)
18 mo: 1.324 (0.023, 2.626)
24 mo: 0.962 (0.181, 1.743)
27 mo: 0.417 (-0.631, 1.465)
Height

At birth: 0.270 (0.037, 0.502)
3 mo: 0.232 (-0.262, 0.726)
6 mo: -0.077 (-0.695, 0.540)
9 mo: 0.166 (-0.363, 0.695)
12 mo: -0.147 (-1.153, 0.859)
15 mo: 0.433 (-0.147, 1.013)
18 mo: 1.648 (0.270, 3.026)
24 mo: 1.062 (-0.132, 2.255)

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27 mo: 1.618 (-0.450, 3.686)
BMI

At birth: -0.194 (-0.413, 0.025)
3 mo: -0.130 (-0.800, 0.540)
6 mo: 0.003 (-0.558, 0.563)
9 mo: -0.009 (-0.522, 0.504)
12 mo: 0.314 (-0.689, 1.318)
15 mo: -0.049 (-0.569, 0.470)
18 mo: 0.319 (-1.496, 2.135)
24 mo: 0.472 (-0.172, 1.116)
27 mo: 0.966 (-1.390, 3.322)

Females
Weight

At birth: 0.006 (-0.236, 0.248)
3 mo: -0.072 (-0.640, 0.496)
6 mo: -0.828 (-1.502, -0.154)
9 mo: -0.098 (-0.602, 0.407)
12 mo: 0.101 (-0.443, 0.644)
15 mo: -0.039 (-0.722, 0.643)
18 mo: -0.826 (-15.627,
13.976)

24 mo: 0.821 (-0.087, 1.728)
27 mo: 0.236 (-1.089, 1.561)
Height

At birth: 0.102 (-0.177, 0.381)
3 mo: -0.249 (-0.875, 0.378)
6 mo: 0.106 (-0.732, 0.945)
9 mo: 0.104 (-0.526, 0.734)
12 mo: -0.057 (-0.608, 0.493)
15 mo: 0.121 (-0.664, 0.905)
18 mo: -0.788d

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24 mo: -0.176 (-1.225, 0.874)
27 mo: -0.153 (-1.405, 1.100)
BMI

At birth: -0.142 (-0.474, 0.189)
3 mo: 0.098 (-0.491, 0.687)
6 mo: -0.974 (-1.778, -0.170)
9 mo: -0.143 (-0.805, 0.519)
12 mo: -0.147 (-0.688, 0.393)
15 mo: -0.103 (-0.712, 0.505)
18 mo: -2.263d
24 mo: 1.108 (-0.147, 2.362)
27 mo: 0.439 (-1.581, 2.460)

Hong et al. (2014)

Seoul, Cheonan,
and Ulsan
South Korea

May 2006 to
December 2010

Cohort

MOCEH
n: 1,751

This research was
conducted as a part of
MOCEH, which is a
multicenter prospective
hospital and community-
based birth cohort study.
Women who lived in
these cities were
enrolled in the first
trimester. The
participants fulfilled the
inclusion criterion of age
>18 yr. Written informed
consent was obtained at
the initial visit from all
enrolled mothers on
behalf of themselves
and their children. The
study subjects were
restricted to those in
which maternal and cord
BLLs were assessed,

Blood and cord blood

Maternal blood, obtained
during early pregnancy
(before gestational week
20) and at delivery, and
UCB were measured by
AAS

Age at Measurement:
maternal age at week 20
and at delivery; delivery

Mean:

Early pregnancy:
1.25 [jg/dL Late
pregnancy: 1.25 [jg/dL
UCB: 0.91 [jg/dL
Median:

Early pregnancy:
1.29 [jg/dL

Postnatal growth: weight
Z-score, length z-cores

Weights and lengths at 6
and 12 mo were taken by
using an infantometer by
laying infants on the
center of a scale and were
read to 1 decimal place
for weight (0.1 kg) and
length (0.1 cm). At 24 mo
of age, weights and
lengths were obtained by
using an automatic
measuring station for
weight and length by
standing

on the center of the scale
on both feet, and placing
their heels, bottom, back,
and posterior head on the
measuring rod.

Multivariable regression
models were adjusted
for mother's age,
education, pre-
pregnancy BMI, GA,
gender of the child, and
clinic location, and
calcium intake

(3 (95% Cl)b
Maternal Blood: Early
pregnancy Pb
Weight Z-scores
At birth: -0.05 (-0.16, 0.07)
6 mo: -0.03 (-0.19, 0.13)
12 mo: -0.10 (-0.26, 0.06)
24 mo: -0.05 (-0.23, 0.12)
Length Z-scores
At birth: 0.01 (-0.15, 0.18)
6 mo: -0.17 (-0.37, 0.02)
12 mo: 0.04 (-0.15, 0.24)
24 mo: -0.15 (-0.35, 0.04)

Maternal Blood: Late
Pregnancy Pb

Weight Z-scores

At birth: -0.01 (-0.15, 0.12)

6 mo: -0.15 (-0.34, 0.03)

12 mo: -0.15 (-0.34, 0.03)

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and postnatal growth
measurements were
performed. Exclusion
criteria: LBW(<2500 g);
preterm birth
(gestational week <37);
and missing information
on maternal age, BMI,
education level, and
gestational week;
subjects with >2 SD for
mean maternal BLLs
and child BW or length

Late pregnancy:
1.27 [jg/dL

UCB: 0.93 pg/dL

75th:

Early pregnancy:
1.65 pg/dL
Late pregnancy:
1.64 pg/dL
UCB: 1.19 pg/dL
Max:

Early pregnancy:
2.63 pg/dL

Late pregnancy:
2.52 pg/dL

UCB: 1.90 pg/dL

Age at outcome:
6, 12 and 24 mo

24 mo: -0.33 (-0.53, -0.13)
Length Z-scores
At birth: -0.07 (-0.25, 0.11)
6 mo: -0.05 (-0.28, 0.16)
12 mo: 0.10 (-0.12, 0.33)
24 mo: -0.30 (-0.53, -0.08)

UCB Pb

Weight Z-scores
At birth: 0.08 (-0.04, 0.21)
6 mo: 0.10 (-0.07, 0.28)
12 mo: 0.06 (-0.10, 0.24)
24 mo: -0.01 (-0.21, 0.18)
Length Z-scores
At birth: 0.14 (-0.03, 0.32)
6 mo: 0.11 (-0.11, 0.33)
12 mo: 0.22 (0.01, 0.44)
24 mo: 0.004 (-0.22, 0.22)

Renzefti et al.
(2017)

Mexico City
Mexico

July 2007-February
2011

Cohort

PROGRESS
n: 513

Women were
considered eligible for
enrollment if they were
18 yr or older, pregnant
at <20 wk of gestation,
free of heart or kidney
disease, did not use
steroids or anti-epilepsy
drugs, did not consume
alcohol on a daily basis,
had access to a
telephone, and planned
to reside in Mexico City
for the following 3 yr

Blood, cord blood, and
bone

Maternal blood, collected
in the second and third
trimester of pregnancy
and within 12 hr of
delivery, and UCB,
collected within 12 hr of
delivery, were measured
by ICP-QQQ.

Maternal bone,
measured at 1-mo
postpartum from tibia
(cortical bone) and
patella (trabecular bone),

Postnatal growth: HAZ,
WAZ, BMIZ, and
percentage body fat

Trained research
assistants collected
measures of

anthropometry at the age
4-6-yr visit in which child
weight and standing
height were measured
using a professional
digital scale. BMI was
calculated from height and
weight and to determine
BMIZ for age and sex
based on WHO norms.

Multivariable linear
regression adjusted for
mother's age, BMI
(height when the
outcome is HAZ),
education, GA (weeks),
primiparity, smoke
exposure, delivery
mode, breastfeeding,
sex of the child, food
frequency questionnaire
total dietary intake,
LeadCare childhood
blood Pb, and child's
age (when the outcome
is percent body fat)

(3 (95% Cl)c
HAZ

Maternal blood, second
trimester: -0.04 (-0.13, 0.04)

Maternal blood, third trimester:
-0.10 (-0.19, -0.01)

Maternal blood, at delivery:
-0.04 (-0.13, 0.05)

UCB: -0.04 (-0.14, 0.06)
Maternal patella: 0.01 (-0.003,
0.02)

Maternal tibia: -0.003 (-0.01,
0.01)

WAZ

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were measured using a
K-XRF instrument

Age at Measurement:
Maternal age at second
and third trimester and at
birth; child's age at
follow-up (4-6 yr)

Mean (SD):

Maternal blood - second
trimester: 3.7 (2.6) [jg/dL
Maternal blood - third
trimester: 3.9 (2.8) [jg/dL

Maternal blood - at
delivery: 4.3 (3.1) [jg/dL

UCB: 3.5 (2.7) pg/dL

Patella: 4.7 (8.8) pg/g

Tibia: 2.9 (8.6) pg/g

Geometric mean:
Maternal blood - second
trimester: 3.0 pg/dL
Maternal blood - third
trimester: 3.1 pg/dL

Maternal blood - at
delivery: 3.5 pg/dL

UCB: 2.8 pg/dL
Max:

Maternal blood - second
trimester: 17.8 pg/dL

Maternal blood - third
trimester: 28.3 pg/dL
Maternal blood - at
delivery: 21.9 pg/dL

Tetrapolar bioelectrical
impedance was measured
to estimate body fat mass
and percent body fat

Age at outcome:

4-6 yr old

Maternal blood, second
trimester: -0.02 (-0.13, 0.09)

Maternal blood, third trimester:
-0.11 (-0.22, -0.003)

Maternal blood, at delivery:
-0.03 (-0.13, 0.08)

UCB: -0.03 (-0.15, 0.09)

Maternal patella: 0.01 (-0.01,
0.02)

Maternal tibia: -0.0003 (-0.01,
0.01)

BMIZ

Maternal blood, second
trimester: 0.04 (-0.07, 0.15)

Maternal blood, third trimester:
-0.01 (-0.12, 0.10)

Maternal blood, at delivery:
-0.03 (-0.08, 0.14)

UCB: 0.05 (-0.08, 0.17)

Maternal patella: 0.01 (0.01,
0.02)

Maternal tibia: 0.01 (-0.01,
0.02)

Percentage of body fat

Maternal blood, second
trimester: -0.13 (-0.75, 0.49)

Maternal blood, third trimester:
-0.21 (-0.82, 0.41)

Maternal blood, at delivery:
-0.12 (-0.74, 0.50)

UCB: 0.31 (-0.37, 0.99)

Maternal patella: 0.01 (-0.06,
0.07)

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Study Design

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Outcome

Confounders

Effect Estimates and 95%
Cisa

UCB: 18.5 [jg/dL
Patella: 43.2 |jg/g
Tibia: 30.1 |jg/g

Maternal tibia: 0.01 (-0.06,
0.08)

Liu et al. (2019a)

Mexico City
Mexico

1994-2003

Cohort

ELEMENT
n: 248

Pregnant women who
were recruited from
three maternity hospitals
in Mexico City and
followed for 12 mo post-
partum and children
followed through age 4

Blood and bone

Maternal tibia (cortical)
and patella (trabecular)
bone, measured at 1-mo
postpartum, were
measured using a
noninvasive spot-source
Cd K-XRF instrument
constructed at Harvard
University. Blood,
obtained from each child
annually from 1 to 4 yr,
was measured by
GFAAS

Age at Measurement:
maternal age at delivery
and 1-4 yr old

Mean:

Maternal patella:
12.3 |jg/g

Maternal tibia: 8.9 |jg/g

Blood (cumulative):
19.6 [jg/dL

Median:

Maternal patella:

10.6	|jg/g

Maternal tibia: 8.3 |jg/g

Blood (cumulative):

17.7	[jg/dL
75th:

Postnatal growth: BMIZ,
WC, sum of skinfolds, and
body fat percentage

At the follow-up visit, child
weight, height, WC, and
skinfold thickness (biceps,
subscapular and
suprailiac) were measured

Age at outcome:
8-16 yr old

Multivariable linear
regression models were
adjusted for maternal
age, parity, education
and calcium treatment
group, and children's
age, sex, and pubertal
stage

(3 (95% Cl)c
BMIZ

Patella: -0.02 (-0.03, -0.01)
Tibia: -0.00 (-0.02, 0.01)
Blood: 0.02 (-0.40, 0.45)

WC (cm)

Patella: -0.12 (-0.22, -0.03)
Tibia: -0.07 (-0.21, 0.07)
Blood: -0.38 (-3.74, 2.97)

Sum of skinfolds (mm)
Patella: -0.29 (-0.50, -0.08)
Tibia: -0.10 (-0.38, 0.19)
Blood: -1.62 (-8.76, 5.52)

Body of fat percentage (%)
Patella: -0.09 (-0.17, -0.01)
Tibia: -0.01 (-0.13, 0.10)
Blood: 2.08 (-0.98, 5.13)

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Study Design

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Outcome

Confounders

Effect Estimates and 95%
Cisa

Maternal patella:
19.7 |jg/g

Maternal tibia: 15.2 |jg/g

Blood (cumulative):
23.5 [jg/dL
Max:

Maternal patella:
50.1 |jg/g

Maternal tibia: 38.6 |jg/g
Blood (cumulative):
55.0 [jg/dL

Afeiche et al. (2012) n: 773

Mexico City
Mexico

1994-2005

Cohort

Mothers were recruited
from maternity hospitals
serving low-to-moderate
income populations in
Mexico City; preterm
(<37 wk) and LBW
(<2500 g) were
excluded

Blood and bone

Maternal bone, assessed
at approximately 1 mo
postpartum, measured
by in vivo K-XRF from
the mid-tibial shaft
(cortical bone) and the
patella (trabecular bone);
blood, obtained from
children at 24 mo or 30-
48 mo, was measured by
GFAAS.

Age at Measurement:
Maternal age 1 mo
postpartum, with average
age at delivery: 25.7 (SD:
5.3) yr; birth—24 mo; 30-
48 mo

Postnatal growth: Attained
height and BMI

Children's weight and
height were measured
and recorded by trained
staff members at birth and
age 48 mo using standard
protocols

Age at outcome:
birth and 48 mo

Linear regression
models were adjusted
for maternal height and
calf circumference,
number of previous
pregnancies, marital
status, education level,
breastfeeding for 6 mo,
cohort, calcium
treatment group
assignment during
lactation and
pregnancy, age at
delivery, and child sex
and GA at birth; all
height models were
additionally adjusted for
BL; BMI models were
additionally adjusted for
BW

(3 (95% CI)

Height differences (cm)

Prenatal: -4.6 (-10.25, 1.05)

Infant blood: -0.84 (-1.43,
-0.26)

Childhood blood: 0.41 (-0.17,
0.99)

BMI difference (kg/m2)
Prenatal: -0.70 (-3.05, 1.65)
Infant blood: -0.07 (-0.32,
0.18)

Childhood blood: 0.09 (-0.15,
0.33)

Median:

Maternal tibia: 8.2 |jg/g
Maternal patella: 9.4 |jg/g
Infant blood (average

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Outcome

Confounders

Effect Estimates and 95%
Cisa

from birth to 24 mo)
4.5 [jg/dL

Childhood blood
(average from 30-
48 mo): 5.6 [jg/dL

Kerr et al. (2019)

Torreon
Mexico

February 2001-
June 2002

Cohort

n: 538

Children attending nine
public elementary
schools located within a
3.5 km radius from a
foundry close to the city
center participated in the
study. Participants were
randomized into one of
four groups: iron (30 mg
of ferrous fumarate),
zinc (30 mg zinc oxide),
a combination of iron
and zinc or a placebo
(sugar pill)

Blood

Blood, collected at
baseline (T1), 6 mo after
baseline (T2), and 12 mo
after baseline (T3), was
measured by GFAAS

Age at Measurement:
6-8 yr old

Median: 10.1 pg/dl_
75th: 23.7 pg/dL

Postnatal growth: height,
knee height, and HAZ

A single trained individual
took anthropometric
measures at each time
point (T1, T2, T3),
according to standard
methods recommended
by the WHO; Height and
knee height were
measured without shoes
using a standardized
measuring board or a
knemometer, respectively,
to the nearest 1 mm;

Age at outcome:

6-8 yr old

Multivariable linear
regression adjusted for
age, sex, mother's
education, crowding,
and hemoglobin at
baseline; HAZ models
were not adjusted for
age or sex; models
were also stratified by
ALAD genotype

(3 (95% CI)

Height: -0.11 cm ( -0.18,
-0.04)

Knee height: -0.04 cm (-0.07,
-0.02)

HAZ: -0.02 cm ( -0.03, -0.01)

ALADl-2/2-2

Height: -0.38 cm (-0.68, 0.09)

Knee height: -0.14 cm (-0.25,
-0.02)

HAZ: -0.07 (-0.12, -0.02)
ALAD1-1

Height: -0.09 cm (-0.16,
-0.02)

Knee height: -0.04 cm (-0.06,
-0.01)

HAZ: -0.02 (-0.03, -0.004)

Burns etal. (2017)

Chapaevsk
Russia

2003-2005 (2012—
2015)

Cohort

Russian Children's

Study

n: 499

The Russian Children's
Study is a prospective
cohort of 499 boys
residing in Chapaevsk,
Russia, enrolled in
2003-2005 at ages 8-
9 yr and followed

Blood

Blood measured by
GFAAS with Zeeman
background corrected

Age at Measurement:
8-9 at enrollment

Postnatal growth: HAZ
and BMIZ

At study entry and annual
follow-up visits, a
standardized
anthropometric
examination was
performed according to a
written protocol. Height
was measured to the

Mixed effects linear
regression models were
adjusted for BW,
preterm birth, percent
calories from protein at
baseline, and age for
the HAZ models and
BW, no biological father
in home, percent
calories from fat at

(3 (95% CI)b, as estimated
mean growth Z-scores
comparing higher (>5 pg/dL) to
lower (<5 pg/dL) BLL
HAZ: -0.43 (-0.60, -0.25)
BMIZ: -0.22 (-0.45, 0.006)

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Cisa

annually through 2012-
2015 to age
18 yr. For this analysis,
10 boys in the original
cohort were excluded
due to chronic illnesses
that could affect growth
and/or pubertal
development.

Median: 3.0 [jg/dL
Max: 31 [jg/dL

nearest 0.1 cm using a
stadiometer. Weight was
measured to the nearest
100 g with a metric scale.
HAZ and BMIZ were
calculated using the WHO
standards

Age at outcome:
8-9 at enrollment and
annually through age 18

baseline, and age for
BMIZ models

Deierlein et al.

New York City, NY;
Cincinnati, OH; and
San Francisco, CA
United States

2004-2007

Cohort

Breast Cancer and
Environment Research
Program
n: 683

Girls ages 6-8 yr were
enrolled in 2004-2007
at three sites: New York
City, Cincinnati, and San
Francisco; girls have no
underlying endocrine
medical conditions, be
of Black or Hispanic
race/ethnicity (New York
City site only), and have
been born in the Kaiser
Permanente system
(san Francisco)

Blood

Blood was measured by
ICP-MS

Age at Measurement:
6-10 yr

Median: 0.99 [jg/dL

Mean (SD): 1.16
(0.67) [jg/dL
Geometric mean:
1.03 [jg/dL (95% CI:
0.99, 1.07)

Max: 5.40 [jg/dL

Postnatal growth: height,
BMI, WC, and percent
body fat

Weight (kg), standing
height (cm), and umbilical
WC (cm) were collected at
baseline and at biannual
(Cincinnati) or annual
(New York City and San
Francisco Bay Area)
follow-up visits by trained
interviewers using a
standard protocol; BMI
was calculated as weight
divided by squared height
(kg/m2). Percent body fat
was estimated using
bioelectrical impedance
analysis

Age at outcome:

7-14 yr

Linear mixed effects
models with an
unstructured correlation
matrix were adjusted for
age, age squared, race,
an Interaction term
between age and blood
Pb concentrations, an
interaction term
between age squared
and blood Pb
concentrations, and an
interaction term
between race and age

(3 (95% CI)





Height (cm)





Age

7:

-2.0 (-

-3.0,

-1.0)

Age

8:

-1.9 (-

-2.8,

-0.9)

Age

9:

-1.7 (-

-2.7,

-0.8)

Age

10

-1.6

(-2.6

-0.7)

Age

11

-1.6

(-2.5,

-0.6)

Age

12

-1.5

(-2.5,

-0.5)

Age

13

-1.5

(-2.5,

-0.5)

Age

14

-1.5

(-2.5,

-0.4)

BMI

(kg/m2)





Age

7:

-0.7 (-

-1.2,

-0.2)

Age

8:

-0.8 (-

-1.3,

-0.3)

Age

9:

-0.9 (-

-1.4,

-0.4)

Age

10

-0.9

(-1.4,

-0.4)

Age

11

-0.9

("1.5,

-0.3)

Age

12

-0.9

("1.5,

-0.3)

Age

13

-0.8

("1.5,

-0.2)

Age

14

-0.8

("1.5,

-0.02)

WC (cm)





Age 7:

-2.2 (-

-3.8,

-0.6)

Age

8:

"2.5 (-

-3.8,

-1.1)

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Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Cisa

Age 9:
Age 10
Age 11
Age 12
Age 13
Age 14
Percent
Age 7
Age 8
Age 9
Age 10
Age 11
Age 12
Age 13
Age 14

¦2.7 (-4.0, -1.4)
-2.9 (-4.9, 1.4)
-3.0 (-4.5, -1.4)
-3.0 (-4.7, -1.3)
-3.0 (-4.8, -1.1)
-2.9 (-4.8, -0.9)
body fat (%)
¦1.8 (-3.2, -0.4)
¦2.0 (-3.3, -0.7)
¦2.1 (-3.4, -0.8)
-2.2 (-3.4, -0.9)
-2.1 (-3.4, -0.9)
-2.1 (-3.4, -0.8)
-1.9 (-3.2, -0.6)
-1.7 (-3.1, -0.4)

Raihan et al. (2018) MAL-ED study
n: 729

Mirpur, Dhaka
Bangladesh

Blood

Blood was measured

Children under the age
of2	a using GFAAS

November 2009-
December 2012

Cross-sectional

Age at measurement:
under the age of 2

Mean: 8.25 [jg/dL

Postnatal growth:
Stunting, wasting,
underweight

Child's length and weight
were measured using
Seca 417 infantometer
(precision: ± 1 mm) and
Seca 354 Dual Purpose
Baby Scale (precision:
10 gm).

Age at outcome:
under the age of 2

Logistic regression
models were adjusted
for child's gender,
weight, maternal
education, BMI, average
household income and
HFIAS categories in
stunting models; child's
gender, age, maternal
education, BMI, average
household income and
HFIAS categories in the
wasting models; and
child's gender, length,
maternal education,
BMI, average household
income and HFIAS
categories in the
underweight models

OR (95% CI)

Stunting: 1.78 (1.07, 2.99)
Wasting: 1.18 (0.64, 2.19)
Underweight: 1.63 (1.02, 2.61)

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Cisa

Gleason et al.

Sirajdikhan and
Pabna Upazilas
Bangladesh

2008-2011 (2010-
2013)

Cohort

n: 618

Children of mother's
from Sirajdikhan and
Pabna Upazilas of
Bangladesh between
2008 and 2011;

Between 2010 and
2013, when children
were aged 12 to 40 mo,
healthcare workers from
Dhaka Community
Hospital invited families
to enroll their children in
follow-up studies

Cord blood

UCB were measured by
ICP-MS and child's
blood, collected at 20 to
40 mo, was measured by
portable LeadCare II
instruments

Age at Measurement:
at birth and 12-40 mo

Median:

UCB: 3.1 [jg/dL

Blood: 4.2 [jg/dL

75th:

UCB: 6.3 [jg/dL
Blood: 7.6 [jg/dL

Postnatal growth: Stunting

Stunting status of children
was determined using the
WHO macros (Version
3.2.2)

Age at outcome:

12-40 mo

Logistic regression
models were adjusted
for maternal weight,
maternal education,
maternal protein intake,
and HOME Inventory
score were all modeled
as continuous variables;
average water As and
Mn levels were included
as continuous variables

OR (95% CI)

UCB: 0.97 (0.93, 1.00)

Blood at 20-40 mo: 1.15 (1.00,
1.33)

ALAD = 6-aminolevulinic acid dehydratase; BL = birth length; BMI = body mass index; BMIZ = BMI-for-age Z-score; BW = birth weight; BWZ = birth weight Z-score;

CHECK = Children's Health and Environmental Chemicals in Korea; CI = confidence interval; ELEMENT = Early Life Exposure in Mexico to Environmental Toxicants;
ETS = environmental tobacco smoke; GEE = generalized estimating equation; GFAAS = graphite furnace atomic absorption spectrometry; HAZ = height-for-age Z-score;

HCAZ = head circumference for age Z-score; HFIAS = Household Food Insecurity Access Scale; HOME = Health Outcomes and Measures of the Environment; hr = hour(s); ICP-
MS = inductively coupled plasma mass spectrometry; ICP-QQQ = inductively coupled plasma triple quad; K-XRF = K-shell X-ray fluorescence; LBW = low birth weight; MAL-
ED = Interactions of Malnutrition and Enteric Infections: Consequences for Child Health and Development; MIREC = Maternal-Infant Research on Environmental Chemicals;
mo = month(s); MOCEH = Mothers' and Children's Environmental Health; NHANES = National Health and Nutrition Examination Survey; OR = odds ratio; PIR = poverty-income
ratio; PROGRESS = Programming Research in Obesity, Growth, Environment and Social Stressors; SD = standard deviation; T# = fertile #; UCB = umbilical cord blood;
WAZ = weight for age Z-score; WC = waist circumference; WHO = World Health Organization; wk = week(s); yr = year(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect

estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated

interval. Categorical effect estimates are not standardized.

bPb measurements were converted from |jg/L to |jg/dL.

°Effect estimates unable to be standardized.

dNo CI reported.

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Table 8-11

Animal toxicological studies of Pb exposure and development

Study

Species (Stock/Strain), n, Sex

Timing of Exposure

Exposure
Details

(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Graham et al. (2011)

Rat (Sprague-Dawley)

Control (vehicle), M/F, n = 14-16 (7-8/7-8)

1 mg/kg Pb, M/F, n = 14-16 (7-8/7-8)
10 mg/kg Pb, M/F, n = 14-16 (7-8/7-8)

PND 4 to 28

Offspring were
dosed via gavage
every other day
from PND 4 until
PND 28.

PND 29
0.267 [jg/dL for
control

3.27 [jg/dL for
1 mg/kg

12.5 [jg/dL for
10 mg/kg

Offspring Body
Weight

de Fiqueiredo et al.
(2014)

Rat (Wistar)

28 d old Control (untreated), M,
n = 10

60 d old Control (untreated), M, n = 12

28 d old 30 mg/L Pb, M, n = 10

60 d old Control (assumed untreated), M,
n = 12

60 d old 30 mg/L Pb, M, n = 17

PND 0 to PND 28 or
PND 0 to PND 60

Male Wistar rats PND 28

Offspring Body

were dosed via
drinking water
from birth to
PND 28 or 60.

1.2 pg/dL for control Weight
8.0 pg/dL 30 mg/L Pb

PND 60

1.6 pg/dL for control
7.2 pg/dL for 30 mg/L
Pb

Duan etal. (2017) Mouse (CD-11

PND 1 to PND 21

Dams were dosed

Pups:

Offspring Body

Dams



via drinking water

PND 1

Weight

Control (0 ppm Pb), F, n = 3



starting on GD 1
and continued

1.29 pg/dL for control



Low dose (27 ppm Pb), F, n = 3



through weaning
(PND 21).

1.29 pg/dL for low
dose



High dose (109 ppm Pb), F, n = 3





1.29 pg/dL for high
dose



Pups





PND 18

1.62 pg/dL for control



Control (0 ppm Pb), NR, n = 9







Low dose (27 ppm Pb), NR, n = 9





19.6 pg/dL for low
dose



8-184


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Exposure

Study	Species (Stock/Strain), n, Sex	Timing of Exposure	BLL	Ixamined

Duration)

29.16 [jg/dL for high

High dose (109 ppm Pb), NR, n = 9	dose

PND 35

1.51 |jg/dL for control
28.7 [jg/dL for low
dose

38.0 [jg/dL for high
dose

Betharia and Maher
(2012)

Rat (Sprague-Dawley)

Dams

Control (untreated), F, n = 6
10 |jg/mL Pb, F, n = 6

Pups

Control (untreated), M/F, n = 36-48 (18—
24/18-24)

10 |jg/mL Pb, M/F, n = 36^8 (1824/18-24)

GD Oto PND 20

Dams dosed via
drinking water
starting on GD 0
through weaning
(PND 20).

Pups:

PND 2

0.188 [jg/dL for
control

9.03 [jg/dL for
10 |jg/ml_ Pb

PND 25

0.0880 [jg/dL for
0 |jg/mL
0.976 [jg/dL for
10 |jg/ml_ Pb

Offspring Body
Weight

PND 60

0.0244 [jg/dL for
control

0.0318 [jg/dL for
10 |jg/ml_ Pb

Zhaoetal. (2021)

Rat (Sprague-Dawley)

Control (untreated), F, n = 6 dams

109 ppm Pb, F, n = 6 dams

GD -14 to PND 10

Dams were dosed	Pups:
via drinking water g

starting 2 wk prior	„	,

to mating and	0.87 pg/dL for control

continued until	48.2 [jg/dL for

PND 10.	109 ppm Pb

Offspring Body
Weight

8-185


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Exposure

Study	Species (Stock/Strain), n, Sex	Timing of Exposure	BLL	Ixamined

Duration)

PND 10

0.87 [jg/dL for control
11.5 [jg/dL for
109 ppm Pb

PND 21

0.87 [jg/dL for control

2.81 [jg/dL for
109 ppm Pb

PND 30

0.87 [jg/dL for control

1.20 [jg/dL for
109 ppm Pb

Rao Barkur and Bairv

Rat (Wistar)

Control (untreated), F, n = 6 dams

0.2% Pb Pregestation Only, n = 6 dams

0.2% Pb Gestation Only, n = 6 dams

0.2% Pb Lactation Only, n = 6 dams

0.2% Pb Gestation and Lactation, F, n = 6
dams

GD -30 to GD -1; GD 0
to GD 21; PND 1 to
PND 21; GD 0 to PND 21

Dams were dosed
via drinking water
for varying
amounts of time:
Pregestation Only
(1 mo prior to
conception),
Gestation Only
(21 d), Lactation
Only (21 d), and
Gestation and
Lactation (42 d).

Pups (PND 22):
0.19 [jg/dL for control
3.03 [jg/dL for 0.2%
Pb in Pregestation
Only group
5.51 [jg/dL for 0.2%
Pb in Gestation Only
group

26.86 [jg/dL for 0.2%
Pb in Lactation Only
group

31.59 [jg/dL for 0.2%
Pb in Gestation and
Lactation group

Offspring Body
Weight, Pinna
Detachment, Eye
Opening, Tooth
Eruption

jarkur and Bairv

Rat (Wistar)

Control (untreated),

F, n = 6 dams

GD -30 to GD -1, or
GD 0 to 21, or PND 0 to
21, or GD 0 to PND 21

Dams were dosed
via drinking water
for varying
amounts of time:

Pups (PND 22):
0.18 [jg/dL for control

Offspring Body
Weight

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Study

Species (Stock/Strain), n, Sex

Timing of Exposure

Exposure
Details

(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

0.2% Pb Pregestation Only, n = 6 dams

Pregestation Only



(1 mo prior to

0.2% Pb Gestation Only, n = 6 dams

conception),



Gestation Only

0.2% Pb Lactation Only, n = 6 dams

(21 d), Lactation



Only (21 d), and

0.2% Pb Gestation and Lactation, F, n = 6

Gestation and

dams

Lactation (42 d).

3.02 [jg/dL for 0.2%
Pb in Pregestation
Only group

5.30 [jg/dL for 0.2%
Pb Gestation Only
group

26.7 [jg/dL for 0.2%
Pb in Lactation Only
group

32.0 [jg/dL for 0.2%
Pb in Gestation and
Lactation group

Sobolewski et al.

Mouse (C57BL/6)

Control (untreated) F, n = 10,

100 ppm Pb, F, n = 10

GD -
only

61 to PND 21 of F1

Dams were dosed
via drinking water
beginning 2 mo
prior to breeding
and ending on
PND 21 of the F1
(weaning).

F1

PND 6-7
0.0 [jg/dL for control,
12.5 [jg/dL for
100 ppm Pb

F3

Postnatal Mo 6-7

0.0 [jg/dL for control,
0.4 |jg/dL for 100 ppm
Pb

Offspring Body
Weight

Albores-Garcia et al.
(2021)

Rat (Long-Evans)

Evaluated on PND 14

Controls (untreated), F, n = 11 dams

Controls (untreated), M/F, n = 14 (7/7) pups

1500 ppm Pb, F, n = 7 dams

1500 ppm Pb, M/F, n = 13 (6/7) pups

Evaluated on PND 28

Controls (untreated), F, n = 9 dams

Continuous exposure
starting at GD -10

Dams were dosed
via the diet
starting 10 d prior
to mating. After
weaning
(PND 21),
offspring were put
onto the same
diet as their
dams.

Pups

PND 14

<1.9 [jg/dL for control
males

<1.9 [jg/dL for control
females

36.1 [jg/dL for
1500 ppm Pb males

37 [jg/dL for

1500 ppm Pb females

Offspring Body
Weight

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Study

Species (Stock/Strain), n, Sex

Timing of Exposure

Exposure
Details

(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Controls (untreated), M/F, n = 16 (8/8) pups

1500 ppm Pb, F, n = 8 dams

1500 ppm Pb, M/F, n = 13 (7/6) pups

Evaluated on PND 50
Controls (untreated), F, n = 15 dams
Controls (untreated), M/F, n = 15 (7/8) pups
1500 ppm Pb, F, n = 14 dams
1500 ppm, M/F, n = 15 (7/6) pups

PND 28

<1.9 [jg/dL for control
males

<1.9 [jg/dL for control
females

21.1 [jg/dL for
1500 ppm Pb males
20.9 [jg/dL for
1500 ppm Pb females

Evaluated on PND 120

Controls (untreated), F, n = 13 dams

Control (untreated), M/F, n = 13 (7/6) pups

1500 ppm Pb, F, n = 9 dams

1500 ppm Pb, M/F, n = 12 (6/6) pups

PND 50

<1.9 [jg/dL for control
males

<1.9 [jg/dL for control
females
20.2 [jg/dL for
1500 ppm Pb males

22.1 [jg/dL for
1500 ppm Pb females

PND 120

<1.9 [jg/dL for control
males

<1.9 [jg/dL for control
females

19.6 [jg/dL for
1500 ppm Pb males

24.3 [jg/dL for
1500 ppm Pb females

asaen and Sobin Mouse (C57BL/6)	PND 0 to PND 28	Dams were dosed PND 28	Offspring Body

I	Control (untreated), M/F, n = 12 (6/6)	via drinking water o.03 ug/dL for control Weight

from birth of ma|es

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Study

Species (Stock/Strain), n, Sex

Timing of Exposure

Exposure
Details

(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

30 ppm Pb, M/F, n = 12 (6/6)
330 ppm Pb, M/F, n = 12 (6/6)

offspring until
PND28.

0.03 [jg/dL for control
females

3.63 [jg/dL for 30 ppm
Pb males

2.74 |jg/dL for 30 ppm
Pb females

16.02 [jg/dL for
330 ppm Pb males

13.35 [jg/dL for
330 ppm Pb females

jarkuret al. (2011)

Rat (Wistar)

Control (untreated), F, n = 6 dams
0.2% Pb, F, n = 6 dams

GD 1 to PND 21

Dams were dosed
via drinking water
from GD 1 to
PND 21. Only
male pups were
retained for
measurements of
body weight.

Pups (males only):

PND 22

0.266 [jg/dL for
control

31.2 [jg/dL for 0.2%
Pb

PND 120
0.234 [jg/dL for
control

Offspring Body
Weight

0.468 [jg/dL for 0.2%
Pb

3asha and Reddy

Rat (Wistar)

Control (untreated), F, n = 8 dams
0.2% Pb, F, n = 8 dams

GD 6 to 21

Dams were dosed
via drinking water
from GD 6 to
PND 21. Only
male pups were
retained for
measurements of
body weight and

Pups (males only):
PND 21

0.21 [jg/dL for control

11.2 pg/dL for 0.2%
Pb

PND 28

Pinna

Detachment,
Tooth Eruption,
Fur

Development,
Eye Slit

Formation, Eye
Opening,

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Study

Species (Stock/Strain), n, Sex

Timing of Exposure

Exposure
Details

(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

developmental
milestones.

0.33 [jg/dL for control
12.3 [jg/dL for 0.2%
Pb

Offspring Body
Weight, Offspring
Body Size

Postnatal Mo 4
0.19 [jg/dL for control
5.9 |jg/dL for 0.2% Pb

BLL = blood lead level; d = day(s); GD = gestational day; F = female; M = male; mo = month(s); NR = not reported; Pb = lead; PND = postnatal day.

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Table 8-12

Epidemiologic studies of exposure to Pb and puberty in females and puberty in males

Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Effects on Puberty in Females

Yao et al. (2019)
United States
2011-2012
Cross-sectional

NHANES

n: 426 female children,
and 470 female
adolescents

Female children (age
6-11 yr) and female
adolescents (age 12-
19 yr) in NHANES
2011-2012

Blood

Blood was measured by ICP-
MS

Age at Measurement:
6-19 yr old

Geometric mean:

Female children: 0.68 [jg/dL

Female adolescents:
0.47 [jg/dL

Median:

Female children: 0.65 [jg/dL

Female adolescents:
0.47 [jg/dL

75th:

Female children: 0.93 [jg/dL

Female adolescents:
0.63 [jg/dL

Quartiles (|jg/dL):

Female children:

Q1: <0.48
Q2: 0.48-0.65
Q3: 0.65-0.93
Q4: >0.93

Female adolescents:
Q1: <0.35
Q2: 0.35-0.47

Puberty among females:
Serum tT levels

Serum tT levels were
analyzed by isotope-
dilution liquid
chromatography-tandem
mass spectrometry

Age at outcome:
6-19 yr old

Weighted multivariable
linear regression
models; Model 1
controlled forage, race,
and BMI. Model 2
controlled for PIR,
seasons of collection,
times of venipuncture,
and serum cotinine, in
addition to the
covariates of model 1

(3 (95% CI), as percent
difference in serum tT
Model 1:

Female children
Q1: Reference
Q2: 14.34 (-3.75, 35.81)
Q3: -5.00 (-21.05, 14.32)
Q4: -5.73 (-23.13, 15.61)
p for trend: 0.36
Female adolescents

Q1
Q2
Q3
Q4

Reference
-8.55 (-18.52, 2.63)
-1.95 (-13.04, 10.56)
13.12 (0.06, 27.88)

p for trend: 0.14

Model 2:

Female children
Q1: Reference
Q2: 14.9 (-3.54, 36.86)
Q3: -0.96 (-17.80, 19.34)
Q4: -2.40 (-21.00, 20.57)
p for trend: 0.63
Female adolescents

Q1
Q2
Q3
Q4

Reference
-7.83 (-18.22, 3.88)
-1.07 (-12.67, 12.06)
14.85 (0.83, 30.81)

p for trend: 0.08

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Outcome

Confounders

Effect Estimates and 95%
Clsa

Reference and
Study Design

Study Population

Exposure Assessment

Q3: 0.47-0.63
Q4: >0.63

Stawinska et al.

Legnica-Glogow

District

Poland

1995-2007

Cross-sectional

1995 n:436; 2007
n:346

Menarche status of
schoolgirls 7-16 yr
from villages in
southwestern Poland
was surveyed in 1995,
2001, 2004, and 2007.

Blood

Blood was measured by
GFAAS with a Zeeman
correction for background

Age at Measurement:
7-16 yr old

Mean

1995: 6.57 pg/dL
2007: 4.24 pg/dL

Puberty among females:
Short-term secular
change in menarche

Menarche through survey

Age at outcome:

7-16 yr

Logistic regression
models were adjusted
for age, height (linear
growth), BMI (weight-
for-height), and Pb
group (low Pb group:
2-5 pg/dL; high Pb
group: 5.10-
33.90 pg/dL)

OR (95% CI)
1995: 0.70 (0.27,
2007: 0.31 (0.09,

1.85)
1.06)

OR (95% CI)

Model with BMI: 0.54 (0.26,
1.13)

Model with percent body fat:
0.52 (0.25, 1.08)

Model with sum of skinfolds:
0.53 (0.26, 1.10)

Cross-sectional	010

Mean

Total: 3.6 pg/dL
<3.7 pg/dL: 2.9 pg/dL
>3.7 pg/dL: 4.4 pg/dL

Median

Total: 3.6 pg/dL
<3.7 pg/dL: 2.8 pg/dL
>3.7 pg/dL: 4.3 pg/dL

Gomula et al. (2022) n: 490

Polkowice
Poland

2008

Girls aged 7-16 yr
who were attending
several schools in
Polkowice in 2008.

Blood

Blood was measured by AAS
with Zeeman background
correction

Age at measurement: 7-16 yr

Puberty among females:
age at menarche

Menarche through survey

Age at outcome: 7-16 yr
old

Logistic regression
models were adjusted
for age and (1) BMI; (2)
percent body fat; and
(3) sum of skinfolds

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

De Craemer et al.
(2017)

Belgium

FLEHS I: 2002-2006,
FLEHS II: 2007-
2011, and FLEHS III:
2012-2015

Cross-sectional

FLEHS I, FLEHS II
and FLEHS III
n: FLEHS I: n = 1659,
FLEHS II: n = 606, and
FLEHS III: n = 406

Adolescents aged 14-
15 yr

Blood

Blood Pb was measured by
ICP-MS

Age at Measurement:
14-15 yr old

Geometric meanb
FLEHS I: 2.13 pg/dL
FLEHS II: 1.38 pg/dL
FLEHS III: 0.926 pg/dL

Maxb

FLEHS I: 21.2 pg/dL
FLEHS II: 7.69 pg/dL
FLEHS III: 3.86 pg/dL

Puberty among females:
Hormones and sexual
maturation in adolescents

Development of breasts in
adolescent females and
pubic hair was scored
using the international
scoring criteria of
Marshall and Tanner,
where stage 1
corresponds to the start of
puberty and stage 5 to the
adult stage. Information
on menarche was
obtained through self-
assessed questionnaires.

Age at outcome:

14-15 yr old

Logistic regression
models for female pubic
hair development and
breast development
were adjusted for age
BMI, contraceptive pill
usage; linear
regression models for
age at menarche were
adjusted for age, BMI

OR (95% Cl)c

Breast development

FLEHS I: 0.798 (0.653,
0.969)

FLEHS II: 1.318 (0.936,
2.055)

FLEHS III: 1.187 (0.886,
1.627)

Pubic hair development

FLEHS I: 1.113 (0.922,
1.349)

FLEHS II: 1.322 (0.938,
2.083)

FLEHS III: 0.919 (0.677,
1.229)

(3 (95% Cl)c

Age of menarche

FLEHS I: 0.039 (-0.072,
0.15)

FLEHS II: 0.257 (0.091,
0.424)

FLEHS III: 0.126 (-0.021,
0.273)

Nkomo et al. (2018)

Johannesburg
South Africa

Cohort

BT20+ birth cohort
n: 683

Singleton births in
which the infant
resides in

Johannesburg area for
at least 6 mo after
birth; participants must
have data for BLL at
age 13 and pubertal

Blood and cord blood

UCB collected at birth and
blood at collected at age 13
were measured by AAS with
a Zeeman background
correction

Age at Measurement:
birth and age 13

Puberty among females:
Pubertal trajectory
classes

Tanner stages of pubertal
development refer to a
standard clinical method
used to describe physical
measurements of
secondary sexual
characteristics using

Multinomial logistic
regression was used to
predict pubertal growth
trajectory class based
on BLLs at age 13 yr
and cord BLLs adjusted
for ethnicity and height
at age 8

RR (95% CI)

Development of pubic hair
UCB

Blood, >5 pg/dL vs.
<5 pg/dL

Trajectory Class 1:
Reference

Trajectory Class 2: 0.45
(0.29, 0.68)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

growth trajectory
classes

Mean (SD)

UCB: 5.8 (2.1) pg/dL

Blood: 5.0 (1.9) pg/dL

Median

UCB: 6.0 [jg/dL
Blood: 4.8 [jg/dL

75th

UCB: 7.0 [jg/dL
Blood: 7.9 [jg/dL

drawings to signal stage
of pubertal development
where stage 1 signifies
lowest level of pubertal
maturation and stage 5
denotes highest level of
pubertal maturation in
girls

Age at outcome:
9-16 yr old

Trajectory Class 3: 0.55
(0.26, 1.17)

Development of breasts
Blood, >5 pg/dL vs.
<5 pg/dL

Trajectory Class 1:
Reference

Trajectory Class 2: 0.72
(0.47, 1.11)

Trajectory Class 3: 0.63
(0.42, 0.94)

Trajectory Class 4: 0.46
(0.27, 0.77)

Liu et al. (2019b)

Mexico City
Mexico

Cohort

n: 547 (283 girls and
264 boys)

Pregnant women were
recruited at three
public maternity
hospitals (Manuel Gea
Gonzalez Hospital,
Mexican Social
Security Institute and
the National Institute
of Perinatology) in
Mexico City; and
Children at age 9.8-
18.0 yr who had at
least one
measurement of
maternal bone Pb or
childhood blood Pb

Blood and bone

Maternal bone, measured at
the mid-tibial shaft (cortical
bone) and patella (trabecular
bone) was measured by K-
XRF instrument; blood
samples from children were
measured by GFAAS

Age at Measurement:
Maternal age 1-mo
postpartum; blood measured
between 1 and 4 yr

Median

Patella: 8.20 pg/g

Tibia: 7.63 pg/g

Blood, cumulative 1-4 yr:
13.83 pg/dL

75th

Puberty among females:
Pubertal stages

In girls, the stages of
pubertal development
were defined by a
pediatrician using Tanner
staging scales for the
breast maturation and
pubic hair growth.
Menarche was measured
via a self-reported
questionnaire.

Age at outcome:
9.8-18 yr

Ordinal regression
models were adjusted
for child age at visit,
maternal education and
marital status, and
number of siblings at
birth; Cox proportional
hazard regression
models were adjusted
for number of siblings
at birth, maternal
education, and marital
status

OR (95% CI), per IQR
increase in Pb

Breast development

Patella: 0.79 (0.61, 1.01)

Tibia: 1.01 (0.75, 1.36)

Blood, cumulative 1-4 yr:
0.96 (0.92, 0.99)

Pubic hair development

Patella: 0.96 (0.76, 1.22)

Tibia: 1.12 (0.84, 1.49)

Blood, cumulative 1-4 yr:
0.95 (0.92, 0.99)

HR (95% CI)

Patella

Continuous: 0.16 (0.02,
1.07)

T1: Reference
T2: 1.10 (0.76, 1.58)

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Outcome

Confounders

Effect Estimates and 95%
Clsa

T3: 0.60 (0.41, 0.88)
Tibia

Continuous: 1.11 (0.12,
9.84)

T1: Reference

T2: 1.30 (0.86, 1.96)

T3: 1.14 (0.75, 1.72)

Blood, cumulative 1-4 yr

Continuous: 0.91 (0.77,
1.08)

T1
T2
T3

Reference
0.65 (0.46, 0.91)
0.76 (0.55, 1.06)

Reference and
Study Design

Study Population

Exposure Assessment

Patella: 15.45 |jg/g
Tibia: 13.80 |jg/g

Blood, cumulative 1-4 yr:

18.76 [jg/dL

IQR

Patella: 13.57 |jg/g
Tibia: 13.30 |jg/g
Blood, cumulative 1-4 yr:
7.66 [jg/dL

Tertiles
Patella (pg/g)

T1
T2
T3

<3.9

4.0-12.9
13.0-45.3

Tibia (|jg/g)

T1: <4.6

T2: 4.7-11.3

T3: 11.4-37.3

Blood, cumulative 1-4 yr
(pg/dL)

T1
T2
T3

<12.0

12.1-16.1

16.2-51.5

Jansen et al. (2018

Mexico City
Mexico

1997-2004(2015)
Cohort

ELEMENT project
n: 200

Mothers were
recruited from prenatal
clinics ofthe Mexican
Social Security
Institute in Mexico City
who were not planning
to leave the area

Blood

Maternal blood was
measured by GFAAS

Age at Measurement:
maternal age at sampling

Median

Puberty among females:
Menarche

Girls were asked about
menarche during the
follow-up visit (between
age 9.8 and 18.1 yr).

They were asked whether
or not menarche had
occurred (Yes, no, or

Interval-censored Cox
regression models,
comparing the hazard
of menarche among
girls with prenatal
maternal blood Pb
>5 pg/dL to those with
prenatal maternal BLL
<5 pg/dL, were
adjusted for maternal

HR (95% CI)

Interval-censored Cox
models

First trimester maternal
blood

<5 pg/dL: Reference
>5 pg/dL: 0.85 (0.46, 1.24)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

within 5 yr; had a
history of infertility,
diabetes, or psychosis;
consuming alcoholic
beverages daily during
pregnancy; addiction
to illegal drugs;
diagnosis of a high-
risk pregnancy; or
being pregnant with
multiples

First trimester: 4.8 [jg/dL
Second trimester: 4.0 [jg/dL
Third trimester: 4.5 [jg/dL

75th:

First trimester: 7.1 [jg/dL
Second trimester: 6.4 [jg/dL
Third trimester: 6.6 [jg/dL

don't know/refused) and,
if so, to recall the age (in
years and months) it
occurred.

Age at outcome:
age of menarche

age, maternal parity,
maternal education,
and prenatal calcium
treatment status; Cox
regression models,
using self-reported age
at menarche as the
time to event, were
adjusted for maternal
age, maternal parity,
maternal education,
and prenatal calcium
treatment status; Cox
regression models were
also restricted to girls
<14.5 yr at the time of
the interview and
adjusted for maternal
age, maternal parity,
maternal education,
and prenatal calcium
treatment status

Second trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.59 (0.28, 0.90)
Third trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.85 (0.42, 1.27)

Cox models

First trimester maternal

blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.92 (0.65, 1.29)
Second trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.91 (0.65, 1.27)
Third trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.97 (0.69, 1.37)

Cox models restricted to
girls <14.5 yr at interview
First trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.80 (0.52, 1.25)
Second trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.64 (0.38, 1.09)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Third trimester maternal
blood

<5 [jg/dL: Reference
>5 [jg/dL: 0.89 (0.56, 1.41)

Effects on Puberty Among Males

Yao et al. (2019)
United States

2011-2012

Cross-sectional

NHANES

n: 431 male children,
493 male adolescents

Male children (age 6-
11 yr) and male
adolescents (age 12-
19 yr) in NHANES
2011-2012

Blood

Blood was measure by ICP-
MS

Age at Measurement:
6-19 yr old

Geometric mean
Male children: 0.76 [jg/dL
Male adolescents: 0.68 [jg/dL
Median

Male children: 0.72 [jg/dL
Male adolescent: 0.66 [jg/dL
75th

Male children: 1.02 [jg/dL
Male adolescents: 0.96 [jg/dL

Quartiles (|jg/dL):

Male children:

Q1
Q2
Q3
Q4

<0.52
0.52-0.72
0.72-1.02
>1.02

Male adolescents:
Q1: <0.47

Puberty among males:
Serum tT levels in male
children and adolescents

Serum tT levels were
analyzed by isotope-
dilution liquid
chromatography-tandem
mass spectrometry

Age at outcome:
6-19 yr old

Weighted multivariable
linear regression
models; Model 1
controlled forage, race,
and BMI. Model 2
controlled for PIR,
seasons of collection,
times of venipuncture,
and serum cotinine, in
addition to the
covariates of model 1

(3 (95% CI), as percent
difference in serum tT

Model 1:

Male children

Q1: Reference

Q2: 4.1 (-18.47, 32.9)

Q3: -6.13 (-27.64, 21.77)

Q4: -12.83 (-33.68, 14.58)

p for trend: 0.36

Male adolescents

Q1
Q2
Q3
Q4

Reference
-3.36 (-20.98, 18.2)
14.99 (-7.77, 43.37)
15.62 (-7.07, 43.86)

p for trend: 0.18

Model 2:

Male children

Q1: Reference

Q2: 11.75 (-13.06, 43.65)

Q3: -4.63 (-26.97, 24.55)

Q4: -13.09 (-34.45, 15.22)

p for trend: 0.42

Male adolescents

Q1: Reference

Q2: -4.35 (-21.22, 16.14)

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa



Q2: 0.47-0.66
Q3: 0.66-0.96
Q4: >0.96





Q3: 8.15 (-12.91, 34.3)
Q4: 6.32 (-14.62, 32.4)
p for trend: 0.58

De Craemer et al.

(2017)

Belgium

FLEHS I: 2002-2006,
FLEHS II: 2007-
2011, and FLEHS III:
2012-2015

Cross-sectional

FLEHS I, FLEHS II
and FLEHS III
FLEHS I n: 1659,
FLEHS II n: 606, and
FLEHS III n: 406

Adolescents aged 14-
15 yr

Blood

Blood was analyzed by ICP-
MS

Age at Measurement:
14-15 yr old

Geometric meanb
FLEHS I: 2.13 pg/dL
FLEHS II: 1.38 pg/dL
FLEHS III: 0.926 pg/dL

Maxb:

FLEHS I: 21.2 pg/dL
FLEHS II: 7.69 pg/dL
FLEHS III: 3.86 pg/dL

Puberty among males:
Hormones and sexual
maturation in adolescents

Development of genitals
in adolescent males and
pubic hair was scored
using the international
scoring criteria of
Marshall and Tanner,
where stage 1
corresponds to the start of
puberty and stage 5 to the
adult stage. Sex
hormones investigated in
this study were E2,
testosterone, fE2 and fT,
SHBG, LH, and FSH.
Hormone levels in
adolescent males were
measured in blood serum
using commercial
immunoassays.

Age at outcome:

14-15 yr old

Logistic regression
models for male public
hair development and
genital development
were adjusted for age
and BMI; linear
regression models for
hormones (ratio T/E2,
E2, fE2, T, IT) were
adjusted for age, hr of
blood collection, BMI,
smoking status; SHBG:
age, fasting, BMI,
smoking status, hr of
blood collection; LH
and FSH: age, BMI,
smoking status

OR (95% Cl)c
Pubic hair development
FLEHS I: 0.808 (0.686,
0.949)

FLEHS II: 0.849 (0.563,
1.365)

FLEHS III: 0.515 (0.327,
0.774)

Genital development

FLEHS I: 0.843 (0.717,

0.99)

FLEHS II: 0.697 (0.462,
0.998)

FLEHS III: 0.621 (0.388,
0.967)

(3 (95% Cl)c
FLEHS I:

Ratio T/E2: 1.022 (0.985,
1.059)

E2: 1.011 (0.991, 1.031)
fE2: 1.003 (0.975, 1.033)
T: 1.039 (0.993, 1.087)
fT: 1.026 (0.967, 1.09)
SHBG: 1.024 (0.992, 1.056)
LH: 0.995 (0.959, 1.033)
FLEHS II:

ratio T/E2: 1.002 (0.958,
1.049)

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Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

E2: 0.968 (0.923, 1.016)
fE2: 0.908 (0.839, 0.983)
T: 0.959 (0.906, 1.015)
IT: 0.909 (0.828, 0.997)
SHBG: 1.005 (0.961, 1.052)
LH: 0.974 (0.923, 1.028)
FSH: 0.995 (0.942, 1.05)

Nkomo et al. (2018)

Johannesburg
South Africa

Cohort

BT20+ birth cohort
n: 683

Singleton births in
which the infant
resides in

Johannesburg area for
at least 6 mo after
birth; participants must
have data for BLL at
age 13 and pubertal
growth trajectory
classes

Blood and cord blood

UCB collected at birth and
blood at collected at age 13
were measured by AAS with
a Zeeman background
correction

Age at Measurement:
birth and age 13

Mean (SD)

UCB: 5.9 (2.0) pg/dL

Blood: 6.6 (2.6) pg/dL

Median

UCB: 6.0 pg/dL
Blood: 6.5 pg/dL

75th

UCB: 7.0 pg/dL
Blood: 6.0 pg/dL

Puberty among males:
Pubertal trajectory
classes

Tanner stages of pubertal
development refer to a
standard clinical method
used to describe physical
measurements of
secondary sexual
characteristics using
drawings to signal stage
of pubertal development
where stage 1 signifies
lowest level of pubertal
maturation and stage 5
denotes highest level of
pubertal maturation in
boys

Age at outcome:
9-16 yr old

Multinomial logistic
regression models were
used to predict pubertal
growth trajectory class
based on (1) UCB Pb
and adjusted for
ethnicity; (2) blood Pb
and adjusted for
ethnicity and height at
age 8

RR (95% CI)

UCB

Pubic hair development
Trajectory Class 1:
Reference

Trajectory Class 2: 0.61
(0.25, 1.43)

Trajectory Class 3: 0.28
(0.11, 0.74)

Genital development

Trajectory Class 1:
Reference

Trajectory Class 2: 0.27
(0.03, 2.26)

Trajectory Class 3: 0.24
(0.03, 1.89)

Trajectory Class 4: 0.13
(0.01, 1.24)

Blood

Pubic hair development

Trajectory Class 1:
Reference

Trajectory Class 2: 0.94
(0.63, 1.39)

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Study Design

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Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Trajectory Class 3: 1.35
(0.73, 2.47)

Genital development

Trajectory Class 1:
Reference

Trajectory Class 2: 0.77
(0.33, 1.77)

Trajectory Class 3: 0.88
(0.38, 2.01)

Trajectory Class 4: 1.02
(0.37, 2.83)

Liu et al. (2019b)

Mexico City
Mexico

Cohort

n: 547 (283 girls and
264 boys)

Pregnant women were
recruited at three
public maternity
hospitals (Manuel Gea
Gonzalez Hospital,
Mexican Social
Security Institute and
the National Institute
of Perinatology) in
Mexico City; and
Children at age 9.8-
18.0 yr who had at
least one
measurement of
maternal bone Pb or
childhood blood Pb

Blood and bone

Maternal bone was measured
at the mid-tibial shaft (cortical
bone) and patella (trabecular
bone) and determined using
the X-ray fluorescence
instrument; blood samples
from children were measured
by GFAAS

Age at Measurement:
Maternal age 1-mo
postpartum; blood measured
between 1 and 4 yr

Median

Patella: 7.44 |jg/g
Tibia: 7.10 |jg/g
Blood, cumulative 1-4 yr:
14.33 [jg/dL

75th

Patella: 14.56 |jg/g

Puberty among males:
Pubertal stages

In boys, the stage of
sexual maturation was
defined by the
pediatrician using Tanner
staging scales for the
development of genitalia
and pubic hair.

Age at outcome:
9.8-18 yr

Ordinal regression
models for genitalia
and pubic hair and
logistic regression
models for TV were
adjusted for adjusted
for child age at visit,
maternal education and
marital status, and
number of siblings at
birth

OR (95%), per IQR increase
in Pb

Genital development

Patella: 0.963 (0.734,

1.264)

Tibia: 1.00 (0.711, 1.406)
Blood, cumulative 1-4 yr:
0.995 (0.948, 1.044)

Pubic hair development
Patella: 1.094 (0.836,

1.432)

Tibia: 1.00 (0.715, 1.398)

Blood, cumulative 1-4 yr:
1.004 (0.969, 1.04)

TV

Patella: 1.158 (0.804,

1.667)

Tibia: 0.885 (0.503, 1.558)
Blood, cumulative 1-4 yr:
1.013 (0.954, 1.075)

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Outcome

Confounders

Effect Estimates and 95%
Clsa

Reference and
Study Design

Study Population

Exposure Assessment

Tibia: 15.93 |jg/g

Blood, cumulative 1-4 yr:
18.90 [jg/dL

IQR

Patella: 13.57 |jg/g

Tibia: 13.30 |jg/g

Blood, cumulative 1-4 yr:
7.66 [jg/dL

Williams et al

Chapaevsk
Russian

2003-2005(2017)
Cohort

Russian Children's

Study

n: 516

Healthy male children
who were 8-9 yr old
between 2003 and
2005 in Chapaevsk,
Russia.

Blood

Blood was measured by
Zeeman background
corrected flameless GFAAS

Age at Measurement:
8-9 yr old

Median: 3 [jg/dL
Max: 31 [jg/dL

Puberty among males:
Male sexual maturity

Pubertal status was
staged from 1 to 5 via
examination by a single
clinician according to
internationally accepted
criteria. Pubarche (pubic
hair stage, P) was
determined by the extent
of terminal hair growth.
Genital staging (G) was
assessed by genital size
and maturity. TV was
measured using an
orchidometer. Three
different measures of
sexual maturity were
considered as separate
indicators: TV >20 mL of
either testis, genitalia
stage 5 (G5), and pubic
hair stage 5 (P5).
Duration of pubertal
progression was defined
as time from pubertal
onset (TV >3 mL,
genitalia stage >2 (G2),

Interval-censored
models were fit
assuming a normal
distribution for age at
sexual maturity using
accelerated failure time
models to compare
pubertal outcomes
between boys with
'higher' (>5 |jg/dL)
versus 'lower'

(<5 |jg/dL) peripubertal
BLLs. Models were
adjusted for boy's BW,
prenatal exposure to
maternal alcohol and
tobacco, maternal age
at son's birth,
household

characteristics including
income level, parental
education, and whether
the biological father
lived in the same
household, the boy's
physical activity, and
his nutritional status
determined by caloric

(3 (95% Cl)c, as shift in
mean age in months
Age at pubertal onset

Genitalia (G2): 8.40 (3.70,
13.10)

Pubic hair (P2): 8.12 (3.46,
12.78)

TV (>3 mL): 7.68 (3.46,
11.90)

Age at sexual maturity

Genitalia (G5): 4.20 (0.56,
7.84)

Pubic hair (P5): 4.23 -0.31,
8.77)

TV (>20 mL): 5.14 (1.70,
8.58)

Duration of puberal
progression

Genitalia (G2 to G5): -3.76
(-7.93, 0.42)

Pubic hair (P2 to P5): -1.82
(-6.91, 3.28)

TV (>3 mL to >20 mL):
-1.19 (-4.92, 2.54)

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Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

pubic hair stage >2 (P2),
respectively) to sexual
maturity, separately for
each pubertal indicator.

Age at outcome:
age at follow-up in 2017

intake and percent of
fat and protein intake.

Mediation analysis was
conducted to partition
the effect of higher vs.
lower BLLs on the age
at sexual maturity into a
direct effect of Pb
exposure and indirect
effect of Pb acting
through HTZ and BMIZ
(mediators) at age 11.

Mediation Analysis, as % of
total

HTZ

G5: 53.0% ((3: 2.37 mo)

P5: 47.5% ((3: 2.36 mo)

TV >20 mL: 34.2% ((3:
1.78 mo)

BMIZ

G5: 14.3% ((3: 0.64 mo)

P5: 23.4% ((3: 1.16 mo)

TV >20 mL: 6.1% ((3:
0.32 mo)

Fleisch et al. (2013)

Chapaevsk
Russia

2003-2005
Follow-up: 2-yr (at
10-11 yr) and 4-yr (at
12-13 yr)

Cohort

Russian Children's

Study

n: 394

Boys ages 8-9 yr old
from Chapaevsk,
Russia

Blood

Blood was measured by
Zeeman background
corrected flameless GFAAS

Age at Measurement:
8-9 yr old

Median: 3 [jg/dL
75th: 5 [jg/dL
Max: 31 [jg/dL

Puberty among males:
IGF-1

Serum IGF-1
concentrations were
measured by a
chemiluminescent
immunometric assay
using Siemens Immulite
2000.

Age at outcome:

10-11 yr (at 2-yr follow-
up); 12-13 (at 4-yr follow-
up)

Linear regression
models using a GEE
approach to account for
the repeated measures
were fitted to predict
the mean levels of
serum concentrations
of IGF-1 (ng/mL) in
relation to BLLs,
adjusted for baseline
parental education,
BW, nutritional intake,
and baseline and
follow-up age and BMI

(3 (95% CI)c, as adjusted

mean change

BLL <5 [jg/dL: Reference

BLL >5 ug/dL: -29.2 ng/mL
(-43.8, -14.5)

Pre-puberty

BLL <5 [jg/dL: Reference

BLL >5 ug/dL: -14.1 ng/mL
(-0.9, -27.2)

Early puberty:

BLL <5 [jg/dL: Reference

BLL >5 [jg/dL: -18.0 (-3.5,
-32.5)

Mid-puberty

BLL <5 [jg/dL: Reference

BLL >5 ug/dL: -41.9 ng/mL
(-15.1, -68.7)

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Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

AAS = atomic absorption spectrometry; BMI = body mass index; BMIZ = BMI-for-age Z-score; BT20+ = Birth to Twenty Plus; BW = birth weight; E2 = estradiol; ELEMENT = Early
Life Exposure in Mexico to Environmental Toxicants; fE2 = free estradiol; FLEHS = Flemish Environment and Health Study; FSH = follicle stimulating hormone; fT = free
testosterone; GEE = generalized estimating equation; GFAAS = graphite furnace atomic absorption spectrometry; HR = hazard ratio; HTZ = height Z-score; ICP-MS = inductively
coupled plasma mass spectrometry; IGF-1 = insulin-like growth factor 1; LH = luteinizing hormone; mo = month(s); NHANES = National Health and Nutrition Examination Survey;
OR = odds ratio; PIR = poverty-income ratio; RR = relative risk; SD = standard deviation; SHBG = sex hormone binding globulin; T = testosterone; tT = total testosterone;
TV = testicular volume; UCB = umbilical cord blood; yr = year(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect
estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated
interval. Categorical effect estimates are not standardized.
bPb measurements were converted from |jg/L to |jg/dL.

°Effect estimates unable to be standardized.

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Table 8-13 Epidemiologic studies of exposure to Pb and other developmental effects

study Desfgn Study P°Pulation ExP°sure Assessment	Outcome	Confounders	Effect	and 95%

Aleqrfa-Torres et al

Salamanca
Mexico

Cross-sectional

n: 86

Healthy children 6-
15 yr of age were
recruited from four
primary schools

Blood

Blood was measured by
ICP-MS

Age at Measurement:
6-15 yr old

Mean (SD): 3.78
(3.73) [jg/dL
Max: 22.61 pg/dL

Other developmental
effects: Telomeric
lengthening and mtDNA
effects

DNA was isolated from
peripheral blood and rTL
and the mtDNAcn were
determined by real-time
polymerase chain reaction

Age at outcome:
6-15 yr old

Linear regression
analyses; TL models
were adjusted for
mtDNAcn, sex, age, and
total white blood cell
count; mtDNAcn models
adjusted for TL, sex,
age, and total white
blood cell count

(3 (95% Cl)b

TL: 0.088 (-0.027, 0.097)

mtDNAcn: -0.198 (-2.81,
-0.17)

Tamayo v Ortiz et al.

Mexico City
Mexico

2007-2011

Cohort

PROGRESS birth
cohort

n: 255 for 12 mo
n: 150 for 18-24 mo

Women were invited to
participate during their
prenatal care visits at 4
clinics belonging to the
Mexican Social Security
System

Blood and bone

Maternal blood, collected
twice during pregnancy
(second and third
trimesters), was
measured by ICP-MS.
Maternal bone, from the
mid-tibial shaft, was
measured using a K-XRF
instrument during the first
month postpartum visit

Age at Measurement:
Maternal age at second
trimester, third trimester,
and 1 mo postpartum

Mean

2nd trimester blood for 12-
mo-old infants: 3.5 pg/dL

Other developmental
effects: Cortisol levels

Four saliva samples per
day from their child at
home; saliva samples
were analyzed in
duplicate using a
chemiluminescence-assay

Age at outcome:
12 or 18-24 mo

Longitudinal functional
mixed effects regression
models with penalized
splines were adjusted for
child's sex and maternal
age at delivery,
education, and pre-
pregnancy BMI

(3 (95% Cl)b

12-mo infants

Second trimester maternal
blood

Lower Pb: Reference
Moderate Pb: -0.07 (-0.24,
0.10)

Higher Pb: -0.51 (-0.85,
-0.18)

Third trimester maternal
blood

Lower Pb: Reference

Moderate Pb: -0.14 (-0.31,
0.03)

Higher Pb: -0.02 (-0.31,
0.26)

Tibia

Lower Pb: Reference

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

2nd trimester blood for
18-24-mo-old infants:
3.9 [jg/dL

3rd trimester blood for 12-
mo-old infants: 3.7 [jg/dL
3rd trimester blood for 18-
24-mo-old infants:
4.2 [jg/dL
Tibia for 12-mo-old
infants: 5.6 |jg/g

Tibia for 18-24-mo-old
infants: 4.9 |jg/g

Tertiles

Lower Pb: <5 [jg/dL
Moderate Pb: 5 < Pb
<10 |jg/dL

High Pb: >10 [jg/dL

Moderate Pb: 0.02 (-0.14,
0.19)

Higher Pb: -0.03 (-0.21,
0.14)

18-24-mo infants

Second trimester maternal
blood

Lower Pb: Reference
Moderate Pb: 0.11 (-0.08,
0.30)

Higher Pb: 0.23 (-0.19, 0.65)

Third trimester maternal
blood

Lower Pb: Reference
Moderate Pb: 0.01 (-0.17,
0.20)

Higher Pb: -0.05 (-0.51,
0.41)

Tibia

Lower Pb: Reference
Moderate Pb: 0.10 (-0.13,
0.32)

Higher Pb: 0.14 (-0.08, 0.35)

Hou et al. (2020)

Guiyu and Haojiang
China

November-
December 2017

Cross-sectional

n: 574 (357 from Guiyu
and 217 from Haojiang)

Children 2.5-6 yr of
age that lived in Guiyu,
an e-waste
contaminated town or
Haojiang, a city with

Blood

Blood was measured by
GFAAS

Age at Measurement:
2.5-6 yr old

Median

Other developmental
effects: Oral anti-
inflammatory potential

Participants were
instructed to sit up straight
and slightly forward in
their chair. A sputum cup
was used to collect the
saliva. Decayed

Multivariable linear
regression model
adjusted for gender, age,
BMI, outdoor activities,
the sucking/biting of toys
and pencils, diet (sweet
consumption, bean
products, marine
products), family
member smoking,

(3 (95% CI)b: -3.65 (-8.07,
0.77)

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

similar culture but no e-
waste recycling activity

Reference group:
3.47 [jg/dL

Exposed group:
4.86 [jg/dL
75th

Reference group:
4.07 [jg/dL
Exposed group:
4.86 [jg/dL

deciduous teeth were
detected under natural
and artificial light. The
concentration of salivary
sialic acids was
determined using a
quantitative competitive
ELISA kit.

Age at outcome:
2.5-6 yr old

paternal education
levels, monthly
household income

Sitarik et al. (2020)

Detroit, Ml
United States

September 2003-
December 2007
(December 2011-
September 2019)

Cohort

WHEALS birth cohort
n: 146

All women were in their
second trimester or
later, were aged 21-
49 yr, and were living in
a predefined
geographic area in
Wayne and Oakland
counties of Michigan.
Teeth were selected for
metal measurement if

(1)	the child had at
least some outcome
data available (birth
outcomes and/or a 2-yr
clinic visit) or early life
microbiome data; and

(2)	the tooth sample
met laboratory quality
control/quality
assurance guidelines

Teeth

Teeth were measured by
LA-ICP-MS. Teeth were
sectioned, and the
neonatal line (a
histological feature formed
in enamel and dentine at
the time of birth) and
incremental markings
were used to assign
temporal information to
sampling points. Second
trimester, third trimester,
postnatal (birth through 1
yr), and childhood (age 1
to tooth shedding) Pb
levels.

Age at measurement:
Estimated exposure from
2nd trimester, 3rd
trimester, and postnatally
(<1 yr of age)

Other developmental
effects: Gut microbiota (in
infants)

Families were asked to
retain the most recent
soiled diaper prior to the
home visit and stool
samples from infants ages
1-6 mo.

Age at outcome:

1-6 mo

Permutational
multivariate analysis of
variance models were
adjusted for tooth type,
tooth attrition, tooth
batch, exact age at stool
sample collection, and
child race

(3 (SE)b

Alpha diversity metrics
Second trimester
Richness - Bacterial
1 mo: 5.53 (6.98)
6 mo: -7.77 (7.31)
Richness - Fungal
1 mo: 0.29 (1.65)
6 mo: 1.7 (1.51)

Evenness - Bacterial
1 mo: 0 (0.01)

6 mo: -0.02 (-0.01)
Evenness - Fungal
1 mo: 0.03 (0.05)
6 mo: -0.02 (0.05)

Faith's Diversity - Bacterial
1 mo: 0.16 (0.39)
6 mo: -0.19 (0.37)

Faith's Diversity - Fungal
1 mo: Not reported
6 mo: Not reported

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Shannon Diversity -
Bacterial

1 mo: 0.01 (0.08)

6 mo: -0.11 (0.07)

Shannon Diversity - Fungal

1 mo: 0.06 (0.15)

6 mo: 0 (0.14)

Third trimester

Richness - Bacterial

1 mo: 2.52 (6.37)

6 mo: -13.11 (8.36)

Richness - Fungal

1 mo: 0.69 (1.82)

6 mo: 2.54 (1.56)

Evenness - Bacterial

1 mo: -0.01 (0.01)

6 mo: -0.02 (-0.01)

Evenness - Fungal

1 mo: 0.03 (0.05)

6 mo: 0.03 (0.05)

Faith's Diversity - Bacterial

1 mo: 0.03 (0.35)

6 mo: -0.52 (0.42)

Faith's Diversity - Fungal

1 mo: Not reported

6 mo: Not reported

Shannon Diversity -
Bacterial

1 mo: -0.05 (0.07)
6 mo: -0.12 (0.08)

Shannon Diversity - Fungal
1 mo: 0.09 (0.16)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

6 mo: 0.15 (0.15)

Postnatal

Richness - Bacterial

1 mo: 2.18 (7.16)

6 mo: -2.55 (6.42)

Richness - Fungal

1 mo: -1.85 (2.54)

6 mo: -0.35 (1.05)

Evenness - Bacterial

1 mo: -0.02 (0.01)

6 mo: -0.01 (0.01)

Evenness - Fungal

1 mo: 0.07 (0.1)

6 mo: 0.06 (0.06)

Faith's Diversity - Bacterial

1 mo: -0.08 (0.39)

6 mo: 0.11 (0.32)

Faith's Diversity - Fungal

1 mo: Not reported

6 mo: Not reported

Shannon Diversity -
Bacterial

1 mo: -0.1 (-0.08)
6 mo: -0.05 (-0.06)
Shannon Diversity - Fungal
1 mo: -0.07 (0.23)
6 mo: -0.05 (0.1)

BMI = body mass index; ELISA = enzyme-linked immunosorbent assay; GFAAS = graphite furnace atomic absorption spectrometry; ICP-MS = inductively coupled plasma mass
spectrometry; K-XRF = K-shell X-ray fluorescence instrument; LA-ICP-MS = laser ablation-inductively coupled plasma-mass spectrometry; mo = month(s); mtDNA = mitochondrial
DNA; mtDNAcn = mitochondrial DNA copy number; rTL = relative telomere length; SD = standard deviation; SE = standard error; TL = telomere length; WHEALS = Wayne County
Health, Environment, Allergy and Asthma Longitudinal Study; yr = year(s).

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study Desfgn Study P°Pulation ExP°sure Assessment	Outcome	Confounders	Effect Esti™£s and 95%

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect
estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated
interval. Categorical effect estimates are not standardized.
bEffect estimates unable to be standardized.

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Table 8-14

Epidemiologic studies of exposure to Pb and female reproductive effects



Reference and
Study Design

Study Population Exposure Assessment Outcome Confounders

Effect Estimates and 95%
Clsa

Effects on Hormones Levels and Menstrual/Estrous Cycle

Krieq and Feng
(2011)

United States

1999-2002

Cross-sectional

NHANES
n: 649

Women aged 35-60 yr
old

Blood

Blood was measured by
AAS

Age at Measurement:
35-60 yr

Geometric mean:
1.4 |jg/dL Mean:
1.6 Mg/dL
Max: 17.0 pg/dL

Female reproductive
function: Serum FSH and
LH

Serum FSH and LH were
measured using a
microparticle enzyme
immunoassay

Age at outcome:

35-60 yr

Regression analyses:
the slopes were
adjusted forage, Iog10
serum bone alkaline
phosphatase, log 10
urine N-telopeptides,
Iog10 serum cotinine,
alcohol use, currently
breastfeeding,
hysterectomy, one
ovary removed, Depo-
Provera use, medical
conditions or
treatments, hormone
pill use, and hormone
patch use

(3 (95% CI)b, as slope for
serum FSH and LH per
Iog10 blood Pb increase

Serum FSH (IU/L)
Post-menopausal: 26.38
(13.39, 39.38)

Pregnant: -0.08 (-1.11,
0.95)

Menstruating: 1.50 (-2.29,
5.30)

Both ovaries removed:
27.71 (1.64, 53.78)

Birth control pills: -0.33
(-6.52, 5.86)

Pre-menopausal: 11.97
(3.27, 20.66)

Serum LH (IU/L)
Post-menopausal: 11.63
(4.40, 18.86)

Pregnant: 2.12 (-14.62,
18.86)

Menstruating: 0.87 (-2.20,
3.94)

Both ovaries removed:
20.59 (2.14, 39.04)

Birth control pills: 2.19
(-1.35, 5.72)

Pre-menopausal: 7.44
(-0.26, 15.14)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Chen et al. (2016)

Shanghai, Jiangxi
Province and
Zhejiang Province
China

2014

Cross-sectional

SPECT-China
n: 2286 men and 1571
postmenopausal women

SPECT-China is a
population-based cross-
sectional survey on the
prevalence of metabolic
diseases and risk factors
in East China. Men and
postmenopausal women
(age >55 yr) who were
not taking hormone
replacement therapy,
without a history of
hysterectomy and
oophorectomy were
recruited.

Blood

Blood was measured by
AAS

Age at measurement:
Median age 63 (IQR: 59-
68)

Median0: 4.1 [jg/dL
75thc: 5.981 pg/dL

Quartile0 (pg/dL)

Q1
Q2
Q3
Q4

<2.7

2.7-4.099
4.1-5.980
>5.980

Female reproductive
function: Reproductive
hormone levels

Venous blood samples were
drawn from all subjects after
an overnight fast of at least
8 hr. HbA1c was assessed
via high-performance liquid
chromatography (MQ-
2000PT, China). tT, E2, LH
and FSH levels were
measured using
chemiluminescence assays
(Siemens Immulite 2000,
Germany). SHBG levels
were detected using Cobas
e601

electrochemiluminescence
immunoassays (Roche,
Switzerland).

Age at outcome:

Median age 63 (IQR: 59-68)

Linear regression
models were adjusted
for age, current
smoking status, BMI,
SBP, diabetes, and
blood Cd level

(3 (SE)d
SHBG

Q1
Q2
Q3
Q4

tT

Q1

Q2

Q3

Q4

E2
Q1
Q2
Q3
Q4

Reference
0.010 (0.015)
0.018 (0.015)
0.048 (0.016)

Reference
-0.033 (0.019)
-0.017 (0.019)
-0.016 (0.020)

Reference
-0.001 (0.019)
-0.020 (0.019)
-0.021 (0.020)

FSH

Q1

Q2

Q3

Q4

Reference
0.013 (0.015)
0.047 (0.015)
0.046 (0.016)

LH
Q1
Q2
Q3
Q4

Reference
0.022 (0.015)
0.027 (0.016)
0.037 (0.016)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Lee et al. (2019)

Busan
Korea

2012-2014

Cross-sectional

Second Korean National Blood

Environmental Health
Survey

n: 4,689 adults

2,763 men and 1,926
postmenopausal women
aged 50 yr or over

Blood was measured by
GFAAS

Age at Measurement:
50 yr or older

Median: 2.05 [jg/dL
75th: 2.67 pg/dL

Female reproductive
function: Follicle-stimulating
hormone levels

Serum FSH levels were
measured using a
chemiluminescence
immunoassay
(chemiluminescent
immunoassay; ADVIA
Centaur XP; Siemens,
Tarrytown, NY, United

Multiple linear
regression adjusted for
age, BMI, smoking
status, and alcohol
consumption

(3 (95% CI)b: 2.929 (0.480,
5.377)

tendola et al. (2013)

United States

1999-2010

Cross-sectional

NHANES
n: 3,221 (2,158
menstruating and 1,063
menopause)

Women aged 45-55 yr

Blood

Blood was measured by
AAS in 1999-2002 and
ICP-MS in 2003-2010

Age at measurement: 45-
55 yr

Geometric mean:

Menopausal women:
1.71 pg/dL

Menstruating women
1.23 pg/dL

Quartiles (pg/dL)

Q1
Q2
Q3
Q4

LOD-1.0

1.0-1.4
1.4-2.1

2.1-22.4

Female reproductive:
Menopause

Menopause was
dichotomized: women with
at least one menstrual cycle
in the past 12 mo were
categorized as "No" and
those with natural
menopause were "Yes"

Age at outcome:

45-55 yr

Logistic regression
models were adjusted
for age, race/ethnicity,
current hormone use,
poverty, and smoking;
NHANES 1999-2002
models also adjusted
for bone alkaline
phosphatase; and
NHANES 2005-2008
models also adjusted
for femoral neck bone
density

OR (95% CI)
NHANES 1999-2010

Q1
Q2
Q3
Q4

Reference
1.7 (1.0, 2.8)
2.1 (1.2, 3.6)
4.3 (2.6, 7.2)

NHANES 1999-2002

Q1: Reference

Q2: 1.0 (0.3, 3.5)

Q3: 1.3 (0.4, 4.5)

Q4: 5.1 (1.4, 18.0)

Adjusted for bone alkaline
phosphatase

Q1
Q2
Q3
Q4

Reference

1.1	(0.3, 3.9)

1.2	(0.3, 4.7)
4.2 (1.2, 15.5)

NHANES 2005-2008
Q1: Reference

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Q2: 3.0 (0.9, 9.8)
Q3: 4.9 (1.5, 16.1)
Q4: 10.5 (3.1, 35)
Adjusted for femoral neck
bone density

Q1
Q2
Q3
Q4

Reference
3.4 (0.9, 12.2)
4.1 (1.1, 15.2)
9.7 (2.8, 33)

Eum eta!. (2014)

Boston, MA
United States

1990-1994 (2001-
2004)

Cohort

Nurse's Health Study
n: 434

Female registered
nurses, 30 to 55 yr of age
and living in 11
U.S. states, completed a
questionnaire on their
medical history and
health-related behaviors;
analysis restricted to
women in the Boston
area who did not have a
history of a major,
chronic disease; and
were not obese from
1990-1994 and women
no history of chronic
diseases (no reported
diagnosis of
hypertension,
cardiovascular disease,
renal disease, diabetes,
or malignancies) invited
to participate from 2001
through 2004

Blood and bone

Bone was measured by
K-XRF at each woman's
mid-tibial shaft and
patella. Blood was
measured by GFAAS with
Zeeman background
correction

Age at measurement:
46 yr or older at the time
of bone Pb measurement

Median
Tibia: 10 |jg/g
Patella: 12 |jg/g
Blood: 3 [jg/dL
75th

Tibia: 15 |jg/g
Patella: 18 |jg/g
Blood: 4 [jg/dL

Tertiles
Tibia (|jg/g)

Female reproductive
function: Early menopause

Menopausal status was
determined on the first
Nurse's Health Study
questionnaire in 1976 and
then again on each biennial
questionnaire by asking
whether the participants'
menstrual periods had
ceased permanently; early
menopause as natural
menopause occurring
before 45 yr of age

Age at outcome:

Age at reporting of
menopausal status

Ordinary least-squares
linear regression to
analyze age at
menopause adjusted
for sub-study group,
age at bone Pb
measure, age at bone
Pb measure squared,
year of birth, age at
menarche, months of
oral contraceptive use,
parity, and pack-years
of smoking; logistic
regression for early
menopause adjusted
for sub-study group,
age at bone Pb
measure, age at bone
Pb measure squared,
year of birth, age at
menarche, months of
oral contraceptive use,
parity, and pack-years
of smoking

(3 (95% CI), as difference in
age at natural menopause
(year)

Tibia

T1
T2
T3

Reference
-0.80 (-1.67, 0.06)
-1.21 (-2.08, -.035)

p for trend: 0.006

Patella

T1
T2
T3

Reference
-0.32 (-1.18, 0.55)
-0.00 (-0.88, 0.87)

p for trend: 0.99

Blood

T1
T2
T3

Reference
0.08 (-0.80, 0.96)
-0.28 (-1.13, 0.56)

p for trend: 0.54

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

T1

<6.5

T2

6.513

T3

>13

Patella (pg/g)

T1

<8

T2

8-15

T3

>15

Blood (|jg/dL)

T1

<3

T2

3

T3

>3

Effects on Female Fertility

Lee et al. (2020)
United States

2013-2014 and
2015-2016

Cross-sectional

NHANES (2013-2014
and 2015-2016)
n: 124

Women aged 20-39 yr
without a history of
hysterectomy and/or
bilateral oophorectomy

Blood

Blood was measured by
ICP-MS

Age at Measurement:
20-39 yr

Geometric mean:

0.50 [jg/dL (95% CI: 0.43,

0.57)

Female reproductive
function: Female infertility

Infertility is defined as the
absence of pregnancy with
unprotected intercourse for
1 yr and was assessed
through a self-reported
questionnaire

Age at outcome:

20-39 yr

Logistic regression
analyses were adjusted
for age, ethnicity,
annual family income,
education, marital
status, smoking history,
alcohol consumption,
physical activity, and
BMI

OR (95% CI)b: 2.60 (1.05,
6.41) per 2-fold increase in
BLLs

OR (95% CI)

T1
T2
T3

Reference
5.40 (1.47, 19.78)
5.62 (1.13, 27.90)

Tertiles (pg/dL)

T1
T2
T3

0.11-0.38
0.41-0.62
0.63-5.37

Louis et al, (2012) LIFE Study

Blood

Female reproductive

Cox models for

OR (95% CI), as

n: 501



function: Fecundity

discrete survival time,

fecundability OR

Michigan (4 counties)

and Texas (12 Female ages 18-44 yr

Blood was measured by
ICP-MS

Women were instructed in

which is a proportional
odds model, adjusted

Female only exposure:
0.97 (0.85, 1.11)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

counties)
United States

2005-2009

Cohort

and male ages >18 yr; in
a committed relationship;
ability to communicate in
English or Spanish;
menstrual cycles
between 21 and 42 d; no
hormonal contraception
injections during past
year; and no sterilization
procedures or physician
diagnosed infertility

Age at Measurement:
19-40 yr

Geometric mean
Pregnant female:
0.66 [jg/dL
Not pregnant female:
0.76 [jg/dL

Tertiles (pg/dL)
T1: 0.23-0.57

T2
T3

0.58-0.78
0.79-5.84

the use of the Clearblue
Easy fertility monitors
consistent with the
manufacturer's guidance
commencing on day six for
tracking daily levels of E3G
and LH. Women also used
the digital Clearblue Easy
home pregnancy test upon
enrollment to ensure the
absence of pregnancy at
study start and on the day
menses was expected for
each cycle under
observation in the study.

Age at outcome:

Average age with
pregnancy: 29.8
Average age without
pregnancy: 30.6

for age, BMI, cotinine,
parity, serum lipids,
and site

(Texas/Michigan)

Couple exposure:

Female exposure: 1.06
(0.91, 1.24)

Male exposure: 0.82 (0.68,
0.97)

Lai etal. (2017)

Taipei
Taiwan

2008-2010

Cross-sectional

n: 190 infertile women
including 68 patients with
endometriosis and 122
controls

Women who visited the
infertility clinic first time
for a specific
gynecologist at Taipei
Medical University
Hospital; women with
diagnoses such as
ovarian cyst, premature
ovarian failure, repeated
implantation failure or
pregnancy were
excluded

Blood

Blood was measured by
ICP-MS

Age at measurement:

Mean age for women with
endometriosis: 35.3 (SD:
4.1)

Mean age for women
without endometriosis:
35.3 (SD: 5.0)

Geometric mean0

Female reproductive
function: Endometriosis
among infertile women

Endometriosis status was
determined by laparoscopy

Age at outcome:

Mean age for women with
endometriosis: 35.3 (SD:
4.1)

Mean age for women
without endometriosis: 35.3
(SD: 5.0)

Multivariate logistic
regression adjusted for
age, body fat
proportion, educational
level, age at menarche,
and regularity of
menstrual cycle

OR (95% CI)
T1: Reference
T2: 1.73 (0.77,
T3: 2.59 (1.11,

3.88)
6.06)

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Outcome

Confounders

Effect Estimates and 95%
Clsa

Reference and
Study Design

Study Population

Exposure Assessment

Women with
endometriosis:
1.337 [jg/dL
Women without
endometriosis:
0.853 [jg/dL

Median0
Women with
endometriosis:
2.130 [jg/dL
Women without
endometriosis:
0.464 [jg/dL

Tertiles0 (pg/dL)
T1: <0.38
T2: 0.38-3.05
T3: >3.05

Li et al. (2022)

Hefei
China

October 2019 -
January 2020

Cohort

n: 1184

Participants selected
from First Affiliated
Hospital of Anhui Medical
University while seeking
IVF treatment and
diagnosed infertility with
their partner. Inclusion
criteria: women were
aged between 20 and
45 yr; couples were
diagnosed with infertility
(failure to establish a
clinical pregnancy with
unprotected intercourse
for at least 1 yr); and IVF
indicators were tubal

Blood

Maternal blood (serum)
was measured by ICP-MS

Age at measurement:
Maternal age at collection
(day before oocytes were
retrieved for IVF); female
partner mean age was
30.22 yr

Geometric meane:
0.0877 [jg/dL

Mediane: 0.0924 [jg/dL

75the: 0.14399 pg/dL

Female reproductive
function - Effects on female
fertility: Fertility - successful
implantation, clinical
pregnancy

A serum hCG level
>25 mlU/mL on the 14th d
after embryo transfer was
considered as successful
implantation. Clinical
pregnancy was defined as
an ultrasound-confirmed
intrauterine pregnancy on
the 30th d after embryo
transfer.

Age at outcome:

Logistic regression
model for successful
implantation adjusted
for maternal age, BMI,
treatment protocol,
FSH levels, sperm
viability, cycle type,
and embryo quality.
Logistic regression
model for clinical
pregnancy adjusted for
maternal age, BMI,
treatment protocol,
endometrial thickness
on hCG day, and
embryo quality. Linear
regression models for
Mil rate, fertility rate,

OR (95%CI)b:

Successful implantation
Continuous: 0.85 (0.77,
0.94)

Tertiles

Low: Reference
Medium: 1.11 (0.75, 1.63)
High: 0.58 (0.40, 0.85)

Clinical pregnancy

Continuous: 0.95 (0.91,
0.99)

Tertiles

Low: Reference
Medium: 0.72 (0.37, 1.38)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

factor, ovulation failure,
or other factors for
female partner or male
factor or unexplained
fertility.

Tertilese (pg/dL)
Low: 0.002-0.065
Medium: 0.065-0.125
High: 0.125-0.481

Female partner mean age:
30.22 yr

2PN rate, blastocyte
rate, and high-quality
embryo rate were
adjusted for maternal
age, BMI, education
level, infertility type,
FSH and sperm
concentration

High: 0.56 (0.29, 1.06)

(3 (95% CI)b:

Mil rate: 0.090 (-0.024,

0.204)

Fertility rate: -0.033
(-0.151, 0.086)

2PN rate: -0.019 (-0.100,
0.062)

Blastocyst rate: 0.046
(-0.052, 0.144)

High quality embryo rate:
-0.143 (-0.322, -0.037)

Zhou et al. (2021a) n: 195

China

2018-2019

Cohort

Couples undergoing IVF.
Women with
endometriosis,
hydrosalpinx, abnormal
uterine cavity and men
with azoospermia, severe
oligozoospermia,
asthenospermia and
dysspermia were
excluded from the study.

Blood

Maternal blood (serum),
follicular fluid, and
seminal plasma from male
partner

Age at Measurement:
Female partner mean
age: 30.27 yr
Male partner mean age:
31.57 yr

Mean0

Maternal serum:
0.301 [jg/dL
Follicular fluid:

0.742 [jg/dL
Seminal plasma:
0.882 [jg/dL

Median0

Female reproductive
function - Effects on female
fertility: IVF outcome

The IVF outcomes included
were normal fertilization,
good embryo, blastocyst
formation, high-quality
blastocyst, pregnancy, and
live birth

Age at outcome:

Female partner mean age:
30.27 yr

Male partner mean age:
31.57 yr

Poisson regression
models were adjusted
for age and BMI

RR (95% Cl)b
Normal fertilization
Maternal serum: 0.94
(0.42, 1.93)

Follicular fluid: 0.82 (0.18,
2.39)

Seminal plasma: 1.55
(0.64, 3.3)

Good embryo

Maternal serum: 1.00
(0.36, 2.38)

Follicular fluid: 0.78 (0.09,
3.03)

Seminal plasma: 1.86
(1.05, 3.11)

Blastocyst formation

Maternal serum: 1.06 (0.2,
3.91)

Follicular fluid: 0.41 (0,
3.63)

8-217


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Outcome

Confounders

Effect Estimates and 95%
Clsa

Seminal plasma: 1.77
(0.78, 3.58)

High-quality blastocyst
Maternal serum: 1.68
(0.15, 9.43)

Follicular fluid: 0.35 (0,
7.11)

Seminal plasma: 2.66
(0.67, 8)

Pregnancy
Maternal serum: 0.18
(0.01, 1.91)

Follicular fluid: 0.01 (0,
0.03)

Seminal plasma: 0.04 (0,
1.45)

Reference and
Study Design

Study Population

Exposure Assessment

Maternal serum:
0.245 [jg/dL

Follicular fluid:
0.178 [jg/dL
Seminal plasma:
0.486 [jg/dL

75thc

Maternal serum:
0.317 [jg/dL
Follicular fluid:
0.326 [jg/dL
Seminal plasma:
1.245 [jg/dL

Live birth

Maternal serum: 0.25
(0.01, 2.8)

Follicular fluid: 0 (0, 0.09)

Seminal plasma: 0.01 (0,
1.08)

Effects on Morphology or Histology of Female Sex Organs (Ovaries, Uterus, Fallopian Tubes/Oviducts, Cervix, Vagina, and/or Mammary Glands)

Ye etal. (2017)
Seoul

South Korea

September to
November 2014

n: 288 (46 with fibroids
and 242 without)

Premenopausal women
between 30 and 49 yr
old, who were not
pregnant or
breastfeeding, whose
heavy metal levels at the

Blood

Blood was measured by
GFAAS

Age at Measurement:
30-49 yr

Female reproductive
function - Effects on
morphology and histology of
female sex organs: Uterine
fibroids

Diagnosis of uterine fibroids
was based on pelvic
ultrasonography and two

Logistic regression
models adjusted for
age, BMI, gravidity,
oral contraceptive pill
administration history,
regularity of menstrual
cycle, hemoglobin
level, and serum
cotinine levels; linear

OR (95% CI)b

Presence of uterine
fibroids: 1.39 (0.75, 2.56)

(3 (95% Cl)b

Volume of uterine fibroids:
0.12 (-2.26, 2.51)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Cross-sectional

time might have been
influenced by these
circumstances and might
have been less
representative of heavy
metal levels at the time of
diagnosis, and who had
received hysterectomies

Geometric mean:
1.36 [jg/dL

Quartiles (pg/dL)
Q1: <1.1
Q2: 1.1-1.3
Q3: 1.3-1.8
Q4: 1.8-3.2

questions

Age at outcome:
30-49 yr

regression models
were adjusted for age,
BMI, gravidity, oral
contraceptive pill
administration history,
regularity of menstrual
cycle, hemoglobin
level, and serum
cotinine levels

Q1
Q2
Q3
Q4

Reference
-0.42 (-2.69, 1.85)
0.85 (-1.67, 3.37)
-1.23 (-3.74, 1.29)

2PN = oocytes with two pronuclei; AAS = atomic absorption spectrometry; BMI = body mass index; d = day(s); E2 = estradiol; E3G = estrone-3-glucuronide; FSH = follicle stimulating
hormone; GFAAS = graphite furnace atomic absorption spectrometry; hCG = human chorionic gonadotropin; ICP-MS = inductively coupled plasma mass spectrometry;
IQR = interquartile range; IVF = in vitro fertilization; K-XRF = K-shell X-ray fluorescence instrument; LH = luteinizing hormone; LIFE = Longitudinal Investigation of Fertility and the
Environment; LOD = limit of detection; Mil = metaphase II; mo = month(s); NHANES = National Health and Nutrition Examination Survey; OR = odds ratio; SBP = systolic blood
pressure; SD = standard deviation; SHBG = sex hormone binding globulin; SPECT = Survey on the Prevalence in East China for Metabolic Diseases and Risk Factors; tT = total
testosterone; yr = year(s).

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect

estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated

interval. Categorical effect estimates are not standardized.

bEffects estimates unable to be standardized.

°Pb measurements were converted from |jg/L to |jg/dL.

dNo CIs provided.

ePb measurements were converted from ng/Lto |jg/dL.

8-219


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Table 8-15

Animal toxicological studies of Pb exposure and female reproductive effects

Study

Species (Stock/Strain), n, Sex Timing of Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Cory-Slechta et al.

(2013)

Mouse (C57BL/6)

Control (untreated), F, n = 16

100 ppm, F, n = 16

GD -61 to PND 365

Dams were dosed starting 2 mo
prior to mating. Offspring were
continued on the same
exposure as their dams until the
end of the experiment at 12 mo
of age.

0.22 [jg/dL for control
dams at weaning

12.12 [jg/dL for
100 ppm dams at
weaning

Litter Size,
Maternal
Body Weight

Weston et al.

Rat (Long-Evans)

Dams

Control (untreated), F, n = 20
50 ppm Pb, F, n = 19
Pups

Control (untreated), M/F, n = 12.4
(7/5.4 average number of male and
female pups per litter in control)

50 ppm Pb, M/F, n = 7.4 (6.3/1.1
average number of male and
female pups per litter in Pb NS
group)

GD -76 to PND 21

Dams were dosed via drinking
water starting 2-3 mo prior to
breeding. Exposure ended at
weaning (PND 21).

Dams (PND 21):

0.500 [jg/dL for control

7.72 [jg/dL for 50 ppm
Pb

Pups (PND 5-6):

0.603 [jg/dL for control
males

0.690 [jg/dL for control
females

15.7 [jg/dL for 50 ppm
Pb males

14.6 [jg/dL for 50 ppm
Pb females

Litter Size,
Number of
Litters

8-220


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Study

Species (Stock/Strain), n, Sex Timing of Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Betharia and Maher

Rat (Sprague-Dawley)

Control (untreated), F, n = 6 dams

10 |jg/mL Pb, F, n = 6 dams

GD Oto PND 20

Dams were dosed via drinking
water throughout pregnancy
until weaning (PND 20).

Pups:

PND 2

0.188 [jg/dL for control
9.03 [jg/dL for 10 |jg/ml_
Pb

Litter Size

PND 25:

0.088 [jg/dL for control
0.976 [jg/dL for
10 |jg/ml_ Pb

PND 60:

0.0244 [jg/dL for control

0.0318 [jg/dL for
10 |jg/ml_ Pb

Schneider etal. (2016) Mouse (C57BL/6)

Control (untreated), F, n = NR

100 ppm Pb, F, n = NR

GD -61 to PND 21

Dams were dosed via drinking
water starting 2 mo prior to
mating through lactation
(weaning assumed to be
PND 21).

Dams were also treated to a
non-stress or prenatal stress
condition. Only data from dams
in the non-stress condition were
used.

Dams at weaning
(assumed PND 21):
0.22 [jg/dL for control
12.61 [jg/dL for
100 ppm Pb

Pups (PND 5-6):
0.37 [jg/dL for control

10.2 |jg/dL for 100 ppm
Pb

Maternal
Body
Weight,
Litter Size

Saleh etal. (2018)

Rat (Sprague-Dawley)

Control (vehicle), F, n = 8

160 ppm Pb, F, n = 8
320 ppm Pb, F, n = 8

GD 1 to 20

Dams were dosed via oral
gavage. Authors report a
significant decrease in brain
weight occurred, indicating
potential overt toxicity.

Dams (GD 20):
5.1 [jg/dL for control
27.7 |jg/dL for 160 ppm
Pb

41.5 |jg/dL for 320 ppm
Pb

Maternal
Body Weight

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Study

Species (Stock/Strain), n, Sex

Timing of Exposure

Exposure Details
(Concentration, Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Baranowska-Bosiacka

Rat (Wistar)

GD 1 to PND 21

Dams were exposed via

NR for Dams

Sex Ratio

etal. (2013)

Control (untreated), F, n = 3 dams



drinking water throughout







0.1% Pb, F, n = 3 dams



pregnancy until weaning
(PND 21).

Pups (PND 28):





Control, M/F, n = 36 (17/19) pups



0.93 |jg/dL for control





0.1% Pb, M/F, n = 36 (18/18) pups





6.86 |jg/dL for 0.1% Pb



Saleh etal. (2019)

Rat (Sprague-Dawley)

Control (vehicle), F, n = 8 dams

160 ppm Pb, F, n = 8 dams
320 ppm Pb, F, n = 8 dams

GD 1 to 20

Dams were dosed via oral
gavage. Authors report a
significant decrease in brain
weight occurred, indicating
potential overt toxicity.

Dams (GD 20):
5.26 |jg/dL for control
23.9 |jg/dL for 160 ppm
Pb

42.9 |jg/dL for 320 ppm
Pb

Maternal
Body Weight

BLL = blood lead level; F = female; GD = gestational day; M = male; mo = month(s); NR = not reported; Pb = lead; PND = postnatal day; NS = non-stress.

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Table 8-16

Epidemiologic studies on exposure to Pb and male reproductive effects



Reference and
Study Design

Study Population Exposure Assessment Outcome Confounders

Effect Estimates and 95%
Clsa

Effects on Sperm/Semen Production, Quality, and Function

Li eta!. (2015)
Taiwan

May 2012 to
February 2013

Cross-sectional

n: 154

Male participants were
recruited from a
reproductive medical
center and did not have
obstructive azoospermia,
cryptorchidism,
varicoceles, hydrocele,
orchitis, or epididymitis;
did not have testicular
injury or underwent
testicular surgery before
the study period

Blood

Blood was measured by
ICP-MS

Age at Measurement:
Mean age: 34.8 yr

Mean (SD)b: 2.78 (1.85)
pg/dL

Male reproductive effects:
Seminal parameters

From semen samples the
following parameters were
assessed: sperm
concentration, semen
volume, number of sperm,
percentage of total motility
sperm, percentage of
progressive motility sperm,
and percentage of sperm
with normal morphology

Age at outcome:

Mean age: 34.8 yr

Multiple logistic
regression models were
adjusted for FSH, LH,
prolactin, and
testosterone were input
into the model and then
adjusted for age and
education

OR (95% CI)

Low-quality semen: 1.040
(1.011, 1.069)

Sperm concentration: 1.046
(1.015, 1.078)

Numbers of sperm: 1.041
(1.012, 1.071)

Total motility sperm: 1.057
(1.026, 1.089)

Progressive motility sperm:
1.047 (1.014, 1.080)

Sperm with normal
morphology: 1.071 (1.025,
1.118)

Sukhn et al. (2018)

Beirut
Lebanon

January 2003 and
December 2009

Cross-sectional

Environment and Male
Infertility study
n: 116

Male partners of infertile
heterosexual couples who
attended the fertility clinic
at the American University
of Beirut Medical Center
were recruited. Men were
18 to 55 yr of age, had a
BMI of 18 to 30 kg/m2, and
had not been on any
hormone therapy for the
past 6 mo, no diabetes,
endocrine disease,
fertility-related genetic
disorders, obstructive
azoospermia,

Blood and other: seminal
fluid

Blood and seminal fluid
were measured by ICP-
MS equipped with a cell
dynamic range

Age at Measurement:
18-55yr

Meanb
Blood

Overall: 3.121 pg/dL

Low-quality semen group:
5.198 [jg/dL

Male reproductive effects:
Semen quality

Participants with a semen
volume <1.5 mL, sperm
concentration
<15 million/mL, total count
<39 million, progressive
motility <32%, viability
<58%, and/or normal
WHO morphology <30%
were assigned to the low
quality semen group A.
Participants whose semen
analyses expressed better
results in all the above
parameters were assigned
to the normal-quality
semen group B. Sperm

Logistic regression;
age, cigarette smoking,
alcohol intake, and
period of sexual
abstinence

OR (95% CI)
Blood

Volume (<1.5 mL)

Q1
Q2
Q3
Q4

Reference
0.53 (0.11, 2.44)
0.24 (0.02, 2.24)
1.32 (0.33, 2.56)

p for trend: 0.26

Concentration (<15 M/mL)
Q1
Q2
Q3
Q4

Reference
0.51 (0.16, 1.63)
1.17 (0.37, 3.73)
1.58 (0.53, 4.68)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

cryptorchidism, varicocele, Normal quality semen

hydrocele, orchitis.
Epididymitis, and/or
history of testicular injury
or surgery

group 3.575 [jg/dL
Seminal fluid
Overall: 0.540 [jg/dL
Low-quality semen group:
1.626 [jg/dL

Normal quality semen
group: 1.285 |jg/dL

Medianb
Blood

Low-quality semen group:
3.257 [jg/dL

Normal quality semen
group: 3.098 [jg/dL

Seminal fluid

Low-quality semen group:
0.588 [jg/dL

Normal quality semen
group: 0.470 |jg/dL

Quartilesb (pg/dL)

concentration (million/mL)
and progressive motility
(%) were determined
manually using a Makler®
counting chamber. Total
sperm count (million) was
calculated as sperm
concentration * semen
volume. Sperm
morphology was
determined by high-power
magnification (* 1000) on
air-dried smears stained
with a Wright Giemsa stain
based on the WHO
guidelines.

Age at outcome:

18-55 yr

Q1
Q2
Q3
Q4

LOD-2.199
2.200-3.256
3.257-5.357
>5.358

p for trend: 0.26

Total count (<39 M)
Q1
Q2
Q3
Q4

Reference
0.36 (0.11, 1.18)
0.83 (0.26, 2.65)
1.35 (0.46, 3.96)

p for trend: 0.15

Progressive motility (<32%)
Q1
Q2
Q3
Q4

Reference
0.70 (0.19, 2.62)
0.78 (0.19, 3.19)
1.47 (0.43, 5.02)

p for trend: 0.66

Viability (<58%)

Q1
Q2
Q3
Q4

Reference
0.44 (0.14, 1.39)
0.68 (0.21, 2.21)
1.35 (0.46, 3.96)

p for trend: 0.23

WHO morphology (<30%)
Q1
Q2
Q3
Q4

Reference
0.50 (0.15, 1.66)
0.93 (0.28, 3.10)
0.84 (0.26, 2.66)

p for trend: 0.68

Seminal Fluid
Blood

Volume (<1.5 mL)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Q1
Q2
Q3
Q4

Reference
0.86 (0.16, 4.67)
1.34 (0.25, 7.17)
2.07 (0.37, 11.51)

p for trend: 0.95

Concentration (<15 M/mL)
Q1
Q2
Q3
Q4

Reference
1.57 (0.50, 4.92)
1.99 (0.62, 6.38)
1.94 (0.59, 6.35)

p for trend: 0.64
Total count (<39 M)

Q1
Q2
Q3
Q4

Reference
1.66 (0.51, 5.46)
3.33 (1.01, 10.99)
2.00 (0.58, 6.85)

p for trend: 0.24

Progressive motility (<32%)
Q1
Q2
Q3
Q4

Reference
4.36 (0.83, 22.81)
6.35 (1.21, 33.19)
2.40 (0.39, 14.49)

p for trend: 0.09

Viability (<58%)
Q1
Q2
Q3
Q4

Reference
8.00 (1.59, 40.30)
12.00 (2.34, 61.52)
10.15 (1.95, 52.92)

p for trend: 0.006

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

WHO morphology (<30%)

Q1
Q2
Q3
Q4

Reference
3.83 (0.924, 15.90)
6.57 (1.57, 27.43)
2.02 (0.426, 9.55)

p for trend: 0.06

Shi eta!. (2021)

Hong Kong

November 2015-
November 2016

Cross-sectional

n: 288

Male subjects who
underwent SA as part of
the fertility assessment at
the andrologyjaboratory
of Prince of Wales
Hospital. Participants were
excluded with medical
conditions azoospermia;
andrological conditions
(which are known to affect
semen parameters
including genetic
conditions); history of
mumps orchitis, severe
varicocele, undescended
testis; history of testicular
torsion or scrotal injury,
congenital bilateral
absence of vas deferent,
and urogenital infections;
taking medication known
to affect semen
parameters, including
steroid, finasteride,
calcium channel blockers;
history of malignant
disease; known mental
disorders; drug abuse;
failure to complete the

Blood

Blood was measured by
ICP-MS

Age at Measurement
Mean age: 37.9 yr

Geometric meanb:
3.175 [jg/dL
Medianb: 2.719 pg/dL
75thb: 3.437 [jg/dL

Quartilesb (pg/dL)

Q1
Q2
Q3
Q4

<2.159

>2.159-2.719
>2.719-3.437
>3.437

Male reproductive effects:
Seminal parameters

Semen volume was
measured by a wide-bore
graduated pipette_with the
graduation of 0.1-
ml. Sperm

concentration and motility
were examined under
a phase contrast
microscope with the
magnification of x 200 or
400. Diff-Quik staining kit
(Dade Behring AG,
DOdingen, Switzerland)
and Tygerberg Strict
Criteria were used to
evaluate the sperm
morphology. Sperm DNA
fragmentation was
measured by sperm
chromatin structure assay.

Age at outcome:

Mean age: 37.9 yr

Multivariate linear
regression adjusted for
(1) male age and daily
coffee intake for semen
volume models; (2)
abstinence time,
average sleep duration
for sperm concentration
models; (3) male age,
abstinence time, and
daily coffee intake for
total sperm count
models; (4) male age
and daily juice intake
for the sperm motility
models; (5) male age
and abstinence time for
total motility count
models; (6) no
adjustment for normal
morphology or sperm
vitality models; (7) male
age, abstinence time,
and irregular sleeping
habit for DNA
fragmentation index
models; (8) daily juice
intake for percentage of
acrosome reacted
sperm models.

B (95% CI)

Semen volume

Q1
Q2
Q3
Q4

Reference
-0.05 (-0.70, 0.37)
0.04 (-0.39, 0.65)
0.08 (-0.32, 0.83)

p for trend: 0.48

Sperm concentration

Q1
Q2
Q3
Q4

Reference
0.02 (-0.45, 0.58)
-0.02 (-0.57, 0.43)
-0.10 (-0.85, 0.26)

p for trend: 0.34

Total sperm count
Q1
Q2
Q3
Q4

Reference
-0.01 (-0.60, 0.53)
-0.03 (-0.66, 0.43)
-0.05 (-0.76, 0.44)

p for trend: 0.55

Sperm motility
Q1: Reference

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

lifestyle questionnaire; and
refusal to donate blood or
semen samples.

Q2
Q3
Q4

-0.09 (-10.21, 3.02)
-0.15 (-12.47, 0.49)
-0.08 (-10.26, 4.02)

p for trend: 0.77

Total motility count

Q1
Q2
Q3
Q4

Reference
-0.07 (-0.97, 0.38)
-0.08 (-0.97, 0.36)
-0.12 (-1.20, 0.25)

p for trend: 0.16

Normal morphology

Q1
Q2
Q3
Q4

Reference
-0.13 (-1.16, 0.13)
-0.20 (-1.43, -0.16)
-0.20 (-1.52, -0.10)

p for trend: 0.20

Sperm vitality
Q1
Q2
Q3
Q4

Reference
0.12 (-0.04, 0.17)
0.01 (-0.10, 0.12)
-0.13 (-0.19, 0.04)

p for trend: 0.13

Percentage of acrosome
reacted sperm
Q1
Q2
Q3
Q4

Reference
-0.22 (-18.60, 0.97)
-0.05 (-11.60, 7.71)
-0.12 (-15.70, 5.79)

p for trend: 0.75

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Pant eta!. (2014)

New Delhi
India

Cross-sectional

n: 60

Male partners of couples
age 21-40 yr old attending
the Andrology Laboratory
of the Reproductive
Biology Department, All
India Institute of Medical
Sciences, New Delhi,

India for semen analysis
to assess their inability to
achieve a pregnancy were
selected.

Other: Semen

Semen measured by ICP-
AES

Age at Measurement:
mean age: 31.81 (SD:
5.27)

Mean (SD): 6.18
(2.16) pg/dL

Male reproductive effects:
Semen quality

Semen of volunteers was
collected and analyzed the
protocols of the WHO.
Sperm morphology was
determined according to
Kruger's strict criteria.
Comet assay: prepared
sperm samples were
observed under a
fluorescence microscope
with a total of 100 cells
were scored. The
percentage of tail DNA, tail
length, and tail moment
was evaluated by the
CometScore software
image analysis system.

Age at outcome:
mean age: 31.81 (SD:
5.27)

Multiple regressions,
adjusted for toxicants
(Cd, diethyl phthalate,
dibutyl phthalate, di[2-
ethylhexyl] phthalate),
age, BMI, tobacco,
smoking, alcohol, and
diet

(3 (95% Cl)c

Sperm motility (%): 2.43
(-4.87, -0.001)

Sperm concentration
(106/ml): -1.97 (-3.16,
-0.33)

Tail length: 3.79 (0.56, 7.02)

Percent DNA in tail: 1.31
(0.172, 3.74)

Tail moment: 1.20 (0.23,
2.16)

Jia et al. (2022)

Henan Province
China

December 2017 to
August 2018

Cross-sectional

n: 841

Males ranging from 18 to
50 yr of age with no
history of testicular injury,
urologist diagnosed
inflammation of the
urogenital system; history
of epididymitis; treatment
history of varicocele;
history of incomplete
orchiocatabasis or any of
the following that was
detected by an urologist at
physical examination:

Other: Semen

Seminal plasma was
measured by analyzed
using the kinetic energy

discrimination-based
Thermo iCAP Q ICP-MS

Age at Measurement

Mean ± SD:

29.55 ±5.45 yr

Median: 1.70 ppb

Male reproductive effects:
Seminal parameters

Semen of volunteers was
collected and analyzed the
protocols of the WHO.
Computer-assisted sperm
analysis technology was
used to analyze the
collected semen samples.
The quality indicators were
complete liquefaction,
semen volume, sperm
concentration, total sperm
count, progressive motility,

Multilinear regression
models were adjusted
for age, BMI, smoking,
and alcohol
consumption

(3 (95% Cl)c, per increase in
In-Pb seminal plasma
Semen volume: -0.10
(-0.27, 0.07)

Sperm concentration: 1.83
(-4.45, 8.12)

Total sperm number: 0.80
(-17.61, 19.21)

Progressive motility: 0.06
(-2.09, 2.21)

Normal morphological rate:
-0.04 (-0.41, 0.34)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

absence of prominentia
laryngea, absence of
pubes, abnormal breast,
absence of testis,
abnormal penis,
epididymal knob, or
varicocele.

75th: 2.36 ppb

non-progressive motility,
sperm motility, and sperm
motility parameters, such
as curve line velocity
(|jm/s), straight line
velocity (|-im/s), velocity of
average path (|-im/s),
lateral head movement
(amplitude of lateral head
displacement, |jm),
average motion degree (°),
linearity (%), straightness
(%), wobble, and beat
cross frequency (beat
cross frequency, Hz).

Curve line velocity: 0.35
(-1.17, 1.88)

Straight line velocity: 0.54
(-0.50, 1.58)

Velocity of average path:
0.37 (-0.96, 1.70)

Linearity: 0.49 (-0.88, 1.86)

Straightness: 0.46 (-1.07,
1.99)

Wobble: 0.22 (-1.33, 1.77)

Average motion degree:
-0.33 (-1.22, 0.56)

Beat cross frequency: 0.01 (-
0.14, 0.15)

Lateral head movement:
-0.04 (-0.11, 0.03)

Williams et al. (2022) Russian Children's Study Blood
n: 223

Russia

Boys enrolled at age 8-
2003-2005 (follow- 9 yr in 2003-2005 and
up annually for	followed them annually for GFAAS

10 yr)	10 yr.

Blood was measured
Zeeman background
corrected flameless

by

Cohort

Age at measurement:
9 yr

Median: 3 [jg/dL
75th: 5 [jg/dL

Categories
Lower: <5 [jg/dL
Higher: >5 [jg/dL

Tertiles

Male reproductive effects:
Semen parameters

All semen samples were
assessed by a single
andrology technician and
analyzed according to
criteria of the Nordic
Association for

Andrology and European
Society of Human
Reproduction and
Embryology-Special
Interest Group in
Andrology and serum
hormonal levels were
analyzed using the
Architect i1000SR and
chemiluminescent

Mixed effect linear
regression models
adjusted for boys' BW,
total caloric intake, HTZ
at entry, breastfeeding
duration, monthly
household income, and
abstinence time

(3 (95% Cl)b, as adjusted
mean

Semen volume (mL)

Continuous, per log-blood
Pb: -0.40 (-0.82, 0.03)

Categories

Lower: 2.83 (2.61, 3.06)
Higher: 2.60 (2.27, 2.93)
Tertiles

Low: 2.92 (2.50, 3.34)
Medium: 2.79 (2.52, 3.06)
High: 2.60 (2.27, 2.93)
p for trend: 0.24

Sperm concentration
(mill/mL)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Low: <2 [jg/dL
Medium: 3-4 [jg/dL
High: >5 [jg/dL

microparticle
immunoassay.

Age at outcome: 18 yr or
older

Continuous, per log-blood
Pb: 0.09 (-0.13, 0.31)

Categories

Lower: 47.0 (41.3, 53.4)
Higher: 49.0 (37.8, 63.4)
Tertiles

Low: 41.3 (33.2, 51.3)
Medium:50.3 (42.8, 59.0)
High: 49.1 (38.0, 63.6)
p for trend: 0.33

Total sperm count (mill)

Continuous, per log-blood
Pb: -0.02 (-0.27, 0.23)

Categories

Lower: 111 (95.6, 129)
Higher: 107 (80.0, 143)
Tertiles

Low: 99 (76.4, 128)
Medium: 118 (95.5, 141)
High: 107 (80.3, 143)
p for trend: 0.68

Progressive sperm motility

(%)

Continuous, per log-blood
Pb: 1.77 (-0.55, 4.08)

Categories

Lower: 53.2 (51.7, 54.7)
Higher: 53.1 (50.9, 55.2)
Tertiles

Low: 51.2 (48.6, 53.9)
Medium: 54.3 (52.5, 56.1)

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

High: 53.2 (50.9, 55.3)
p for trend: 0.29

Effects of Hormone Levels

Total progressive motile
sperm count (mill)

Continuous, per log-blood
Pb: 0.01 (-0.27, 0.29)

Categories

Lower: 57.7 (48.9, 68.1)
Higher: 55.7 (40.6, 76.4)
Tertiles

Low: 49.4 (36.8, 66.2)
Medium: 62.6 (51.3, 76.5)
High: 56.0 (40.8, 76.8)
p for trend: 0.57

Low semen quality
(probability)

Continuous, per log-blood
Pb: 0.20 (-0.22, 0.65)
Categories

Lower: 0.51 (0.44, 0.58)
Higher: 0.49 (0.39, 0.59)
Tertiles

Low: 0.43 (0.31, 0.55)
Medium: 0.55 (0.46, 0.63)
High: 0.49 (0.39, 0.59)
p for trend: 0.43

Kresovich et al. NHANES	Blood	Male reproductive effects: Linear regression	(3 (SE)d

n: 869	Hormones	models were adjusted	Testosterone (ng/mL)

for age, BMI, race,	„ *

a	Q1: Reference

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

United States

1999-2004

Cross-sectional

Males who were aged
>20 yr, no reported steroid
or thyroid mediation use,
and no reported thyroid
disease.

Blood was measured by
AAS (1999-2002) or ICP-
MS (2003-2004).

Age at measurement:
>20 yr

Median (weighted):
2.0 [jg/dL
75th: 2.8 pg/dL

Quartiles (pg/dL)

Testosterone,
androstanedione
glucuronide, and SHBG
were measured in blood
serum, and E2 in plasma.
All sex hormones were
detected by immunoassay.

Age at outcome: >20 yr

diabetes status
(including prediabetes),
smoking status, and
alcohol intake; and Cd

Q1
Q2
Q3
Q4

<1.40
1.40-2.10
2.10-3.20
>3.20

Q2
Q3
Q4

0.39 (0.21)
0.56 (0.22)
0.81 (0.20)

p for trend: 0.0008

E2 (pg/mL)

Q1
Q2
Q3
Q4

Reference
-0.01 (0.03)
-0.01 (0.04)
-0.01 (0.04)

p for trend: 0.7849

IT (ng/dL)

Q1
Q2
Q3
Q4

Reference
0.83 (0.47)
0.55 (0.48)
0.81 (0.48)

p for trend: 0.2374

fE2 (pg/ml)

Q1
Q2
Q3
Q4

Reference
-0.01 (0.03)
-0.02 (0.04)
-0.03 (0.04)

p for trend: 0.4428

Androstanedione
glucuronide (ng/mL)
Q1: Reference
Q2: 0.03 (0.03)
Q3: -0.01 (0.03)
Q4: 0.02 (0.04)
p for trend: 0.8917

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Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

SHBG (nmol/L)

Q1
Q2
Q3
Q4

Reference
0.01 (0.02)
0.05 (0.02)
0.05 (0.02)

p for trend: 0.0187

Adjusted for Cd
Testosterone (ng/mL)
Q1
Q2
Q3
Q4

Reference
0.38 (0.23)
0.54 (0.21)
0.79 (0.22)

p for trend: 0.0026

E2 (pg/mL)

Q1
Q2
Q3
Q4

Reference
0.00 (0.03)
0.01 (0.04)
0.02 (0.04)

p for trend: 0.6600

IT (ng/dL)

Q1
Q2
Q3
Q4

Reference
0.95 (0.50)
0.70 (0.51)
1.06 (0.51)

p for trend: 0.1388

fE2 (pg/ml)
Q1: Reference
Q2: 0.01 (0.03)

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Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Q3: 0.00 (0.04)
Q4: 0.01 (0.04)
p for trend: 0.9456

Androstanedione
glucuronide (ng/mL)
Q1: Reference
Q2: 0.03 (0.03)
Q3: -0.02 (0.03)
Q4: 0.01 (0.04)
p for trend: 0.7620

SHBG (nmol/L)
Q1: Reference
Q2: -0.01 (0.02)
Q3: 0.03 (0.02)
Q4: 0.03 (0.03)
p for trend: 0.1333

Lewis and

United States

2011-2012

Cross-sectional

NHANES
n: 484

Men that were 18-55 yr
old, that had complete
data on the metals of
interest, serum
testosterone, BMI, PIR,
race, serum cotinine, or
urinary creatinine

Blood

Blood was measured by
inductively coupled
dynamic reaction-plasma
mass spectrometry

Age at Measurement:
18-55yr

Geometric mean:
1.06 [jg/dL
75th: 1.59 pg/dL

Male reproductive effects:
Testosterone

Serum testosterone (total)
were measured by isotope
dilution-high performance
liquid chromatography-
tandem mass
spectrometry

Age at outcome:

18-55 yr

Multiple linear
regression, adjusted for
age, BMI, PIR, race,
and serum cotinine

(3 (95% CI)c, as percent
change in serum
testosterone associated with
a doubling (100% increase)
in blood Pb concentration:
6.65 (2.09, 11.41)

Chen et al. (2016) SPECT-China	Blood	Male reproductive effects: Linear regression	(3 (SE)d

n: 2286 men	Reproductive hormone models were adjusted SHBG

Shanghai, Jiangxi	levels	for age and current

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Province and
Zhejiang province
China

2014

Cross-sectional

SPECT-China is a
population-based cross-
sectional survey on the
prevalence of metabolic
diseases and risk factors
in East China. Men and
postmenopausal women
(age >55 yr) who were not
taking hormone
replacement therapy,
without a history of
hysterectomy and
oophorectomy were
recruited.

Blood was measured by
AAS

Age at Measurement:
Median (IQR) age: 54
(44-63)

Medianb: 4.400 |jg/dL
75thb: 6.230 pg/dL

Quartilesb (pg/dL)

Q1
Q2
Q3

<2.900

2.900-4.399

4.400-6.229

Q4: >6.229

Venous blood samples
were drawn from all
subjects after an overnight
fast of at least 8 hr. HbA1c
was assessed via high-
performance liquid
chromatography. tT, E2,
LH and FSH levels were
measured using
chemiluminescence
assays. SHBG levels were
detected using
electrochemiluminescence
immunoassays.

Age at outcome:

Median (IQR) age: 54 (44-

63)

smoking status, BMI,
SBP, diabetes and,
blood Cd level

Q1
Q2
Q3
Q4

Reference
<0.001 (0.011)
0.021 (0.011)
0.038 (0.012)

p for trend: <0.001

tT

Q1

Q2

Q3

Q4

Reference
0.001 (0.010)
0.010 (0.010)
0.033 (0.010)

p for trend: 0.001

E2
Q1
Q2
Q3
Q4

Reference
-0.008 (0.016)
0.014 (0.017)
-0.003 (0.017)
p for trend: 0.794

FSH

Q1: Reference
Q2: 0.010 (0.014)
Q3: 0.004 (0.014)
Q4: 0.030 (0.015)
p for trend: 0.067

LH

Q1: Reference
Q2: 0.018 (0.013)
Q3: 0.015 (0.013)
Q4: 0.028 (0.013)
p for trend: 0.065

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

Effects on Fertility

Louis et al. (2012)

Michigan (4
counties) and Texas
(12 counties)

United States

2005-2009

Cohort

LIFE Study
n: 501

Female ages 18-44 yr
and male ages >18 yr; in a
committed relationship;
ability to communicate in
English or Spanish;
menstrual cycles between
21 and 42 d; no hormonal
contraception injections
during past year; and no
sterilization procedures or
physician diagnosed
infertility

Blood

Blood was measured by
ICP-MS

Age at Measurement:
Average age for male
partner with pregnancy:
31.6 yr

Average age for male
partner without
pregnancy: 32.4 yr

Geometric mean

Male partner with
pregnancy result:
1.03 [jg/dL

Male partner without
pregnant result:
1.18 pg/dL

Male reproductive effects:
Fecundity

Women were instructed in
the use of the Clearblue
Easy fertility monitors
consistent with the
manufacturer's guidance
commencing on day six for
tracking daily levels of
E3G and LH. Women also
used the digital Clearblue
Easy home pregnancy test
upon enrollment to ensure
the absence of pregnancy
at study start and on the
day menses was expected
for each cycle under
observation in the study.

Age at outcome:

Average age for males
with pregnancy: 31.6 yr
Average age for males
without pregnancy: 32.4 yr

Cox models for discrete
survival time, which is a
proportional odds
model, adjusted for
age, BMI, cotinine,
parity, serum lipids, and
site (Texas/Michigan)

OR (95% CI), as
fecundability OR

Male only exposure: 0.85
(0.73, 0.99)

Couple exposure:

Female exposure: 1.06
(0.91, 1.24)

Male exposure: 0.82 (0.68,
0.97)

Zhou et al.

China

2018-2019

Cohort

n: 195

Couples undergoing IVF.
Women with
endometriosis,
hydrosalpinx, abnormal
uterine cavity and men
with azoospermia, severe
oligozoospermia,
asthenospermia and

Blood, other: follicular
fluid, and other: semen

Maternal blood (serum),
follicular fluid, and
seminal plasma from
male partner

Age at Measurement:

Male reproductive effects:
IVF outcome

The IVF outcomes
included were normal
fertilization, good embryo,
blastocyst formation, high-
quality blastocyst,
pregnancy, and live birth

Age at outcome:

Poisson regression
models were adjusted
for age and BMI

RR (95% Cl)c
Normal fertilization
Maternal serum: 0.94 (0.42,
1.93)

Follicular fluid: 0.82 (0.18,
2.39)

Seminal plasma: 1.55 (0.64,
3.3)

Good embryo

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Reference and
Study Design

Study Population

Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

dysspermia were excluded Female partner mean
from the study.	age: 30.27 yr

Male partner mean age:
31.57 yr

Mean0

Maternal serum:
0.301 [jg/dL
Follicular fluid:
0.742 [jg/dL
Seminal plasma:
0.882 [jg/dL

Median0

Maternal serum:
0.245 [jg/dL
Follicular fluid:
0.178 [jg/dL
Seminal plasma:
0.486 [jg/dL

75th°

Maternal serum:
0.317 [jg/dL
Follicular fluid:
0.326 [jg/dL
Seminal plasma:
1.245 [jg/dL

Female partner mean age:
30.27 yr

Male partner mean age:
31.57 yr

Maternal serum: 1.00 (0.36,
2.38)

Follicular fluid: 0.78 (0.09,
3.03)

Seminal plasma: 1.86 (1.05,
3.11)

Blastocyst formation

Maternal serum: 1.06 (0.2,
3.91)

Follicular fluid: 0.41 (0, 3.63)
Seminal plasma: 1.77 (0.78,
3.58)

High-quality blastocyst

Maternal serum: 1.68 (0.15,
9.43)

Follicular fluid: 0.35 (0, 7.11)
Seminal plasma: 2.66 (0.67,
8)

Pregnancy

Maternal serum: 0.18 (0.01,
1.91)

Follicular fluid: 0.01 (0, 0.03)

Seminal plasma: 0.04 (0,
1.45)

Live birth

Maternal serum: 0.25 (0.01,
2.8)

Follicular fluid: 0 (0, 0.09)

Seminal plasma: 0.01 (0,
1.08)

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study Desfgn	Study P°Pulation ExP°sure Assessment	Outcome	Confounders Effect Esti™£s and 95%

Effects on Morphology or Histology of Male Sex Organs

Huang et al.

Guangxi
China

July 2015 to
September 2018

Cohort

Guangxi Birth Cohort
Study

n: 564 mother-child pairs

Women with singleton
pregnancies that were
included from 8 Maternity
and Child Healthcare
Hospitals in 6 cities of
Guangxi, China

Blood

Maternal blood (serum)
was measured by ICP-
MS

Age at Measurement:
Maternal age at time of
measurement (mean age:
28.76 (SD: 4.66) yr)

Medianb: 0.077 [jg/dL
75thb: 0.123 pg/dL

Quartilesb (pg/dL)

Q1
Q2
Q3
Q4

<0.054
0.055-0.077
0.078-0.123
>0.123

Male reproductive effects:
TV and AGD in infant boys

TV, and AGD-TV
measurements were
undertaken by trained
sonographers using
ultrasonography.
Transverse and
longitudinal grey-scale
images were used to
calculate TV as
tt/6 x length * width * heig
ht. The volumes of both
testes were measured and
an average taken. Two
different measurements of
AGD were obtained using
vernier calipers: the longer
AGD was measured from
the center of the anus to
the cephalad insertion of
the penis (AGDap), and
the shorter AGD was
measured from the center
of the anus to the posterior
base of the scrotum
(AGDas).

Age at outcome:
birth

Multiple linear

(3 (95% Cl)c

regression models were yy
adjusted for BW, GA,
blood sampling time
(mother), alcohol use
pre-pregnancy, BMI,
and age at examination

Q1
Q2
Q3
Q4

Reference

-0.017 (-0.077, 0.043)
-0.024 (-0.085, 0.036)
-0.064 (-0.124, -0.004)

AGDap
Q1
Q2
Q3
Q4

Reference

-0.039 (-0.085, 0.008)
-0.037 (-0.085, 0.010)
-0.060 (-0.110, -0.011)

AGDas

Q1
Q2
Q3
Q4

Reference

-0.020 (-0.091, 0.052)
-0.033 (-0.105, 0.039)
-0.115 (-0.190, -0.039)

AAS = atomic absorption spectrometry; AGD = anogenital distance; AGDap = anopenile distance; AGDas = anoscrotal distance; BMI = body mass index; BW = birth weight;
CI = confidence interval; d = day(s); E2 = estradiol; E3G = estrone-3-glucuronide; fE2 = free estradiol; FSH = follicle stimulating hormone; fT = free testosterone; FSH = follicle
stimulating hormone; GA = gestational age; GFAAS = graphite furnace atomic absorption spectrometry; HTZ = height Z-score; ICP-AES = inductively coupled plasma atomic
emission spectrometry; ICP-MS = inductively coupled plasma mass spectrometry; IVF = in vitro fertilization; LH = luteinizing hormone; LIFE = Longitudinal Investigation of Fertility
and the Environment; LOD = limit of detection; mo = month(s); NHANES = National Health and Nutrition Examination Survey; OR = odds ratio; PIR = poverty-income ratio;
Q = quartile; SA = semen analysis; SBP = systolic blood pressure; SD = standard deviation; SE = standard error; SHBG = sex hormone binding globulin; SPECT = Survey on the
Prevalence in East China for Metabolic Diseases and Risk Factors; T = testosterone; tT = total testosterone; TV = testicular volume; WHO = World Health Organization; yr = year(s).

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Reference and
Study Design

Study Population Exposure Assessment

Outcome

Confounders

Effect Estimates and 95%
Clsa

aEffect estimates are standardized to a 1 |jg/dL increase in blood Pb or a 10 |jg/g increase in bone Pb, unless otherwise noted. If the Pb biomarker is log-transformed, effect

estimates are standardized to the specified unit increase for the 10th—90th percentile interval of the biomarker level. Effect estimates are assumed to be linear within the evaluated

interval. Categorical effect estimates are not standardized.

bPb measurements were converted from |jg/L to |jg/dL.

°Effects estimates unable to be standardized.

dNo CIs provided.

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Table 8-17

Animal toxicological studies of exposure to Pb and male reproductive effects

Study

Species (Stock/Strain), n, Sex

Timing of
Exposure

Exposure Details
(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

El Shafai et al. (2011) Rat (Wistar)

Control (untreated), M, n = 8

Control (vehicle), M, n = 8

25 mg/kg Pb, M, n = 8

Adulthood (specific
PND NR)

Adult male rats were
dosed via oral gavage for
3 mo. One control group
was not gavaged
(untreated control) and
another control group
was gavaged with vehicle
(vehicle control).

4.26	[jg/dL for
control (untreated)

4.27	[jg/dL for
control (vehicle)

5.27 [jg/dL for
25 mg/kg Pb

Sex Organ
Histopathology

Wang et al. (2013b)

Rat (Sprague-Dawley)

Control (untreated), M, n = 15

0.8/0.3 g/L Pb, M, n = 15
1.5/0.9 g/L Pb, M, n = 15

GD -10 to	Dams were dosed via

PND 183	drinking water (0, 0.8, or

1.5 g/L Pb) starting 10 d
prior to mating through
weaning. At weaning 15
males from each group
were dosed via drinking
water to lower levels of
Pb than their dams (0,
0.3, or 0.9 g/L) until 6 mo
of age (approx.
PND 183).

2.65 [jg/dL for
control

18.6 [jg/dL for
0.8/0.3 g/L Pb

55.0 [jg/dL for
1.5/0.9 g/L Pb

Testicular
Weight

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Study

Species (Stock/Strain), n, Sex

Timing of
Exposure

Exposure Details
(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Wang et al. (2013a)

Mouse (CD-1)

Control (untreated), M, n = 12
200 ppm Pb, M, n = 12
2000 ppm Pb, M, n = 12

PNDOto PND21

Dams were dosed via
drinking water from
PNDOto 21.

Pups:

PND 22

17.4 [jg/dL for
control

21.2 [jg/dL for
200 ppm Pb

19.1 [jg/L for
2000 ppm Pb

Testosterone
Levels, Sex
Organ

Histopathology,
Accessory Male
Reproductive
Organ Weight,
Testicular
Weight, Semen
Parameters

PND 70

4.40 [jg/dL for
control

3.24 [jg/dL for
200 ppm Pb

5.09 [jg/dL for
2000 ppm Pb

Godfnez-Solis et al.

Mouse (ICR-CD-1)

Control (untreated), M, n = 4

0.01% Pb, M, n = 6

PND 91 to 136

12 wk old mice were
acclimated for a week
before being dosed via
drinking water for 45 d.

BLL NR for controls

9.4 [jg/dL for
0.01% Pb

Semen

Parameters,

Sperm

Morphology, IVF

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Study

Species (Stock/Strain), n, Sex

Timing of
Exposure

Exposure Details
(Concentration,
Duration)

BLL as Reported
(Hg/dL)

Endpoints
Examined

Xie et al. (2020)

Mouse (SPF ICR)

Control (untreated), M, n = 15

50 mg/L Pb, M, n = 15
200 mg/L Pb, M, n = 15

PND 28 to
PND 118

21 d old mice were
acclimated for a week
before being dosed for
90 d via drinking water.

0.602 |jg/dL for
control

6.02 |jg/dL for
50 mg/L Pb

11.8 |jg/dL for
200 mg/L Pb

Semen

Parameters,

Sperm

Morphology,

Sex Organ

Histopathology,

Testicular

Weight,

Accessory Male
Reproductive
Organ Weight

Pavlova et al, (2021)

Mouse (ICR)

Control (vehicle), M, n = 10
80 mg/kg Pb, M, n = 10

PND 60 to 74

60 d old mice were
dosed via oral gavage for
2 wk. Two weeks
following cessation of
exposure, animals were
sacrificed.

1.45 |jg/dL for
control

21.66 |jg/dL for
80 mg/kg Pb

Testicular
Weight, Semen
Parameters, Sex
Organ

Histopathology

BLL = blood lead level; d = day(s); F = female; GD = gestational day; IVF = in vitro fertilization; M = male; mo = month(s); NR = not reported; Pb = lead; PND = postnatal day;
T = testosterone; wk = week(s).

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8.10 References

Abdelhamid. FM; Mahgoub. HA; Ateva. AI. (2020). Ameliorative effect of curcumin against lead acetate-induced
hemato-biochemical alterations, hepatotoxicity, and testicular oxidative damage in rats. Environ Sci Pollut
Res Int 27: 10950-10965. http://dx.doi.org/10.1007/sll356-020-Q7718-3.

Abdrabou. MI: EHeithv. EMM: Yasin. NAE: Shaheen. YM: Gala I M. (2019). Ameliorative effects of Spirulina
maxima and Allium sativum on lead acetate-induced testicular injury in male albino rats with respect to
caspase-3 gene expression. Acta Histochem 121: 198-206. http://dx.doi.Org/10.1016/i.acthis.2018.12.006.

Afeiche. M; Peterson. KE; Sanchez. BN; Schnaas. L; Cantonwine. D; Ettinger. AS; Sotano-Gonzalez. M;

Hernandez-Avila. M; Hu. H; Teltez-Rojo. MM. (2012). Windows of lead exposure sensitivity, attained
height, and body mass index at 48 months. J Pediatr 160: 1044-1049.

http://dx.doi.org/10. .1.0.1.6/i.jpeds.2011.1.2.022.

Al~Saleh. I; Shinwari. N; Mashhour. A; Rabah. A. (2014). Birth outcome measures and maternal exposure to heavy
metals (lead, cadmium and mercury) in Saudi Arabian population. Int I Hyg Environ Health 217: 205-218.

http://dx.doi.org/10. .1.01.6/i. iiheh.20.1.3.04.009.

Albores-Garcia. D; L McGlothan. J; Bursae. Z; Guilarte. TR. (2021). Chronic developmental lead exposure
increases (i-opiate receptor levels in the adolescent rat brain. Neurotoxicology 82: 119-129.

http://dx.doi.Org/.l.0.1016/i.nenro.2020..1.1.008.

Alegria-Torres. IA; Perez-Rodriguez. RY; Garcia-Torres. L; Costilla-Salazar. R; Rocha-Amador. D. (2020).

Exposure to arsenic and lead in children from Salamanca Mexico, effects on telomeric lengthening and
mitochondrial DNA. Environ Sci Pollut Res Int 27: 6420-6428. http://dx.doi.org/10.1007/sll356~019~
07.1.08-4.

Allonche. L; Hamadouche. M; Touabti. A. (2009). Chronic effects of low lead levels on sperm quality,
gonadotropins and testosterone in albino rats. Exp Toxicol Pathol 61: 503-510.

http://dx.doi.org/10. .1.0.1.6/i.etp.2008.1.2.003.

Alotalbi. MF; Al-Jonfi. F; Abou Seif. HS; Alzoghaibi. MA; Djonhri. L; Ahmeda. AF; Mahmond. AM. (2020).

Umbelliferone inhibits spermatogenic defects and testicular injury in lead-intoxicated rats by suppressing
oxidative stress and inflammation, and improving Nrf2/HO-l signaling. Drug Design, Development and
Therapy 14: 4003-4019. http://dx.doi.org/10.2147/DDDT.S265636.

Aniiim. KM; Sainath. SB; Suneetha. Y; Reddv. SP. (2011). Lead acetate induced reproductive and paternal mediated
developmental toxicity in rats. Ecotoxicol Environ Saf 74: 793-799.

http://dx.doi.org/.1.0..1.0.1.6/i.ecoenv.201.0..1.0.044.

Ashley-Mart in. J; Dodds. L; Arbtickle. TE; Ettinger. AS; Shapiro. GD; Fisher. M; Taback. S; Bouchard. MF;

Monnier. P; Dattalre. R; Fraser. WD. (2015a). Maternal blood metal levels and fetal markers of metabolic
function. Environ Res 136: 27-34. http://dx.doi.Org/10.1016/i.envres.2014..1.0.024.

Ashley-Mart in. J; Dodds. L; Arbtickle. TE; Lanphear. B; Mnekle. G; Bouchard. MF; Fisher. M; Asztalos. E; Foster.
W; Knlile. S. (2019). Blood metal levels and early childhood anthropometric measures in a cohort of
Canadian children. Environ Res 179: 108736. http://dx.doi.org/10. .1.01.6/i.envres.201.9. .1.08736.

Ashley-Mart in. J; Levy. AR; Arbuckle. TE; Piatt. RW; Marshall. IS; Dodds. L. (2015b). Maternal exposure to
metals and persistent pollutants and cord blood immune system biomarkers. Environ Health 14: 52.

http://dx.doi.org/10. .1.186/sl.2940~0.1.5-0046-3.

8-243


-------
Ashrap. P; Aker. A; Watkins. DJ; Miikheriee. B; Rosario-Pabon. Z; Velez-Vega. CM; Alshawabkeh. A; Cordero.
JF; Meeke . (2021). Psychosocial status modifies the effect of maternal blood metal and metalloid
concentrations on birth outcomes. Environ Int 149: 106418.

http://dx.doi.org/10. .1.016/i .envint.202.1.. .1.064.1.8.

Ashrap. P; Watkins. DJ; Mukherjee. B; Boss. J; Richards. MI; Rosario. Z; Velez-Vega. CM; Alshawabkeh. A;

Cordero. JF; Meek . (2020). Maternal blood metal and metalloid concentrations in association with
birth outcomes in Northern Puerto Rico. Environ Int 138: 105606.

http://dx.doi.Org/10.1016/i.envint.2020..1.05606.

Baranowska-Bosiacka. I; Struzvnska. L; Gutowska. I; Maehatinska. A; Kolasa. A; Klos. P; Czapski. GA;

Kurzawski. M; Prokopowicz. A; Marchlewicz. M; Safranow. K; Maehatinski. B; Wiszniewska. B;

Chlubek. D. (2013). Perinatal exposure to lead induces morphological, ultrastructural and molecular
alterations in the hippocampus. Toxicology 303: 187-200. http://dx.doi.Org/10.1016/i.tox.2012.10.027.

Barkur. RR; Bairv. LK. (20.1.5). Evaluation of passive avoidance learning and spatial memory in rats exposed to low
levels of lead during specific periods of early brain development. Int J Occup Med Environ Health 28: 533-
544. http://dx.doi.Org/.l.0.13075/iiomeh.1896.00283.

Barkur. RR; Rao. MS; Bairv. LK. (20.1.1). Low lead exposure during foetal and early postnatal life impairs passive
avoidance learning in adulthood in rats. Arh Hig Rada Toksikol 62: 147-153.

http://dx.doi.org/10.2478/10004-1254-62-2011-2070.

BaSalamah. MA; Abdelghany. AH; El-Boshy. M; Ahmad. J; Idris. S; Refaat. B. (2018). Vitamin D alleviates lead
induced renal and testicular injuries by immunomodulatory and antioxidant mechanisms in rats. Sci Rep 8:
4853. http://dx.doi.org/10.1038/s41598-Q18-23258-w.

Basgen. JM; Sob in. C. (20.1.4). Early chronic low-level lead exposure produces glomerular hypertrophy in young
C57BL/6J mice. Toxicol Lett 225: 48-56. http://dx.doi.org/10. .1.016/i.toxlet.20.1.3. .1. .1..03.1..

Basha. CD; Reddv. RG. (20.1.5). Long-term changes in brain cholinergic system and behavior in rats following
gestational exposure to lead: Protective effect of calcium supplement. Interdiscip Toxicol 8: 159-168.
htt p ://dx. do i.org/ .1.0. .1.5.1.5/i ntox-20.1.5-0025.

Betharia. S; Maher. TJ. (20.1.2). Neurobehavioral effects of lead and manganese individually and in combination in
developmentally exposed rats. Neurotoxicology 33: 1117-1127.

htt p ://dx. do i .o rg/ .1.0. .1.0.1.6/i. neuro.20.1.2.06.002.

Biswas. NM; Ghosh. PK. (2006). Protection of adrenal and male gonadal functions by androgen in lead-treated rats.
Kathmandu Univ Med J 4: 218-221.

Bloom. MS; Buck Louis. GM; Sundarani. R; Maisog. JM; Stenerwald. AJ; Parsons. PJ. (2015). Birth outcomes and
background exposures to select elements, the Longitudinal Investigation of Fertility and the Environment
(LIFE). Environ Res 138: 118-129. http://dx.doi.Org/.l.0.10.l.6/i.envres.2015.0L008.

Brann. JM; Wright. RJ; Just. AC; Power. MC; Ortiz. MTY; Sehnaas. L; Hn. H; Wright. RO; Teltez-Rojo. MM.

(2014). Relationships between lead biomarkers and diurnal salivary Cortisol indices in pregnant women
from Mexico City: Across-sectional study. Environ Health 13: 50. http://dx.doi.org/10. .1.186/1476-069X-

.1.3-50.

Bui. LTM; Shadbegian. R; Marquez. A; Klemick. H; Guignet. D. (2022). Does short-term, airborne lead exposure
during pregnancy affect birth outcomes? Quasi-experimental evidence from NASCAR's deleading policy.
Environ Int 166: 107354. http://dx.doi.Org/10.1016/i.envint.2022..1.07354.

Bums. JS; Williams. PL; Lee. MM; Revich. B; Sergeyev. O; Hauser. R; Korrick. SA. (2017). Peripubertal blood
lead levels and growth among Russian boys. Environ Int 106: 53-59.

http://dx.doi.Org/10.1016/i.envint.20.1.7.05.023.

Cass idy -Bushrow. AE; Wn. KHH; Sitarik. AR; Park. SK; Bielak. LF; Austin. C; Gennings. C; Curt in. P; Johnson.
CC; Arora. M. (2019). In utero metal exposures measured in deciduous teeth and birth outcomes in a
racially-diverse urban cohort. Environ Res 171: 444-451. http://dx.doi.Org/10.1016/i.envres.2019.01.054.

8-244


-------
Chen. C; Wang. NJ; Ziiai. HL; Nie. XM: Sun. HL; Han. 6; Li. O; Chen. Y; Cheng. J; Xia. FZ: Zhao. L; Zheng. YJ;
Shen. ZJ; Lu. YL. (2016). Associations of blood lead levels with reproductive hormone levels in men and
postmenopausal women: Results from the SPECT-China Study. Sci Rep 6: 37809.
http://dx.doi.org/10.1038/srep378Q9.

Choi. J: Chang	ng. J: Shin. S: Park. IS: Oh. S. (2017). Low-level toxic metal exposure in healthy weaning-

age infants: Association with growth, dietary intake, and iron deficiency. Int J Environ Res Public Health
14: 388. http://dx.doi.Org/.l.0.3390/iierph.l.4040388.

Christensen. PS: Bonde. .IP: Bungiim. L; Giwercman. A: Toft. G: Jonsson. BAG: Speeht. 10. (2016). Environmental
cadmium and lead exposure and anti-Mullerian hormone in pregnant women. Reprod Toxicol 61: 114-119.

http://dx.doi.Org/.l.0.1016/i.reprotox.20.l.6.03.047.

Corrales Varga. \ l Baloza Castaneda. J: Rietz Liljedahl. E: Mora. AM: Menezes-Fillw i \ "'mith. PR: Mergler.
D: Reich. B: Giffin. A: Hoppin.	Ah. CH: van Wendel de Joode. B. (2022). Exposure to common-use

pesticides, manganese, lead, and thyroid function among pregnant women from the Infants' Environmental
Health (ISA) study, Costa Rica. Sci Total Environ 810: 151288.

htt p ://dx. do i. o rg/ .1.0. .1.0.1.6/i. sc i to t e nv.202.1. .151288.

Corv-Slechta. DA: Merchant-Borna. K: Allen. JL: Lin. S: Weston. D: Conrad. K. (2013). Variations in the nature of
behavioral experience can differentially alter the consequences of developmental exposures to lead,
prenatal stress, and the combination. Toxicol Sci 131: 194-205. http://dx.doi.Org/.l.0.1093/toxsci/kfs260.

Ciillen. C: Singh. A: Dykeman. A: Rice. D: Foster. W. (1993). Chronic lead exposure induces ultrastructural
alterations in the monkey seminal vesicle. J Submicrosc Cytol Pathol 25: 127-135.

Daflarv. SS: Gore. AC. (2005). IGF-1 in the brain as a regulator of reproductive neuroendocrine function [Review].
Exp Biol Med 230: 292-306. http://dx.doi.or;	3537020523000503.

Daniali. SS: Yazdi. M: Heidari-Beni. M: Taheri. E: Zarean. E: Goti. P: Kelishadi. R. (2023). Birth size outcomes in
relation to maternal blood levels of some essential and toxic elements. Biol Trace Elem Res 201: 4-13.

http://dx.doi.org/10.1007/sl201.1.-022-03.1.2.1.-w.

De Craetner. S: Croes. K: van Larebeke. N: De Henauw. S: Schoeters. G: Govarts. E: Loots. I: Nawrot. T: Nelen. V:
Den Hond. E: Bnickers. L: Gao. Y: Baevens. W. (2017). Metals, hormones and sexual maturation in
Flemish adolescents in three cross-sectional studies (2002-2015). Environ Int 102: 190-199.

http://dx.doi.Org/10.1016/i.envint.20.1.7.02.0.1.4.

de Figueiredo. FAT: Gerlach. RF: da Veiga. MAM. S: Nakadi. FY: Ramos. J: Kawakita. ER: Gnerra. CD: Issa.
JPM. (2014). Reduced bone and body mass in young male rats exposed to lead. BioMed Res Int 2014:
571065. http://dx.doi.org/10.1155/2014/571065.

Dearth. RK: Hinev. IK: Srivastava. V: Burdick. SB: Bratton. GR: Dees. WL. (2002). Effects of lead (Pb) exposure
during gestation and lactation on female pubertal development in the rat. Reprod Toxicol 16: 343-352.

http://dx.doi.org/ .1.0..1.0.1.6/S0890-6238(02)00037-0.

Dearth. RK: Hinev. IK: Srivastava. V: Les Dees. W: Bratton. GR. (2004). Low level lead (Pb) exposure during

gestation and lactation: Assessment of effects on pubertal development in Fisher 344 and Sprague-Dawley
female rats. Life Sci 74: 1139-1148. http://dx.doi.Org/.l.0.1016/ilfs.2003.07.033.

Dees. WL: Hinev. IK: Srivastava. VK. (2021). IGF-1 influences gonadotropin-releasing hormone regulation of
puberty [Review]. Neu roe ndocri no logy 111: 1151-1163. http://dx.doi.Org/.l.0.l.l.59/000514217.

Deierlein. AL: Teitelbaiim. SL: Windham. GC: Pinnev. SM: Gatvez. MP: Caldwell. KL: Jarrett. JM: Gajek. R:

Kushi	3Iff. MS. (2019). Lead exposure during childhood and subsequent anthropometry

through adolescence in girls. Environ Int 122: 310-315. http://dx.doi.Org/10.1016/i.envint.2018.ll.031.

Denhain. M: Schell. LM: Deane. G: Gatto. MY: Ravenscroft. J: DeCaprio. AP. (2005). Relationship of lead,

mercury, mirex, dichlorodiphenyldichloroethylene, hexachlorobenzene, and polychlorinated biphenyls to
timing of menarche among Akwesasne Mohawk girls. Pediatrics 115: el27-el34.

http://dx.doi.org/10.1542/peds.2004-l .1.6.1..

8-245


-------
Dev. S; Arinn. i; Das. M; Bhattachariee. CR; Dkhar. PS. (2001). Effect of prenatal lead toxicity on surface

ultrastructural features, elemental composition and infrared absorption characteristics of the skin of albino
mice. Cytobios 106(Suppl. 2): 245-254.

Dong. SY; Liang. DP; An. N; Jia. L; Shan. YJ; Chen. C; Sun. K; Nin. F; Li. HY; Fn. SB. (2009). The role of MAPK
and FAS death receptor pathways in testicular germ cell apoptosis induced by lead. Acta Biochim Biophys
Sin 41: 800-807. http://dx.doi.org/10.1093/abbs/gmp069.

Duan. Y; Peng. L; Shi. H; Jiang. Y. (20.1.7). The effects of lead on GAB Aergic interneurons in rodents. Toxicol Ind
Health 33: 867-875. http://dx.doi.org/10..1. .1.77/07482:	>902.

Dumitrescu. E; Alexandra. T; Sneiana. P. (2008a). Lead acetate impact on some markers of female reproductive
system performances (litter size, sex ratio) and physical development (vaginal opening) in rats. Bull Univ
Agric Sci Vet Med Cluj-Napoca Vet Med 65: 283-287. http://dx.doi.org/10.15835/buasvmcn-

vm:65:2:.1.554.

Dumitresc	rin. M. (2008b). The consequences of female rats chronic exposure to lead acetate in the

biomarkers emphasizing the hormonal disrupting potential of the reproductive function for in vivo
evaluation. Bull Univ Agric Sci Vet Med Cluj-Napoca Vet Med 65: 382-387.
http://dx.doi.Org/10.15835/bnasvmcn-vm:65:l:1241.

El Shafai. A; Zohdv. N: El Mulla. K; Hassan. M: Morad. N. (2011). Light and electron microscopic study of the
toxic effect of prolonged lead exposure on the seminiferous tubules of albino rats and the possible
protective effect of ascorbic acid. Food Chem Toxicol 49: 734-743.

http://dx.doi.org/10. .1.016/i.fct.201.0. .1. .1. .033.

Eum. KP; Weisskopf. MG: Nie. LH; Hn. H; Korrick. SA. (2014). Cumulative lead exposure and age at menopause
in the Nurses' Health Study cohort. Environ Health Perspect 122: 229-234.

http://dx.doi.org/10.1289/ehp. .1.206399.

Fleisch. AF: Burns. IS: Williams. PL: Lee. MM: Sergevev. O: Korrick. SA; Hauser. R. (2013). Blood lead levels
and serum insulin-like growth factor 1 concentrations in peripubertal boys. Environ Health Perspect 121:
854-858. http://dx.doi.Org/.l.0.1289/elip.1206105.

Flora. SJS; Tandon. SK. (.1.987). Influence of calcium disodium edetate on the toxic effects of lead administration in
pregnant rats. Indian J Physiol Pharmacol 31: 267-272.

Foster. WG; McMahon. A; Rice. DC. (.1.996). Subclinical changes in luteal function in cynomolgus monkeys with
moderate blood lead levels. J Appl Toxicol 16: 159-163. http://dx.doi.org/1.0..1.002/(SICI).1.099-
1263(.1.99603).1.6:2< .1.59: :AID-JAT326>3.0.CO;2-8.

Foster. WG; Stats. SI; McMahon. A. (.1.992). An ultrasound study of the effect of chronic lead exposure on
endometrial cycle changes in the female cynomolgus monkey. J Med Primatol 21: 353-356.

http://dx.doi.org/10. .1. .1. .1.1/i. .1.600-0684. .1.992. tb00603.x.

Franks. PA; Latighlin. NEC; Dierschke. DJ; Bowman. RE; Metier. PA. (1989). Effects of lead on luteal function in
rhesus monkeys. Biol Reprod 41: 1055-1062. http://dx.doi.org/10.1095/biotreprod41.6. .1.055.

Freire. C; A may a. E; Gil. F; Murcia. M; Hop. S; Casas. M; Vrijheid. M; Lertxundi. A; Irizar. A; Fernandez-Tardon.
G; Castro-Delgado. RV; Otea. N; Fernandez. MF. (2019). Placental metal concentrations and birth
outcomes: The Environment and Childhood (INMA) project. Int J Hyg Environ Health 222: 468-478.

http://dx.doi.org/10. .1.01.6/j. ijheh.201.8.1.2.0.1.4.

Gaiewska. K; Laskowska. M; Almeida. A; Pinto. E; Skorzvnska-Dziduszko. K; Blazewicz. A. (2021). Lead levels in
non-occupationally exposed women with preeclampsia. Molecules 26: 3051.
http://dx.doi.org/10.339Q/molecules26103Q51.

Garcia-Esauinas. E; Aragones. N; Fernandez. MA; Garcia-Sagredo. JM; de Leon. A; de Paz. C; Perez-Meixeira.
AM; Gil. E; Iriso. A; Cisneros. M; de Santos. A; Sanz. JC; Garcia.	nsio. A; Vioaue. J; Lopez-

Abe nte. G; Astray. J; Pollan. M; Martinez. M; Gonzalez. MJ; Perez-Gomez, B. (2014). Newborns and low
to moderate prenatal environmental lead exposure: Might fathers be the key? Environ Sci Pollut Res Int 21:
7886-7898. http://dx.doi.org/10.1007/sl 1356	38-6.

8-246


-------
Garcia-Esqiiin;	rez-Gomez. B; Fernandez-Navarro. P; Fernandez. MA; de Paz. C; Perez-Meixeira. AM; Gil

E; Iriso. A; Sanz. JC; Astray. J; Cisneros. M; de Santos. A; Asensi	rcia-Sagredo. JM; Garcia. JF;

Viotine. J; Lopez-Abente. G; Pottan. M; Gonzalez. MJ; Martinez, M; Aragones, N. (2013). Lead, mercury
and cadmium in umbilical cord blood and its association with parental epidemiological variables and birth
factors. BMC Public Health 13: 841. http://dx.doi.org/.1.0..1.186/1471-2458-13-841.

Gteason. KM; Valeri. L; Shankar. AH; Hasan. MPS. I; Ouamruzzaman. O: Rodrignes. EG; Christiani. DC: Wright.
RO; Bellinger. DC: Mazumdar. M. (2016). Stunting is associated with blood lead concentration among
Bangladeshi children aged 2-3 years. Environ Health 15: 103. http://dx.doi.org/	^12940-016-0190-

4.

Godinez-Solis. Y; de Jesua Solis-Heredia. M: Roa-Espitia. A; Parra-Forero	jnandez-Gonzalez. EO;

Hernandez-Ochoa. I: OuintaniHa-Vega. B. (2019). Low concentrations of lead decrease the sperm
fertilization ability by altering the acrosome reaction in mice. Toxicol Appl Pharmacol 380: 114694.
http://dx.doi.Org/10.1016/i.taap.2019.114694.

Gottenberg. AL: Hediger. ML: Lee. PA; Himes. JH: Buck Louis. GM. (2010). Association between lead and

cadmium and reproductive hormones in peripubertal U.S. girls. Environ Health Perspect 118: 1782-1787.

http://dx.doi.org/10.1289/ehp. 100.1.943.

Gotshan Iranponr. F; Kheiri. S. (20.1.6). Coadministration of calcium chloride with lead acetate can improve motility
of cauda epididymal spermatozoa in Swiss white mice. Int J Reprod Biomed 14: 141-144.

Gomes. M; Goncalves. A; Roeha. E; Sa. R; Alves. A; Sitva. J; Barros. A; Pereira. ML; Sonsa. M. (2015). Effect of
in vitro exposure to lead chloride on semen quality and sperm DNA fragmentation. Zygote 23: 384-393.
http://dx.doi.org/10.1017/S096719941300Q671.

Gomnla. A; Nowak-Szczepanska. N; Sebastian. A; Koziet. SM; Malina. RM; Ignasiak. Z. (2022). Age at menarche
in urban girls exposed to lead in the Copper Basin, Poland. Biology 11: 584.

http://dx.doi.org/10.3390/biology .1.1040584.

Goto. Y; Mandai. M; Na.kava.nK-:	iiazaki. S; Na.kava.ma. SF; Isobe. T; Sato. T; Nitta. H. (2021). Association of

prenatal maternal blood lead levels with birth outcomes in the Japan Environment and Children's Study
(JECS): A nationwide birth cohort study. Int J Epidemiol 50: 156-164.
http://dx.doi.org/10.1093/iie/dvaal62.

Govarts. E; Portengen. L; Lambreehts. N; Bruckers. L; Den Hond. E; Covaci. A; Nelen. V; Nawrot. TS; Loots. I;
Sioen. I; Baevens. W; Morrens. B; Schoeters. G; Vermeulen. R. (2020). Early-life exposure to multiple
persistent organic pollutants and metals and birth weight: Pooled analysis in four Flemish birth cohorts.
Environ Int 145: 106149. http://dx.doi.Org/10.1016/i.envint.2020.106149.

Govarts. E; Rernv. S; Backers. L; Den Hond. E; Sioen. I; Nelen. V; Baevens. W; Nawrot. TS; Loots. I; Van

Larebeke. N; Schoeters. G. (2016). Combined effects of prenatal exposures to environmental chemicals on
birth weight. Int J Environ Res Public Health 13: 495. http://dx.doi.org/10.3390/ijerphl3050495.

Graham. PL; Grace. CE; Braiin. AA; Schaefer. TL; Skelton. MR; Tang. PH; Vorhees. CV; Williams. MT. (2011).
Effects of developmental stress and lead (Pb) on corticosterone after chronic and acute stress, brain
monoamines, and blood Pb levels in rats. Int J Pev Neurosci 29: 45-55.
http://dx.doi.Org/10.1016/i.iidevneu.2010.09.008.

Gust in. K; Barman. M; Skroder. H; Jacobsson. B; Sandin. A; Sandberg. AS; Wold. AE; Vahter. M; Kippler. M.

(2021). Thyroid hormones in relation to toxic metal exposure in pregnancy, and potential interactions with
iodine and selenium. Environ Int 157: 106869. http://dx.doi.Org/.l.0.10.l.6/i.envint.202.L106869.

Gust in. K; Barman. M; Stravik. M; Levi. M; Englnnd-Qgge. L; Murray. F; Jacobsson. B; Sandberg. AS; Sandin. A;
Wold. AE; Vahter. M; Kippler. M. (2020). Low-level maternal exposure to cadmium, lead, and mercury
and birth outcomes in a Swedish prospective birth-cohort. Environ Pollut 265: 114986.
http://dx.doi.Org/10.1016/i.envpoL2020.114986.

Hasanein. P; Fazeli. F; Parviz. M; Roghani. M. (2018). Ferulic acid prevents lead-induced testicular oxidative stress
and suppressed spermatogenesis in rats. Andrologia 50: el2798. http://dx.doLo	fid..1.2798.

8-247


-------
Hassai	Sleweshv. M; Hassan. M; Noretdin. A. (2019). Thymoquinone attenuates testicular and

spermotoxicity following subchronic lead exposure in male rats: Possible mechanisms are involved. Life
Sci 230: 132-140. http://dx.doi.Org/10.1016/i.lfs.2019.05.067.

Hauser. R; Sergevev. O: Korrick. S: Lee. MM: Revich. B: Giti	rns. iS: Williams. PL. (2008). Association of

blood lead levels with onset of puberty in Russian boys. Environ Health Perspect 116: 976-980.

http://dx.doi.org/10.1289/ehp. 1.05.1.6.

H	i. 0: Chen. H: Weng. S: Luo. T: Zeng. X. (2016). Lead inhibits human sperm functions by reducing the

levels of intracellular calcium, cAMP, and tyrosine phosphorylation. Tohoku J Exp Med 238: 295-303.

littp ://dx. do i. o rg/ .1.0. .1.620/tj e m.23 8.295.

Hertin. M: Broberg. K: Igra. AM: Li. HQ: Harari. F: Vahter. M. (2019). Exploring telomere length in mother-

newborn pairs in relation to exposure to multiple toxic metals and potential modifying effects by nutritional
factors. BMC Med 17: 77. http://dx.doi.Org/.l.0.1186/sl.29.l.6-019-1309-6.

Hong. YC: Kulkarni. SS: Lim. YH: Kim. E: Ha. MN: Park. H: Kim. Y: Kim. BN: Chang. N: Oh. SY: Kim. YJ:

Park. C: Ha. EH. (2014). Postnatal growth following prenatal lead exposure and calcium intake. Pediatrics
134: 1151-1159. http://dx.doi.Org/10.1542/peds.2	8.

Hon. R: Hno. X: Zhang. S: Xn. C: Huang. Y: Xn. X. (2020). Elevated levels of lead exposure and impact on the

anti-inflammatory ability of oral sialic acids among preschool children in e-waste areas. Sci Total Environ
699: 134380. http://dx.doi.Org/.l.0.1016/i.scitotenv.201.9. .1.34380.

Hsu. PC: Chang. HY: Gno. YL: Lin. YC: Shih. TS. (2009). Effect of smoking on blood lead levels in workers and
role of reactive oxygen species in lead-induced sperm chromatinDNA damage. Fertil Steril 91: 1096-1103.
http://dx.doi.Org/10.1016/i.fertnstert.2008.01.005.

Hn. JMY: Arbuckle. TE: Janssei	iphear. BP: Zhuang. LH: Brann. JM: Chen. A: McCandless. LC. (2021).

Prenatal exposure to endocrine disrupting chemical mixtures and infant birth weight: A Bayesian analysis
using kernel machine regression. Environ Res 195: 110749.

http://dx.doi.org/10. .1.016/i.envres.202.1.. .1. .1.0749.

H	;ng. T: Cheng. Y: Holford. T: Lin. S: Leaderer. B: Qin. J: Bassig. BA: Shi. K: Zhang. Y: Niu. J: Zhn. Y:

t< i t .no. H: Che a l; iiang. J: Xn. S: Jin. Y. (2015). Distributions of heavy metals in maternal and cord
blood and the association with infant birth weight in China. J Reprod Med 60: 21-29.

Huang. BM; Lai. HY: Lin. MY. (2002). Concentration dependency in lead-inhibited steroidogenesis in MA-10
mouse Leydig tumor cells. J Toxicol Environ Health A 65: 557-567.

http://dx.doi.Org/.l.0.1080/15287390252808000.

Huang. BM: Liu. MY. (2004). Inhibitory actions of lead on steroidogenesis in MA-10 mouse Leydig tumor cells.
Arch Androl 50: 5-9. http://dx.doi.org/1.0..1.080/0.1.4850.1.0490250434.

Huang. LL: Huang. SF: Luo. XY: Li. LM: Chen. X: Zan. GH: Tan. YL: Lin. CO: Hon. OZ: Ge. XT: Chen. CF:
Hong. X: Tian. Y: Yang. AM: Chen. P: Li. Ml: Mo. ZN: Yang. XB. (2020). Associations of prenatal
exposure to multiple metals with testicular volume and anogenital distance in infant boys: A longitudinal
cohort study. Environ Int 143: 105900. http://dx.doi.Org/.l.0.1016/i.envint.2020.105900.

lavieoti. I: Carelti. G: Stanek. EI: Castellino. N: Li. Z: Calabrese. EX (2006). Low doses of dietary lead are

associated with a profound reduction in the time to the onset of puberty in female mice. Reprod Toxicol 22:
586-590. http://dx.doi.Org/10.1016/i.reprotox.2006.03.016.

lavieoti. I: Carelti. G: Stane*	steltino. N: Calabrese. EX (2004). Effects of low doses of dietary lead on

puberty onset in female mice. Reprod Toxicol 19: 35-41. http://dx.doi.Org/10.1016/i.reprotox.2004.06.013.

Ishitsuka. K: Y a ma mo t o - H anada. K: Yang. L: Mezawa. H: Konishi. M: Saito-Abe. M: Sasaki. H: Nishizato. M:

Sato. M: Koed	va. Y. (2020). Association between blood lead exposure and mental health in

pregnant women: Results from the Japan environment and children's study. Neurotoxicology 79: 191-199.

http://dx.doi.Org/10.1016/i.nenro.2020.06.003.

8-248


-------
Jan sen. EC; Zhou. L; Song. PXK; Sanchez. BN; Mercado. A; Hu. H; Solano. M; Peterson. KE; Tellez-Rojo. MM.

(2018). Prenatal lead exposure in relation to age at menarche: Results from a longitudinal study in Mexico
City. J Dev Orig Health Dis 9: 467-472. http://dx.doi.org/10..1.017/S20'	8000223.

Jedrvchowski.	rera. FP; Maiewska. R; Mrozek-Budzvn. D; Mroz. E; Roen. EL; Sowa. A; Jacek. R. (2015).

Depressed height gain of children associated with intrauterine exposure to polycyclic aromatic
hydrocarbons (PAH) and heavy metals: the cohort prospective study. Environ Res 136: 141-147.

http://dx.doi.Org/10.1016/i.envres.2014.08.047.

Jeltiffe-Pawtowski. LL; Miles. SO; Courtney. JG; Materna. 6; Charlton. V. (2006). Effect of magnitude and timing
of maternal pregnancy blood lead (Pb) levels on birth outcomes. J Perinatal 26: 154-162.

http://dx.doi.Org/10.1038/si.ip.7

Jia. X; Dong. T; Han. Y; Yue. Z; Zhang. P; Ren. J; Wang. Y; Wii. W; Yang. H; Giro. H; Zhang. GH; Cao. J. (2022).
Identifying the dose response relationship between seminal metal at low levels and semen quality using
restricted cubic spline function. Chemosphere 295: 133805.

http://dx.doi.Org/10.1016/i.chemosphere.2022..1.33805.

Jin. L; Zhang. L; Li. ZW; Liu. JM; Ye. RW; Ren. AG. (2013). Placental concentrations of mercury, lead, cadmium,
and arsenic and the risk of neural tube defects in a Chinese population. Reprod Toxicol 35: 25-31.
http://dx.doi.Org/10.1016/i.reprotox.2012.10.015.

Kagi. JHR; Vattee. BL. (.1.960). Metallothionein: A cadmium- and zinc-containing protein from equine renal cortex.
J Biol Chem 235: 3460-3465. http://dx.doi.org/10.1016/S0021-9258(18)64490-4.

Kahn. LG; Liu. XH; Rajovic. B; Popovac. D; Oberfield. S; Graziano. JH; Factor-Litvak. P. (2014). Blood lead

concentration and thyroid function during pregnancy: Results from the Yugoslavia Prospective Study of
Environmental Lead Exposure. Environ Health Perspect 122: 1134-1140.

http://dx.doi.org/10.1289/ehp. .1.307669.

Kasperezvl \ L isperczyk. S; Horak. S; Ostalowska. A; Grucka-Mamczar. E; Romuk. E; Olejel \ I'mfaier. E.
(2008). Assessment of semen function and lipid peroxidation among lead exposed men. Toxicol Appl
Pharmacol 228: 378-384. http://dx.doi.Org/10.1016/i.taap.2007..1.2.024.

Kerr. BT; Ochs-Balcom. HM; Lopez. P; Garcia-Vargas. GG; Rosado. JL; Cebrian. ME; Kordas. K. (2019). Effects
of ALAD genotype on the relationship between lead exposure and anthropometry in a Cohort of Mexican
children. Environ Res 170: 65-72. http://dx.doi.Org/10.1016/i.envres.2018..1.2.003.

Kim. C; Cathey. AL; Watkins. DJ; Mukheriee. B; Rosario-Pabon. ZY; Velez-Vega. CM; Alshawabkeh. AN;
Cordero. .IF; Meek . (2022). Maternal blood metal concentrations are associated with matrix
metalloproteinases (MMPs) among pregnant women in Puerto Rico. Environ Res 209: 112874.

http://dx.doi.org/10. .1.016/j.envres.2022. .1. .1.2874.

Kim. JH; Park. Y; Kim. SK; Moon. HB; Park. J; Choi. K; Kim. S. (2017). Timing of an accelerated body mass

increase in children exposed to lead in early life: A longitudinal study. Sci Total Environ 584-585: 72-77.

http://dx.doi.Org/10.1016/i.scitotenv.20.1.7.0.1. .1.22.

Kim. SS; Xu. XJ; Zhang. YL; Zheng. XB; Liu. RJ; Dietrich. KN; Reponen. T; Xie. CC; Sucharew. H; Huo. X;

Chen. AM. (2020). Birth outcomes associated with maternal exposure to metals from informal electronic
waste recycling in Guiyu, China. Environ Int 137: 105580. http://dx.doi.Org/.l.0.1016/i.envint.2020.105580.

Kresovich. JK; Argos. M; Tuiyk. ME. (20.1.5). Associations of lead and cadmium with sex hormones in adult males.
Environ Res 142: 25-33. http://dx.doi.Org/.l.0.1016/i.envres.2015.05.026.

Krieg. EF. Jr; Feng. HA. (20.1.1). The relationships between blood lead levels and serum follicle stimulating

hormone and luteinizing hormone in the National Health and Nutrition Examination Survey 1999-2002.
Reprod Toxicol 32: 277-285. http://dx.doi.org/10..1.01.6/i.reprotox.20.1. .1..05.0.1.2.

Knang. W; Chen. Z; Shi. K; Sun. H; Li. H; Huang. L; Bi. J. (2020). Adverse health effects of lead exposure on
physical growth, erythrocyte parameters and school performances for school-aged children in eastern
China. Environ Int 145: 106130. http://dx.doi.Org/.l.0.10.l.6/i.envint.2020.106130.

8-249


-------
Lai. GL; Yeh. CC: Yeh. CY; Chen. RY: Fu. CL; Chen. CH; Tzeng. CR. (2017). Decreased zinc and increased lead
blood levels are associated with endometriosis in Asian women. Reprod Toxicol 74: 77-84.
http://dx.doi.Org/10.1016/i.reprotox.2017.09.001.

Langhlin. NK; Bowman. RE: Franks. PA: Dierschke. DJ. (1987). Altered menstural cycles in rhesus monkeys
induced by lead. Toxicol Sci 9: 722-729. http://dx.doi.Org/10.1093/toxsci/9.4.722.

Leasnre. XL: Giddabasappa. A: Chanev. S: Johnson. .IE. J'r: Pothakos. K: Lau. YS: F	(2008). Low-level

human equivalent gestational lead exposure produces sex-specific motor and coordination abnormalities
and late-onset obesity in year-old mice. Environ Health Perspect 116: 355-361.

http://dx.doi.org/10.1289/ehp. .1.0862.

Lee. MS: Emu. KP: Go lam. M: Quanimzzaman. Q: Kile. ML: Mazmndar. M: Christiani. DC. (2021). Umbilical
cord blood metal mixtures and birth size in Bangladeshi children. Environ Health Perspect 129: 57006.

littp ://dx. do i. o rg/ .1.0.1289/.EHP7502.

Lee. S: Min. JY: Min. KB. (2020). Female infertility associated with blood lead and cadmium levels. Int J Environ
Res Public Health 17: 1794. http://dx.doi.org/10.3390/iierphl705.1.794.

Lee. TW: Kim. DH: Ryu. JY. (20.1.9). The effects of exposure to lead, cadmium and mercury on follicle-stimulating
hormone levels in men and postmenopausal women: Data from the Second Korean National Environmental
Health Survey (2012-2014). Ann Occup Environ Med 31: e21.

http://dx.doi.org/10.35371/aoem.2019.3Le21.

Leiva. KP: Rubio. J: Peratta. F: Gonzales. GF. (2011). Effect of Punica granatum (pomegranate) on sperm
production in male rats treated with lead acetate. Toxicol Mech Meth 21: 495-502.

http://dx.doi.org/10.3109/153765.1.6.20.1.1.555789.

Lewis. RC: Meekei	>15). Biomarkers of exposure to molybdenum and other metals in relation to testosterone

among men from the United States National Health and Nutrition Examination Survey 2011-2012. Fertil
Steril 103: 172-178. http://dx.doi.Org/.l.0.1016/i.fertnstert.2014.09.020.

Li. CJ: Yeh. CY: Chen. RY: Tzeng. CR: Han. BC: Chien. LC. (2015). Biomonitoring of blood heavy metals and
reproductive hormone level related to low semen quality. J Hazard Mater 300: 815-822.

littp ://dx. do i. o rg/ .1.0. .1.0.1.6/i.j haz ma t.20.1.5.08.027.

Li. D: Liang. C: Cao. Y: Zhu. D: Shen. L: Zhang. Z: Jiang. T: Zhang. Z: Zong. K: Lin. Y: Liang. D: Cao. Y: Ji. D:
Xn. X. (2022). The associations of serum metals concentrations with the intermediate and pregnancy
outcomes in women undergoing in vitro fertilization (IVF). Ecotoxicol Environ Saf 233: 113309.
http://dx.doi.Org/10.1016/i.ecoenv.2022.113309.

Li. J: Wang. H: Hao. JH: Chen. YH: Lin. L: Yn. Z: Fu. L: Tao. FB: Xn. DX. (2017a). Maternal serum lead level

during pregnancy is positively correlated with risk of preterm birth in a Chinese population. Environ Pollut
227: 484-489. http://dx.doi.Org/10.1016/i.envpol.2017.05.009.

Li. N: Qiao. M: Zhang. P: Li. X: Li. L. i: Yn. Z. (2016). The effects of early life lead exposure on the expression of
glycogen synthase kinase-3(3 and insulin-like growth factor 1 receptor in the hippocampus of mouse pups.
Biol Trace ElemRes 169: 114-120. http://dx.doi.org/10.1007/sl.20.1.1-015-0382-8.

Li. N: Zhao. G: Qiao. M: Shao. J: Li	H: Li. X: Yn. Z. (2014). The effects of early life lead exposure on the

expression of insulin-like growth factor 1 and 2 (IGF1, IGF2) in the hippocampus of mouse pups. Food
Chem Toxicol 63: 48-52. http://dx.doi.Org/10.1016/i.fct.201.3..1.0.037.

Li. S: Xn. J: Liu. Z: Yan. CH. (2017b). The non-linear association between low-level lead exposure and maternal
stress among pregnant women. Neurotoxicology 59: 191-196.

http://dx.doi.Org/.l.0.1016/i.neuro.2016.07.005.

Liao. K: Tsai. MS: Chang. CH: Chien. L: Ma	:ai. Y: Chen. M. (2015). Do the levels of maternal plasma trace

elements affect fetal nuchal translucency thickness? PLoS ONE 10: e0138145.

littp ://dx. do i. o rg/ .1.0.137.1. /j omnia Lpone .0138145.

8-250


-------
Lin. M; Yu. j; Su. Z; Sun. Y; Lin. Y; Xie. 0; Li. Z; Wang. L; Zhang. J; Jin. L; Ren. A. (2021). Associations
between prenatal exposure to cadmium and lead with neural tube defect risks are modified by single-
nucleotide polymorphisms of fetal MTHFR and SOD2: A case-control study. Environ Health 20: 66.

http://dx.doi.org/10.1186/sl2940-021-00752-9.

Liu. Y: Peterson. KE: Montgomery. K: Sanchez. BN: Zhang. ZZ: Afeiche. MC: Cantonwine. DE: Ettinger. AS:

Cantoral. A: Schnaas. L: Hu. H: Te'llez-Roio. MM. (2019a). Early lead exposure and childhood adiposity
in Mexico city. Int J Hyg Environ Health 222: 965-970. http://dx.doi.Org/.l.0.10.l.6/i.ijheh.2019.06.003.

Liu. Y: Teltez-Rojo. MM: Sanchez. BN: Zhang. Z: Afeiche. MC: Mercado-Garcia. A: Hu. H: Meeker. .I'D: Peterson.
KE. (2019b). Early lead exposure and pubertal development in a Mexico City population. Environ Int 125:
445-451. http://dx.doi.org/10. .1.01.6/i.envint.2019.02.02.1..

Liu. Z: He. CH: Chen. M: Yang. SH: Li. J: Lin. Y: Deng. Y: Li. N: Guo. YX: ^	XH. (2018). The effects of

lead and aluminum exposure on congenital heart disease and the mechanism of oxidative stress. Reprod
Toxicol 81: 93-98. http://dx.doi.org/10..1.01.6/i.reprotox.2018.07.08.1..

Logdberg. B: Berlin. M: Sehiitz. A. (.1.987). Effects of lead exposure on pregnancy outcome and the fetal brain of
squirrel monkeys. Scand J Work Environ Health 13: 135-145. http://dx.doi.org/10.5271/siweh.2069.

Lo pes. ACB. A: Peixe. TS: Mesas. AE: Paotielto. MMB. (2016). Lead exposure and oxidative stress: A systematic
review. In P de Voogt (Ed.), Reviews of environmental contamination and toxicology (Vol 236) (pp. 193-
238). Cham, Switzerland: Springer, http://dx.doi.org/10.1007/978-3-319-20013-2 3.

Louis. GM.B: Smarr. MM: Sundaram. R: Steuerwald. AJ: Sapra. KJ: Lu. ZH: Parsons. PJ. (2017). Low-level
environmental metals and metalloids and incident pregnancy loss. Reprod Toxicol 69: 68-74.

http://dx.doi.Org/10.1016/i.reprotox.	.0.1.1.

Louis. GMB: Sundaram. R: Schisterman. EF: Sweeney. AM: Lynch. CD: Gore-Langton. RE: Chen. Z: Kim. S:
Caldwell KL: Barr. DB. (2012). Heavy metals and couple fecundity, the LIFE Study. Chemosphere 87:
1201-1207. littp://dx.doi.org/.1.0. .1.0.1.6/j.chemosphere.20.1.2.0.1..0.1.7.

Mabrouk. A. (20.1.8). Therapeutic effect of thvmoquinone against lead-induced testicular histological damage in
male Wistar rats. Andrologia 50: el3014. http://dx.doi.org/10..1..1..1.1/and..1.30.1.4.

Madhavi. D: Devi. KR: Rao. KK: Reddv. PP. (2007). Modulating effect of Phyllanthus fruit extract against lead
genotoxicity in germ cells of mice. J Environ Biol 28: 115-117.

Massanvi. P: Lukac. N: Makarevich. AY: Chrene	rgacs. Z: Zakrzewski. M: Stawarz. R: Toman. R: Lazor. P:

Flesarova. S. (2007). Lead-induced alterations in rat kidneys and testes in vivo. J Environ Sci Health A Tox
Hazard Subst Environ Eng 42: 671-676. http://dx.doi.org/10.1080/10934520701244474.

Masso-Gonzal	Vntonio-Garcia. MT. (2009). Natural antioxidants protect against lead-induced damage during

pregnancy and lactation in rat's pups. Ecotoxicol Environ Saf 72: 2137-2142.

http://dx.doi.org/10. .1.016/i.ecoenv.2009.03.0.1.3.

Meeker. ID: Rossano. MG: Protas. B: Diamond. MP: Puscheck. E: Daly. D: Paneth. N: Wirth. JJ. (2008). Cadmium,
lead, and other metals in relation to semen quality: Human evidence for molybdenum as a male
reproductive toxicant. Environ Health Perspect 116: 1473-1479. http://dx.doi.org/10.1289/ehp. .1. .1.490.

Mendota. P: Brett. K: DiBari. IN: Pollack. AZ: Tandon. R: Shenassa. ED. (2013). Menopause and lead body burden
amongUS women aged 45-55, NHANES 1999-2010. Environ Res 121: 110-113.
http://dx.doi.Org/10.lQ16/i.envres.2012.12.Q09.

Mlkelson. CK: Troisi. J: LaLonde. A: Svnies. SJK: Thurston. SW: DiRe. LM: Adair. CD: Miller. RK: Richards.
SM. (2019). Placental concentrations of essential, toxic, and understudied metals and relationships with
birth outcomes in Chattanooga, TN. Environ Res 168: 118-129.

http://dx.doi.Org/10.1016/i.envres.2018.09.006.

8-251


-------
Mivashita. C; Saiio. Y; Ito. Y; Ikeda-Araki. A; Itoh. S; Yamazaki. K; Kobavashi. S; Ait Bamai. Y; Masuda. H;

Tamura. N; Itofa. M; Yamaguchi. T; Yamazaki. S; Kishi. R: Group. JEACsS. (2021). Association between
the concentrations of metallic elements in maternal blood during pregnancy and prevalence of abdominal
congenital malformations: The Japan Environment and Children's Study. Int J Environ Res Public Health
18: 10103. http://dx.doi.org/10.3390/iierphl81910103.

Moorman. WJ; Skaggs. SR; Clark. JC: Turner. TW; Sharpnack. DP: Murre	itnon. SD: Chapin. RE:

Schrader. SM. (1998). Male reproductive effects of lead, including species extrapolation for the rabbit
model. Reprod Toxicol 12: 333-346. http://dx.doi.org/10.1016/S0890-6238(98)00010-0.

Naderi. N: Souri. M: Esfahani. MHN: Haiian. M: Vash. NT. (2020). Ferulago angulata extract ameliorates
epididymal sperm toxicity in mice induced by lead and diazinon. Andrology 8: 706-718.

http://dx.doio:	sidr. .1.2730.

Naha. N: Chowdhurv. AR. (2006). Inorganic lead exposure in battery and paint factory: Effect on human sperm
structure and functional activity. J UOEH 28: 157-171. http://dx.doi.org/10.7888/iuoeh.28.157.

Naha. N: Manna. B. (2007). Mechanism of lead induced effects on human spermatozoa after occupational exposure.
Kathmandu Univ Med J 5: 85-94.

Naicker. N: Norris. SA: Matfaee. A: Becker. P: Richter. L. (2010). Lead exposure is associated with a delay in the
onset of puberty in South African adolescent females: Findings from the birth to twenty cohort. Sci Total
Environ 408: 4949-4954. http://dx.doi.Org/10.1016/i.scitotenv.2010.07.037.

Nampoothiri. LP: Gupta. S. (2006). Simultaneous effect of lead and cadmium on granulosa cells: A cellular model
for ovarian toxicity. Reprod Toxicol 21: 179-185. http://dx.doi.Org/10.1016/i.reprotox.2005.07.010.

Nampoothiri. LP: Gupta. S. (2008). Biochemical effects of gestational coexposure to lead and cadmium on
reproductive performance, placenta, and ovary. J Biochem Mol Toxicol 22: 337-344.

http://dx.doi.org/10.1002/ibt.20246.

Nava-Hernandez. MP: Hauad-Marroquin. LA: Bassol-Mavagoitia. S: Garcia-Arenas. G: Mercado-Hernandez. R:
Echavarri-Guzman. MA: Cerda-Flores. R.M. (2009). Lead-, cadmium-, and arsenic-induced DNA damage
in rat germinal cells. DNA Cell Biol 28: 241-248. lit!p://dx.do i. o rg/.1.0. .1.089/dna.2009.0860.

Nilsson. BO: Liung. L: Wide. M. (1.991). Electron microscopy and X-ray microanalyses of uterine epithelium from
lead-injected mice in an experimental delay of implantation. Arch Toxicol 65: 239-243.

http://dx.doi.org/10.1007/BF023073.1.5.

Nkomo. P: Richter. LM: Kagura. J: Mathee. A: Naicker. N: Norris. SA. (2018). Environmental lead exposure and
pubertal trajectory classes in South African adolescent males and females. Sci Total Environ 628-629:
1437-1445. littp://dx.doi.org/.1.0. .1.0.1.6/j.scitotenv.20.1.8.02..1.50.

Qguri. T: Ebara. T: Nakavama. SF: Sugiura-Qgasawara. M: Kamiiima. M: Japan Environment and Children's Study
Group. (2019). Association between maternal blood cadmium and lead concentrations and gestational
diabetes mellitus in the Japan Environment and Children's Study. Int Arch Occup Environ Health 92: 209-
217. http://dx.doi.org/10.1007/s00420-018-1367-7.

Otiveira. H: Spand. M: Santos. C: de Lourdes Pereira. M. (2009). Lead chloride affects sperm motility and acrosome
reaction in mice: Lead affects mice sperm motility and acrosome reaction. Cell Biol Toxicol 25: 341-353.

http://dx.doi.org/10.1007/sl.0565-008-9088-4.

Onunati. MM: Ahmadi. HN: Sabouri. S: Retana-Marquez. S: Abdoti. N: Rashno. S: Niknahad. H: Jamshidzadeh. A:
Mousavi. K: Rezaei. M: Akhlagh. A: Azarpira. N: Khoda	idari. R. (In Press) Glycine protects the

male reproductive system against lead toxicity via alleviating oxidative stress, preventing sperm
mitochondrial impairment, improving kinematics of sperm, and blunting the downregulation of enzymes
involved in the steroidogenesis. Environ Toxicol, http://dx.doi.org/10.1002/tox.23654.

Onunati. MM: Jamshidzadeh. A: Heidari. R: Sun. Z: Zaniiri. MJ: Khodaei. F: Mousapour. S: Ahmadi. F:

Javanmard. N: Yeganeh. BS. (2019). Carnosine and histidine supplementation blunt lead-induced
reproductive toxicity through antioxidative and mitochondria-dependent mechanisms. Biol Trace Elem Res
187: 151-162. http://dx.doi.org/10.1007/sl2011-Q18-1358-2.

8-252


-------
Osorio-Yanez. C; Sanchez-Guerra. M; Solano. M; Baccarelli. A; Wright. R; Sanders. AP; Tellez-Rojo. MM;
Tamavo-Ortiz. M. (2021). Metal exposure and bone remodeling during pregnancy: Results from the
PROGRESS cohort study. Environ Pollut 282: 116962. http://dx.doi.Org/10.1016/i.envpoL2021.116962.

Pant. N: Kumar. G: Upadhvav. AD: Patel. DK; Gupta. Y.K; Chaturvedi. PK. (2014). Reproductive toxicity of lead,
cadmium, and phthalate exposure in men. Environ Sci Pollut Res Int 21: 11066-11074.
http://dx.doi.org/10.1007/sll356-014-2986-5.

Pavlova. E; Ivanova. J; Pashkunova-Martic. I: Schaier. M: Petrova. E: Gluhcheva. Y; Dorkov. P: Helbich. TH:

Keppter. B; Koellensperger. G. (2021). Ameliorative effects of deferiprone and tetraethylammonium salt of
salinomycinic acid on lead-induced toxicity in mouse testes. Environ Sci Pollut Res Int 28: 6784-6795.
http://dx.doi.org/10.1007/sll356-020-1096Q-4.

Pi. X: Qiao. YR: Wei. YH; Jin. L; Li. ZW; Lin. J.F; Zhang. YL; Wang. LL; Lin. YO; Xie. O; Ren. AG. (2018).

Concentrations of selected heavy metals in placental tissues and risk for neonatal orofacial clefts. Environ
Pollut 242: 1652-1658. http://dx.doi.Org/10.1016/i.envpol.2018.07.112.

Piao. FY: Cheng. FY: Chen. HB; Li. G: Sun. X; Lin. S: Yamauchi. T; Yokoyama. K. (2007). Effects of zinc
coadministration on lead toxicities in rats. Ind Health 45: 546-551.
http://dx.doi.org/10.2486/indhealth.45.546.

Piasek. M; Kostiat. K. (.1.991). Reversibility of the effects of lead on the reproductive performance of female rats.
Reprod Toxicol 5: 45-51. http://dx.doi.org/.1.0..1.016/0890-6238(91)90109-5.

Pittai. P; Pandva. C: Bhatt. N: Gupta. SS. (2012). Biochemical and reproductive effects of gestational/lactational
exposure to lead and cadmium with respect to testicular steroidogenesis, antioxidant system, endogenous
sex steroid and cauda-epididymal functions. Andrologia 44: 92-101. http://dx.doi.oi	1.439-

0272.2010.01109.x.

Pittai. P: Pandva. C: Gupta. S: Gupta. S. (20.1.0). Biochemical and molecular effects of gestational and lactational
coexposure to lead and cadmium on ovarian steroidogenesis are associated with oxidative stress in F1
generation rats. J Biochem Mol Toxicol 24: 384-394. http://dx.doi.org/10.1002/ibt.20351.

Pine. MP; Hinev. IK; Dearth. RK; Bration. GR; Dees. WL. (2006). IGF-1 administration to prepubertal female rats
can overcome delayed puberty caused by maternal Pb exposure. Reprod Toxicol 21: 104-109.
http://dx.doi.Org/10.1016/i.reprotox.2005.07.003.

Pinon-Lataittade. G; Thoreux-Manlav. A; Coffignv. H; Masse. R; Sonfir. JC. (1995). Reproductive toxicity of
chronic lead exposure in male and female mice. Hum Exp Toxicol 14: 872-878.
http://dx.doi.oi	>2719501401103.

Pollack. AZ; Schisterman. EF; Goldman. LR; Mnmford. SL; Albert. PS; Jones. RL; Wactawski-Wende. i. (2011).
Cadmium, lead, and mercury in relation to reproductive hormones and anovulation in premenopausal
women. Environ Health Perspect 119: 1156-1161. http://dx.doi.org/10.1289/ehp. .1.003284.

Rabito. FA; Koeak. M; Werthmann. DW; Tvlavsl	3almer. CP; Parsons. PJ. (2014). Changes in low levels of

lead over the course of pregnancy and the association with birth outcomes. Reprod Toxicol 50: 138-144.

http://dx.doi.Org/10.1016/i.reprotox.	006.

Rahman. ML; Oken. E; Hivert. MF; Rifas-Shiman. S; Lin. PIP; Cotieino. E; Wright. RO; Amarasiriwardena. C;

Henn. BGC; Gold. PR; Coull. BA; Cardenas. A. (2021). Early pregnancy exposure to metal mixture and
birth outcomes: A prospective study in Project Viva. Environ Int 156: 106714.
http://dx.doi.Org/10.lQ16/i.envint.2021.lQ6714.

Raihan. Ml; Briskin. E; Mahfiiz. M; Islam. MM; Mondal. P; Hossain. MI; Ahmed. AM'S; Hague. R; Ahmed. T.

(2018). Examining the relationship between blood lead level and stunting, wasting and underweight—A
cross-sectional study of children under 2 years-of-age in a Bangladeshi slum. PLoS ONE 13: e0197856.

http://dx.doi.org/10.1371/ionnial.pone.01.97856.

Rao Barknr. R; Baity. LK. (20.1.6). Comparison of the developmental milestones and preweaning neurobehavioral
parameters in rat pups exposed to lead (Pb) during gestation, lactation and pregestation period. Drug Chem
Toxicol 39: 248-255. http://dx.doi.org/.1.0.3.1.09/0.1.480545.20.1.5..1.082.1.36.

8-253


-------
Renzetti. S; Just. AC: Bums. HH; Oken. E; Amarasiriwardena. C; Svensson. K; Mercado-Garcia. A; Cantoral A;
Schnaas. L; Baccarelli. AA; Wright. RO; Tellez-Rojo. MM. (2017). The association of lead exposure
during pregnancy and childhood anthropometry in the Mexican PROGRESS cohort. Environ Res 152: 226-
232. http://dx.doi.Org/10.10.l.6/i.envres.20.l.6.10.014.

Rodosthenous. RS: Bums. HH: Svensson. K: Amarasiriwardena. CI: Cantoral. A: Schnaas. L: Mercado-Garcia. A:
Coull BA: Wright. RO: Tellez-Rojo. MM: Baccarelli. AA. (2017). Prenatal lead exposure and fetal
growth: Smaller infants have heightened susceptibility. Environ Int 99: 228-233.

http://dx.doi.org/10. .1.0.1.6/i.envint.20.1.6. .1.1.023.

Ronis. MJJ: Aronson. J: Gao. GG: Hogue. W: Skinner. RA; Badger. TM: Lumpkin. CK. jr. (2001). Skeletal effects
of developmental lead exposure in rats. Toxicol Sci 62: 321-329. http://dx.doi.Org/10.1093/toxsci/62.2.321.

Ronis. MJJ: Badger. TM': Shema. SJ: Roberson. PK: Shaikh. F. (1996). Reproductive toxicity and growth effects in
rats exposed to lead at different periods during development. Toxicol Appl Pharmacol 136: 361-371.

http://dx.doi.org/10.1006/taap.1996.0044.

Ronis. MJJ: Badger. TM: Shema. SJ: Roberson. PK: Shaikh. F. (1998a). Effects on pubertal growth and

reproduction in rats exposed to lead perinatally or continuously throughout development. J Toxicol Environ
Health A 53: 327-341. http://dx.doi.org/10. .1.080/00984.1.098.1.593.1.2.

Ronis. MJJ: Badger. TM: Shema. SJ: Roberson. PK: Tempter. L: Ringer. D: Thomas. PE. (1998b). Endocrine

mechanisms underlying the growth effects of developmental lead exposure in the rat. J Toxicol Environ
Health A 54: 101-120. http://dx.doi.org/10.1080/009841098158944.

Ronis. MJJ: Candy. J: Badger. T. (1998c). Endocrine mechanisms underlying reproductive toxicity in the
developing rat chronically exposed to dietary lead. J Toxicol Environ Health A 54: 77-99.

http://dx.doi.org/10.1080/009841098158935.

Rubio. J: Riqueros. MI: Gasco. M: Yucra. S: Miranda. S: Gonzales. GF. (2006). Lepidium meyenii (Maca) reversed
the lead acetate induced—Damage on reproductive function in male rats. Food Chem Toxicol 44: 1114-
1122. http://dx.doi.Org/10.1016/i.fct.2006.01.007.

Sainath. SB: Meena. R: Supriva. C. h: Reddv. KP: Reddv. PS. (2011). Protective role of Centella asiatica on lead-
induced oxidative stress and suppressed reproductive health in male rats. Environ Toxicol Pharmacol 32:
146-154. http://dx.doi.org/10. .1.016/i.etap.201.1..04.005.

Satawu. EO: Adeevo. OA: Falokun. OP: Yusuf. UA; Oyerinde. A: Adeleke. AA. (2009). Tomato (Lycopersicon
esculentum) prevents lead-induced testicular toxicity. Journal of Human Reproductive Sciences 2: 30-34.

http://dx.doi.org/10.4103/0974-1208.51346.

Sateh. HA: Abd El-Aziz. GS: Mustafa, HN: El-Fark. M: Mat A: Aburas. M: Deifalla. AH. (2019). Thymoquinone
ameliorates oxidative damage and histopathological changes of developing brain neurotoxicity. J
Histotechnol 42: 116-127. http://dx.doi.org/10.1080/01478885.2Q19.1619654.

Sateh. HA: Abdel El-Aziz. GS: Mustafa.. HN: Sal	c Mat. AO: Deifatta. AHS: Aburas. M. (2018). Protective

effect of garlic extract against maternal and foetal cerebellar damage induced by lead administration during
pregnancy in rats. Folia Morphol (Warsz) 77: 1-15. http://dx.doi.org/10.5603/FM.a2017.0063.

Sanchez-Guerra. M: Peng. C: Trevisi. L: Cardenas. A: Wilson. A: Osorio-Yanez. C: Niedzwiecki. MM: Zhong. J:
Svensson. K: Acevedo. MT: Sotano-Gonzatez. M: Amarasiriwardena. CJ: Estrada-Gutierrez. G: Brennan.
KJM: Schnaas. L: Just. AC: Lane. HE: Wright. RJ: Tellez-Rojo, MM;... Baccarelli, AA. (2019). Altered
cord blood mitochondrial DNA content and pregnancy lead exposure in the PROGRESS cohort. Environ
Int 125: 437-444. http://dx.doi.Org/.l.0.10.l.6/i.envint.2019.01.077.

Sanders. AP: Burris. HH: Just. AC: Motta. V: Amarasiriwardena. C: Svensson. K: Oken. E: Sotano-Gonzatez. M:
Mercado-Garcia. A; Pantic. I; Schwartz. J; Tellez-Rojo. MM; Baccarelli. AA; Wright. RO. (2015). Altered
miRNA expression in the cervix during pregnancy associated with lead and mercury exposure.

Epigenomics 7: 885-896. http://dx.doi.org/10.2217/epi.15.54.

8-254


-------
Santos. S; Voerman. E; Amiano. P; Barros. H; Beilin. LI; Bergstrom. A; Charles. MA; Chatzi. L; Chevrier. C;
Chrousos. GP; Corpeteiin. E; Costa. O; Costet. N; Crozier. S; Devereux. G; Dovon. M; Eggesbe. M;
Fantinl. MP; Farefai. S; ... Jaddoe. V. (2019). Impact of maternal body mass index and gestational weight
gain on pregnancy complications: An individual participant data meta-analysis of European, North
American and Australian cohorts. BJOG 126: 984-995. http://dx.doi.oi	3528.15661.

Satapathv. S; Panda. CR. (20.1.7). Toxic metal ion in seafood: Meta-analysis of human carcinogenic and non-

carcinogenic threat assessment, a geomedical study from Dhamra and Puri, Odisha. Hum Ecol Risk Assess
23: 864-878. http://dx.doi.org/10.1080/10807039.2017.1288563.

Schneider. IS; Anderson. DW: Kidd. SK; Sobotewski. M; Corv-Slecht; . (2016). Sex-dependent effects of lead
and prenatal stress on post-translational histone modifications in frontal cortex and hippocampus in the
early postnatal brain. Neurotoxicology 54: 65-71. http://dx.doi.org/10. .1.016/i.neuro.201.6.03.0.1.6.

Selevan. SG; Rice. DC; Hogan. L \ ! nling. SY; Pfahles-Hutchenp .\ Kahel. .1. (2003). Blood lead concentration
and delayed puberty in girls. N Engl J Med 348: 1527-1536. http://dx.doi.org/10.1056/NEJMoa020880.

Shan. G; Tang. T; Zhang. XB. (2009). The protective effect of ascorbic acid and thiamine supplementation against
damage caused by lead in the testes of mice. J Huazhong Univ Sci Technolog Med Sci 29: 68-72.

http://dx.doi.org/10.1007/sl .1.596-009-0.1.14-4.

Shapiro. GD; Dodds. L; Arbuckle. TE; Ashley-Mart in. J; Eraser. W; Fisher. M; Taback. S; Keely. E; Bouchard. MF;
Monnier. P; Daltaire. R; Morisset. AS; Ettinger. AS. (2015). Exposure to phthalates, bisphenol A and
metals in pregnancy and the association with impaired glucose tolerance and gestational diabetes mellitus:
The MIREC study. Environ Int 83: 63-71. http://dx.doi.Org/10.1016/i.envint.2015.05.016.

SI	an. CPS; Man. GKY; Chan. DY.L; Wong. M.H; Li. TC. (2021). Associations between blood metal/

metalloid concentration and human semen quality and sperm function: A cross-sectional study in Hong
Kong. J Trace Elem Med Biol 65: 126735. http://dx.doi.org/10..1.0.1.6/i jtenib.202.1..126735.

SI	Chen. HY; Christensen. K; Handler. A; Tuiyk. M.E; Argos. M. (2021). Prenatal exposure to multiple

metals and birth outcomes: An observational study within the National Children's Study cohort. Environ Int
147: 106373. http://dx.doi.Org/10.1016/i.envint.2020.106373.

Signes-Pastor. AJ; Desai. G; Garcia-ViHarino. M; Karagas. MR; Kordas. K. (2021). Exposure to a mixture of metals
and growth indicators in 6-11-year-old children from the 2013-2016 NHANES. Exposure and Health 13:
173-184. http://dx.doi.org/10.1007/sl.2403-020-00371-8.

Singh. C; Saxena. DK; Murthv. RC; Chandra. SY. (1993). Embryo-fetal development influenced by lead exposure
in iron-deficient rats. Hum Exp Toxicol 12: 25-28. http://dx.doi.org/10.1177/0960327193012001Q5.

Sitarik. AR; Arora. M; Austin. C; Bielak. LF; Eggers. S; Johnson. CC; Lynch. SY; Park. SK; Wii. KHH; Yong.

GJM; Cassidv-Bushrow. AE. (2020). Fetal and early postnatal lead exposure measured in teeth associates
with infant gut microbiota. Environ Int 144: 106062. http://dx.doi.org/10. .1.016/i.envint.2020. .1.06062.

Stawinska. T; Ignasiak. Z; Little. BB; Malina. RM. (2012). Short-term secular variation in menarche and blood lead
concentration in school girls in the copper basin of southwestern Poland: 1995 and 2007. Am J Hum Biol
24: 587-594. http://dx.doi.org/10.1002/aihb.22272.

Sobotewski. M; Abston. K; Conrad. K; Marvin. E; Harvey. K; Susiario. M; Corv-Slecht; . (2020). Lineage- and
sex-dependent behavioral and biochemical transgenerational consequences of developmental exposure to
lead, prenatal stress, and combined lead and prenatal stress in mice. Environ Health Perspect 128: 27001.
http://dx.doi.org/10.1289/EHP4977.

Sokol. RZ; Berman. N. (1.991). The effect of age of exposure on lead-induced testicular toxicity. Toxicology 69:
269-278. http://dx.doi.org/10..1.016/03Q0-483X(91)90.1.86-5.

Sokol RZ; Madding. CE; Swerdloff. RS. (.1.985). Lead toxicity and the hypothalamic-pituitary-testicular axis. Biol
Reprod 33: 722-728. http://dx.doi.Org/10.1095/biolreprod33.3.722.

Sokol. RZ; Oku da. H; Nagter. HM; Berman. N. (1994). Lead exposure in vivo alters the fertility potential of sperm
in vitro. Toxicol Appl Pharmacol 124: 310-316. http://dx.doi.org/10.1006/taap. 1.994. .1.037.

8-255


-------
Soomro. MH; Baiz. N; Huei G; Yazbeck. C; Botton. j; Heude. 6; Bornehag. CG; Annesi-Maesano. I; EDEN
Mother-Child Cohort Study Group. (2019). Exposure to heavy metals during pregnancy related to
gestational diabetes mellitus in diabetes-free mothers. Sci Total Environ 656: 870-876.
http://dx.doi.Org/10.1016/i.scitotenv.2018.ll.422.

Srivastava. V: Dearth. RK; Hinev. JK: Ramirez. LM: B ration. GR: Dees. W. (2004). The effects of low-level Pb on
steroidogenic acute regulatory protein (StAR) in the prepubertal rat ovary. Toxicol Sci 77: 35-40.

http://dx.doi.Org/.l.0.1093/toxsci/kfg249.

Sukhn. C: Awwad. J: Ghantous. A: Zaatari. G. (2018). Associations of semen quality with non-essential heavy

metals in blood and seminal fluid: Data from the Environment and Male Infertility (EMI) study in Lebanon.
J Assist Reprod Genet 35: 1691-1701. http://dx.doi.org/10.1007/slQ815-018-1236-z.

Takeuchi. M: Yoshida. S: Kawakami. C; Kawakami. K: Ito. S: Japan Environment and Children's Study Group.

(2022). Association of maternal heavy metal exposure during pregnancy with isolated cleft lip and palate in
offspring: Japan Environment and Children's Study (JECS) cohort study. PLoS ONE 17: e0265648.

http://dx.doi.Org/.l.0.1371/ioumaLpone.0265648.

Tamavo v Ortiz. M: Tellez-Roio. MM: Wright. RJ: Coull. BA: Wright. RO. (2016). Longitudinal associations of age
and prenatal lead exposure on Cortisol secretion of 12-24 month-old infants from Mexico City. Environ
Health 15: 41. http://dx.doi.org/	?s.l.2940-0.1.6-01.24-1.

Tang. ML: Xu. CY: Lin. N: Liu. K: Zhai	fti. XW: Liu. WP. (2016). Lead, mercury, and cadmium in

umbilical cord serum and birth outcomes in Chinese fish consumers. Chemosphere 148: 270-275.

http://dx.doi.org/10. .1.016/j.chemosphere.201.6.0.1. .058.

Tapisso. JT: Marques. CC: Mathias. ML: Ramalhinho. MG. (2009). Induction of micronuclei and sister chromatid
exchange in bone-marrow cells and abnormalities in sperm of Algerian mice (Mus spretus) exposed to
cadmium, lead and zinc. Mutat Res Genet Toxicol Environ Mutagen 678: 59-64.

http://dx.doi.org/10. .1.01.6/j. mrgentox.2009.07.001.

Tartagtione. AM: Serafini. MM: Raggi. A: lacoponi. F: Zianni. E: Scatfari. A: Minghetti. L: Ricceri. L: Cubadda. F:
Calamandrei. G: Viviani. B. (2020). Sex-dependent effects of developmental lead exposure in Wistar rats:
Evidence from behavioral and molecular correlates. International Journal of Molecular Sciences 21: 2664.
http ://dx. do i .o rg/ .1.0.33 90/ij ms2.1.082664.

Tatsuta. N: Iwai-Shimada. M: Nakavama. SF: Iwama. N: Metoki. H: Arima. T: Sakural. K: Anal. A: Asato. K:
Kurivama. S: Sugawara. J: Suzuki. K: Yaegashi. N: Kamiiima. M: Nakai. K: Group. JEaCsS. (2022a).
Association between whole blood metallic elements concentrations and gestational diabetes mellitus in
Japanese women: The Japan Environment and Children's Study. Environ Res 212: 113231.

http://dx.doi.org/10.1016/i.envres.2022. .1. .1.3231.

Tatsuta. N: Kurokawa. N: Nakai. K: Suzuki. K: Iwai-Shimada. M: Murata. K: Satoh. H. (2017). Effects of

intrauterine exposures to polychlorinated biphenyls, methylmercury, and lead on birth weight in Japanese
male and female newborns. Environ Health Prev Med 22: 39. http://dx.doi.org/10.1186/sl2199-Q17-Q635-
6.

Tatsuta. N: Nakai. K: Nakavama. SF: Takeuchi. A: Arima	gashi. N: Kamiiima. M: Japan Environment and

Children's Study Group: Yamazaki. S: Olrva. Y: Kishi. R: Hashimoto. K: Mori. C: Ito. S: Yamagata. Z:
Inadera. H: Nakavama. T: Iso. H: Shi ma. M:... Katoh, T. (2022b). Effects of maternal exposure to lead on
secondary sex ratio in Japan: The Japan Environment and Children's Study. Sci Total Environ 817: 152726.
http://dx.doi.Org/lQ.1016/i.scitotenv.2Q21.152726.

Taupeau. C: Poupon. J: Treton. D: Brosse. A: Richard. Y: Machelon. V. (2003). Lead reduces messenger RNA and
protein levels of cytochrome P450 aromatase and estrogen receptor (3 in human ovarian granulosa cells.

Biol Reprod 68: 1982-1988. http://dx.doi.org/10.1095/biolreprod. 102.009894.

Taylor. CM: Golding. J: Emond. AM. (20.1.4). Girl or boy? Prenatal lead, cadmium and mercury exposure and the
secondary sex ratio in the ALSPAC study. Reprod Toxicol 46: 137-140.

http://dx.doi.Org/10.1016/i.reprotox.	0.1.1.

8-256


-------
Taylor. CM; Gotding. i; Etnoncl. AM. (20.1.5). Adverse effects of maternal lead levels on birth outcomes in the
ALSPAC study: A prospective birth cohort study. BJOG 122: 322-328. http://dx.doi.ort
0528.12756.

Taylor. CM: Tilling. K; Gotding. j; Emond. AM. (2016). Low level lead exposure and pregnancy outcomes in an
observational birth cohort study: Dose-response relationships. BMC Research Notes 9: 291.

http://dx.doi.org/10. .1.1.86/s .1.3.1.04-0.1.6-2092-5.

Teiion. C: Otmo. R; Blanco. D; Romero. A; Teiion. J'M. (2006). Low doses of lead: Effects on reproduction and
development in rats. Biol Trace ElemRes 111: 151-165. fattp://dx.doi.org/.1.0..1.385/.B1TE

Telisman. S: Colak. B; Pizent. A; Jurasovic. J; Cvitkovic. P. (2007). Reproductive toxicity of low-level lead
exposure in men. Environ Res 105: 256-266. http://dx.doi.Org/10.1016/i.envres.2007.05.011.

Thomas. S: Arbuckle. TE; Fisher. M: Eraser. WD: Ettinger. A; King. W. (2015). Metals exposure and risk of small-
for-gestational age birth in a Canadian birth cohort: The MIREC study. Environ Res 140: 430-439.

http://dx.doi.Org/10.1016/i.envres.20.1.5.04.0.1.8.

Thorenx-Mant	aseogne. C; Segretain. D; Jegou. B; Pinon-Lataillade. G. (1995). Lead affects

steroidogenesis in rat Leydig cells in vivo and in vitro. Toxicology 103: 53-62.

http://dx.doi.org/ .1.0..1.0.1.6/0300-483Xf95)03.1

Tian. T; Yin. S; Jin. L; Lin. J; Wang. C; Wei. J; Lin. M; Li. Z; Wang. L; Yin. C; Ren. A. (2021). Single and mixed
effects of metallic elements in maternal serum during pregnancy on risk for fetal neural tube defects: A
Bayesian kernel regression approach. Environ Pollut 285: 117203.

http://dx.doi.org/10. .1.016/i.envpoL202.1.. .1. .1.7203.

Totnnav. HE; Siikiir. YE; Ozkavukeu. S; Seval. MM; Ates. C; Tiirksov. VA; Eeemi	bekogtu. CS; Ozmen. B;

Berker. B; Sonmezer. M. (2016). Heavy metal and trace element concentrations in blood and follicular
fluid affect ART outcome. Eur J Obstet Gynecol Reprod Biol 198: 73-77.

http://dx.doi.org/10. .1.016/i.eiogrb.201.6.0.1. .00.1..

Tornomn. HY; Mostafa. GA; Ismail. NA; Ahmed. SM. (2010). Lead exposure and its association with pubertal
development in school-age Egyptian children: Pilot study. Pediatrics International 52: 89-93.
http://dx.doi.o:	1442-200X.2009.02893.X.

Tsuii. M; Shibata. E; Askew. DJ; Morokuma. S; Aiko. Y; Senin. A; Araki. S; Sanefuii. M; Ishihara. Y; Tanaka. R;
Kusuhara. K; Kawamoto. T. (2019). Associations between metal concentrations in whole blood and
placenta previa and placenta accreta: the Japan Environment and Children's Study (JECS). Environ Health
Prev Med 24: 40. http://dx.doi.org/10.1186/sl2199-019-0795-7.

Tsuii. M; Shibata. E; Morokuma. S; Tanaka. R; Senin. A; Araki. S; Sanefuii. M; Korivama. C; Yamamoto. M;

Ishihara. Y; Kusuhara. K; Kawamot	to. H; Kishi. R; Yaegashi. N; Hashimoto. K; Mori. C; Ito. S;

Yamagata. Z; Inadera. H; Kamiiima. M; Nakavama. T; Iso, H; Shima, M; Hirooka, Y; Suganuma, N;
Kusuhara, K; Katoh, T. (2018). The association between whole blood concentrations of heavy metals in
pregnant women and premature births: The Japan Environment and Children's Study (JECS). Environ Res
166: 562-569. http://dx.doi.Org/10.lQ16/i.envres.2018.Q6.025.

U.S. EPA. (2006). Air quality criteria for lead [EPA Report]. (EPA/600/R-05/144aF-bF). Research Triangle

Park, NC. http://cfpub.epa.gov/ncea/CFM/recordisplay.cfm?deid=158823.

U.S. EPA. (20.1.3). Integrated science assessment for lead [EPA Report]. (EPA/600/R-10/075F). Washington, DC.
https://nepis.epa.gov/Exe/ZyPURL.cgi?Dockev=P100K82L.txt.

U.S. EPA. (20.1.5). Preamble to the Integrated Science Assessments [EPA Report]. (EPA/600/R-15/067).

Research Triangle Park, NC: U.S. Environmental Protection Agency, Office of Research and Development,
National Center for Environmental Assessment, RTP Division, https://cfpnb.epa.gov/ncea/isa/
recordisplay.cfm?deid=3.1.0244.

Udefa. AL; Amama. EA; Archibong. EA; Nwangwa. JN; Ada ma. S; Invang. VU; Invaka. GU; Ain. GJ; Okpa. S;
Inah. IP. (2020). Antioxidant, anti-inflammatory and anti-apoptotic effects of hydro-ethanolic extract of
Cyperus esculentus L. (tigernut) on lead acetate-induced testicular dysfunction in Wistar rats. Biomed
Pharmacother 129: 110491. http://dx.doi.org/10..1.01.6/i.biopha.2020. .1..1.049.1..

8-257


-------
Vigeh. M; Yokoyama. K; Sevedaghamiri. Z; Shinohara. A; Matsukawa. T; Chiba. M; Yunesian. M. (2011). Blood
lead at currently acceptable levels may cause preterm labour. Occup Environ Med 68: 231-234.

http://dx.doi.Org/.l.0.1136/oem.2009.050419.

Vigeh. M: Yunesian. M: Matsukawa. T: Shamsipour. M: Jeddi. MZ: Rastkari. N: Hassanvand. MS: Shariat. M:
Kashani. H: Pirjanl. R: Effatpanah. M: Shirazi. M: Shariatpanahi. G: Ohtani. K: Yokoyama. K. (2021).
Prenatal blood levels of some toxic metals and the risk of spontaneous abortion. J Environ Health Sci Eng
19: 357-363. http://dx.doi.org/10.1007/s40201-020-006Q8-3.

Wang. CH: Zhang. Y: Liang. JC: Shan. G: Wang. Y: Shi. O. (2006). Impacts of ascorbic acid and thiamine
supplementation at different concentrations on lead toxicity in testis. Clin Chim Acta 370: 82-88.

http://dx.doi.org/10. .1.016/i.cca.2006.0.1. .023.

Wang. H: Ji. YL: Wang. O: Zhao. XF: Ning. H: Lin. P: Zhang. C: Yn. T: Zhang. Y: Meng. >	. (2013a).

Maternal lead exposure during lactation persistently impairs testicular development and steroidogenesis in
male offspring. J Appl Toxicol 33: 1384-1394. http://dx.doi.org/10.1002/iat.2795.

Wang. H: Li. J: Hao. JH: Chen. YH: Lin. L: Yu. Z: Fu. L: Tao. FB: Xn. DX. (2017a). High serum lead concentration
in the first trimester is associated with an elevated risk of small-for-gestational-age infants. Toxicol Appl
Pharmacol 332: 75-80. http://dx.doi.Org/.l.0.1016/i.taap.2017.07.020.

Wang. J: Gao. ZY; Yan. J: Ying. XL: Tong. SL: Yan. CH. (2017b). Sex differences in the effects of prenatal lead
exposure on birth outcomes. Environ Pollut 225: 193-200. http://dx.doi.org/10. .1.01.6/i .envpoL2017.03.03.1..

Wang. Y: Fant	tang. SX: Chen. L: Fan. GO: Wang. CH. (2013b). The chronic effects of low lead level on the

expressions of Nr£2 andMrpl of the testes in the rats. Environ Toxicol Pharmacol 35: 109-116.

http://dx.doi.org/10. .1.016/i.etap.201.2.1.2.00.1..

Wang. Y: Zhai	;en. X: Wu. WW: Feng. YL: Yang. HL: Li. M: Xie. BJ: Guo. PG: Warren. JL: Shi. XM:

Wang. SP: Zhang. YW. (2019). Multiple metal concentrations and gestational diabetes mellitus in Taiyuan,
China. Chemosphere 237: 124412. http://dx.doi.org/10. .1.01.6/j.ehemosphere.2019. .1.244.1.2.

Wang. YY: Hu. HY: Li. H: Ma. HY: Xn. FS: On. BM. (2014). Effects of lead exposure on placental cellular
apoptosis and endoplasmic reticulum stress in rats. Chin Med J 127: 1744-1748.

http://dx.doi.Org/10.3760/cma.j.issn.0366-6999.20132924.

Wang. YY: Sni. KX: Hong. LI: Ma. HY. (2009). The effects of lead exposure on placental NF-kappaB expression
and the consequences for gestation. Reprod Toxicol 27: 190-195.

http://dx.doi.org/.1.0..1.0.1.6/i.reprotox.2008..1.2.006.

Wells. EM: Navas-Acien. A: Herbstman. IB: Apelberg. BJ: Silbergeld. EK: Caldwell. KL: Jones. RL: Halden. RU:
Witter. FR: Goldman. LR. (2011). Low level lead exposure and elevations in blood pressure during
pregnancy. Environ Health Perspect 119: 664-669. http://dx.doi.org/10.1289/ehp. .1.002666.

Weston. HI: Weston. DP: Allen. JL: Corv-Slecht (2014). Sex-dependent impacts of low-level lead exposure
and prenatal stress on impulsive choice behavior and associated biochemical and neurochemical
manifestations. Neurotoxicology 44: 169-183. http://dx.doi.org/.1.0. .1.0.1.6/i. nenro.20.1.4.06.0.1.3.

Wide. M: Nilsson.	79). Interference of lead with implantation in the mouse: A study of the surface

ultrastructure of blastocysts and endometrium. Teratology 20: 101-113.

http://dx.doi.org/10.1002/tera. .1.420200.1. .1.4.

Williams. PL: Betlavia. A: Korrick. SA: Burns. JS: Lee. MM: Sergevev. O: Hauser. R. (2019). Blood lead levels and
timing of male sexual maturity: A longitudinal study of Russian boys. Environ Int 125: 470-477.

http://dx.doi.org/10. .1.016/i.envint.201.9.0.1. .070.

Williams. PL: Minguez-Alarcon. L: Korrick.	¦. MM: Plaku-Alakbarova. B: Burns. JS: Smigulina. L: Dikov.

Y: Abou Ghavda. R: Hauser. R: Sergevev. O. (2022). Association of peripubertal blood lead levels with
reproductive hormones and semen parameters in a longitudinal cohort of Russian men. Hum Reprod 37:
848-858. http://dx.doi.org/10.1093Ainm.rep/deab288.

8-258


-------
Williams. PL; Sergevev. O; Lee. MM; Korrick. SA; Burns. JS; Humblet. O; DelPrato. J; Revich. 6; Hauser. R.

(2010). Blood lead levels and delayed onset of puberty in a longitudinal study of Russian boys. Pediatrics
125: el088-el096. http://dx.doi.org/10.1542/peds.2009-2575.

Woods. MM; Lanphear. BP; Braun. JM; MeCandtess. LC. (2017). Gestational exposure to endocrine disrupting
chemicals in relation to infant birth weight: A Bayesian analysis of the HOME Study. Environ Health 16:
115. http://dx.doi.org/10.1186/sl2940-017-0332-3.

Wu. SZ; Xu. HY; Chen. Y; Che	ti. QL; Tan. M.H; Zhang. MM. (2021). Association of blood lead levels with

preeclampsia: A cohort study in China. Environ Res 195: 110822.

http://dx.doi.org/10. .1.016/i.envres.202.1.. .1. .1.0822.

Wu. T; Buck. GM; Mendota. P. (2003). Blood lead levels and sexual maturation in U.S. girls: The Third National
Health and Nutrition Examination Survey, 1988-1994. Environ Health Perspect 111: 737-741.

http://dx.doi.org/10.1289/elip.6008.

Xie. J; Yu. J; Fan. YS; Zhao. X; Su. JM; Meng. Y; Wu. Y; Uddin. MB; Wang. CH; Wang. ZS. (2020). Low dose
lead exposure at the onset of puberty disrupts spermatogenesis-related gene expression and causes
abnormal spermatogenesis in mouse. Toxicol Appl Pharmacol 393: 114942.
http://dx.doi.Org/10.lQ16/i.taap.202Q.114942.

Xie. X; Ding. GD; Cui. C; Chen. LM; Gao. Y; Zhou. YJ; Shi. R; Tian. Y. (2013). The effects of low-level prenatal
lead exposure on birth outcomes. Environ Pollut 175: 30-34.

http://dx.doi.org/10. .1.016/i.envpoL201.2.1.2.0.1.3.

Xu. R; Meng. X; Pang. Y; An. H; Wang. B; Zhang. L; Ye. R; Ren. A; Li. Z; Gong. J. (2022a). Associations of
maternal exposure to 41 metals/metalloids during early pregnancy with the risk of spontaneous preterm
birth: Does oxidative stress or DNA methylation play a crucial role? Environ Int 158: 106966.
http://dx.doi.Org/10.1016/i.envint.2021.106966.

Xu. S; Hansen. S; Sripada. K; Aarstand. T; Horvat. M; Mazei. D; Alvarez. MV; Odtand. J0. (2022b). Maternal
blood levels of toxic and essential elements and birth outcomes in Argentina: The EMASAR study. Int J
Environ Res Public Health 19: 3643. http://dx.doi.org/10.3390/iierphl.9063643.

Xu. XI; Yang. H; Chen. AM; Zhou. YL; Wu. KS; Liu. JX; Zhang. YL; Hno. X. (2012). Birth outcomes related to
informal e-waste recycling in Guiyu, China. Reprod Toxicol 33: 94-98.

http ://dx. do i .o rg/ .1.0. .1.0.1.6/i. repro tox.	.006.

Yang. X; Li. Y; Li. J; Bao. S; Zhou. A; Xu. S; Xia. W. (2020). Associations between exposure to metal mixtures and
birth weight. Environ Pollut 263: 114537. http://dx.doi.Org/10.101.6/i.envpol.2020..1.14537.

Yao. 0; Zhou. G; Xu. M; Dai. J; Qian. Z; Cai. Z; Zhang. L; Tan. Y; Hn. R. (2019). Blood metal levels and serum

testosterone concentrations in male and female children and adolescents: NHANES 2011-2012. PLoS ONE
14: e0224892. http://dx.doi.Org/.l.0.1371/ionrnaLpone.0224892.

Ye. S; Chung. HW; Jeong. K; Sung.	Park. SY; Kim. H; Ha. EH. (2017). Blood cadmium and volume of

uterine fibroids in premenopausal women. Ann Occup Environ Med 29: 22.
http://dx.doi.org/10.1186/s40557-017-0178-8.

Yu. YX; Gao. MM; Wang. XP; Giio. YH; Pang. YM; Yan. HN; Hao. YX; Zhang. YL; Zhang. L; Ye. RW; Wang. B;
Li. ZW. (2019). Recommended acceptable levels of maternal serum typical toxic metals from the
perspective of spontaneous preterm birth in Shanxi Province, China. Sci Total Environ 686: 599-605.
http://dx.doi.Org/10.1016/i.scitotenv.2019.05.413.

Zhang. Z; Yu. J; Xie. J; Lin. D; Fan. Y; Ma. H; Wang. C; Hong. Z. (2021). Improvement roles of zinc

supplementation in low dose lead induced testicular damage and glycolytic inhibition in mice. Toxicology
462: 152933. http://dx.doi.Org/10.1016/i.tox.2021.152933.

Zhao. ZH; Dn. KJ; Wang. T; Wang. JY; Cao. ZP; Chen. XM; Song. H; Zheng. G; Shen. XF. (2021). Maternal lead
exposure impairs offspring learning and memory via decreased GLUT4 membrane translocation. Front Cell
Dev Biol 9: 648261. http://dx.doi.org/10.3389/fcelL2021.648261.

8-259


-------
Zheng. Y; Lin. PIP; Williams. PL; Weisskopf. MG; Cardenas. A; Rifas-Shiman. SL; Wright. RO;

A ma rasi ri warde na. C; Henn. BC; Hivert. MF; Oken. E; James-Todd. T. (2021). Early pregnancy essential
and non-essential metal mixtures and gestational glucose concentrations in the 2nd trimester: Results from
Project Viva. Environ Int 155: 106690. http://dx.doi.org/10..1.016/i.envint.202.1...1.06690.

Zhou. CC; He. YO; Gao. ZY: Wn. MO; Yan. CH. (2020). Sex differences in the effects of lead exposure on growth
and development in young children. Chemosphere 250: 126294.
http://dx.doi.Org/10.1016/i.chemosphere.2020.126294.

Zhou. L; Liang. K; Li. M; Rong. C; Zheng. J; Li. .1. (2021a). Metal elements associate with in vitro fertilization
(IVF) outcomes in 195 couples. J Trace Elem Med Biol 68: 126810.

http://dx.doi.org/10. .1.016/j.jtemb.2Q2.1.. .1.268.1.0.

Zhou. Z; Chen. G; Li. P; Rao. J; Wang. L; Yu. D; Lin. D; Fan. D; Ye. S; Wu. S; Gou. X; Wang. H; Guo. X; Lin. L;
Suo. D; Liu. Z. (2021b). Prospective association of metal levels with gestational diabetes mellitus and
glucose: A retrospective cohort study from South China. Ecotoxicol Environ Saf 210: 111854.

http://dx.doi.org/10. .1.016/i.ecoenv.2020. .1. .1. .1.854.

8-260


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