Environmental Research 158 (2017) 583–589

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Environmental Research
journal homepage: www.elsevier.com/locate/envres

Micronutrients during pregnancy and child psychomotor development: MARK

Opposite effects of Zinc and Selenium
⁎
Kinga Polanskaa, , Wojciech Hankea, Anna Krola, Jolanta Gromadzinskab, Renata Kurasb,
Beata Janasikb, Wojciech Wasowiczb, Fiorino Mirabellac, Flavia Chiarottic, Gemma Calamandreic
a
Department of Environmental Epidemiology, Nofer Institute of Occupational Medicine, Lodz, Poland
b
Department of Biological and Environmental Monitoring, Nofer Institute of Occupational Medicine, Lodz, Poland
c
Center for Behavioral Sciences and Mental Health, National Institute of Health, Rome, Italy

A R T I C L E I N F O A B S T R A C T

Keywords: Studies on the impact of micronutrient levels during different pregnancy periods on child psychomotor functions
Zinc are limited. The aim of this study was to evaluate the association between maternal plasma concentrations of
Selenium selected micronutrients, such as: copper (Cu), zinc (Zn), selenium (Se), and child neuropsychological develop-
Copper ment. The study population consisted of 539 mother-child pairs from Polish Mother and Child Cohort
Prenatal period
(REPRO_PL). The micronutrient levels were measured in each trimester of pregnancy, at delivery and in the cord
Child neuropsychological development
blood. Psychomotor development was assessed in children at the age of 1 and 2 years using the Bayley Scales of
Infant and Toddler Development. The mean plasma Zn, Cu and Se concentrations in the 1st trimester of preg-
nancy were 0.91 ± 0.27 mg/l, 1.98 ± 0.57 mg/l and 48.35 ± 10.54 μg/l, respectively. There were no statisti-
cally significant associations between Cu levels and any of the analyzed domains of child development. A po-
sitive association was observed between Se level in the 1st trimester of pregnancy and child language and motor
skills (β=0.18, p=0.03 and β=0.25, p=0.005, respectively) at one year of age. Motor score among one-year-
old children decreased along with increasing Zn levels in the 1st trimester of pregnancy and in the cord blood
(β=−12.07, p=0.003 and β=−6.51, p=0.03, respectively). A similar pattern was observed for the association
between Zn level in the 1st trimester of pregnancy and language abilities at one year of age (β=−7.37,
p=0.05). Prenatal Zn and Se status was associated with lower and higher child psychomotor abilities, respec-
tively, within the first year of life. Further epidemiological and preclinical studies are necessary to confirm the
associations between micronutrient levels and child development as well as to elucidate the underlying me-
chanisms of their effects.

1. Introduction pathways and structural components (Georgieff, 2007; Georgieff and

Pregnancy is a period of increased metabolic demands mainly due to
the changes in a woman's physiology and requirements of a growing
fetus. Nutrients and growth factors regulate brain development during
fetal and early postnatal life. The developing brain is particularly vul-
nerable to nutritional insults as the rapid trajectory of several neuro-
logic processes, including synapse formulation and myelination
(Georgieff, 2007; Georgieff and Rao, 2001; Dobbing, 1990; Rao and
Georgieff, 2000; Kretchmer et al., 1996; Thompson and Nelson, 2001).
Thus, certain nutrients, such as: proteins, essential fatty acids, iron,
zinc, copper, iodine, selenium, vitamin A, choline and folates, are
especially significant for the developing brain (Nyaradi et al., 2013).
Effects of any nutrient deficiency or its excess on brain development
is a function of its requirement for a nutrient in specific metabolic
Rao, 2001; Dobbing, 1990; Rao and Georgieff, 2000; Kretchmer et al.,
1996; Thompson and Nelson, 2001). Zinc (Zn) is an essential element
with a multitude of biological functions, which plays an important role
in brain development and synaptic plasticity (Shah and Sachdev, 2006;
Frederickson and Danscher, 1990). In particular, Zn is not only a
component of several enzymes but it is found in synaptic vesicles in
areas of the brain endowed with high plasticity such as cortex and
hippocampal mossy fibers. Zn is also involved in the metabolism of
thyroid hormones, hormone transportation, receptor binding as well as
in the metabolism of neurotransmitters (Shah and Sachdev, 2006;
Frederickson and Danscher, 1990; Nakashima and Dick, 2009; Morley
et al., 1980; Golub et al., 1995). Studies have shown an association
between prenatal Zn deficiency and infant psychomotor development
(Yang et al., 2013; Boroujeni et al., 2009; Cetin et al., 2010; Leung

⁎
Corresponding author.
E-mail address: kinga.polanska@imp.lodz.pl (K. Polanska).

http://dx.doi.org/10.1016/j.envres.2017.06.037
Received 14 January 2017; Received in revised form 4 June 2017; Accepted 29 June 2017
0013-9351/ © 2017 Elsevier Inc. All rights reserved.
K. Polanska et al. Environmental Research 158 (2017) 583–589

et al., 2011; Georgieff, 2007). On the other hand, there is growing Table 1
evidence suggesting that excess of Zn can exert neurotoxic action, lead Characteristics of the exposure and outcome variables.
to cellular damage in vitro (Yang et al., 2013; Zhu et al., 2012) as well
Variables Mean SD Min Max
as in vivo (Kong et al., 1998) and can be associated with decreased
child psychomotor development (Yang et al., 2013; Hamadani et al., Zn concentrations (mg/l)
2002). in the 1st trimester of pregnancy (N=392) 0.91 0.27 0.37 2.41
in the 2nd trimester of pregnancy (N=304) 0.78 0.27 0.23 1.96
Copper (Cu) is a crucial cofactor for copper-containing enzymes that
in the 3rd trimester of pregnancy (N=299) 0.74 0.21 0.32 1.45
function in a number of important processes, including energy pro- in maternal blood at delivery (N=284) 0.77 0.30 0.26 3.20
duction, oxidant defense, extracellular matrix (ECM) protein cross- in cord blood (N=290) 1.08 0.30 0.24 2.64
linking, immune function, blood cell maturation, neuropeptide and Cu concentrations (mg/l)
catecholamine synthesis, myocardial contractility, iron mobilization in the 1st trimester of pregnancy (N= 392) 1.98 0.57 0.55 4.88
in the 2nd trimester of pregnancy (N=307) 2.36 0.59 0.81 4.52
and trafficking (Uriu-Adams et al., 2010; Turski and Thiele, 2009;
in the 3rd trimester of pregnancy (N=299) 2.55 0.53 0.37 5.00
Gambling et al., 2011). The existing studies indicate that balance be- in maternal blood at delivery (N=288) 2.39 0.65 0.66 4.59
tween Cu and iron assures proper neurocognitive and neurobehavioral in cord blood (N=290) 0.59 0.28 0.14 1.72
development (Cetin et al., 2010; Gambling et al., 2011, 2008; Georgieff, Se concentrations (µg/l)
in the 1st trimester of pregnancy (N=410) 48.35 10.54 16.12 91.40
2007; Leung et al., 2011; Uriu-Adams et al., 2010; Beard et al., 2003;
in the 2nd trimester of pregnancy (N=151) 42.25 9.08 14.82 69.68
Penland and Prohaska, 2004; Gybina and Prohaska, 2006; Beard, 2008; in the 3rd trimester of pregnancy (N=130) 37.29 9.79 14.16 61.70
Prohaska and Gybina, 2005). in maternal blood at delivery (N=310) 38.44 11.79 13.83 86.29
The well-known biological role of selenium (Se) is mainly associated in cord blood (N=311) 31.07 8.22 13.81 56.31
with selenoproteins, which are involved in the antioxidant defence Composite score for the one-year-old children
N=501
system and thyroid hormones metabolism (Rayman, 2012; Jablonska
Cognitive 106.2 10.7 80.0 145.0
et al., 2013; Roman et al., 2014; Holmgren and Lu, 2010). Studies in- Language 107.8 13.0 68.0 141.0
dicate that Se can have impact on child neuropsychological develop- Motor 104.4 13.4 73.0 151.0
ment (Yang et al., 2013; Skröder et al., 2014). In our previous analysis Composite score for the two-year-old children
N=341
prenatal Se status was associated positively with a child's psychomotor
Cognitive 110.2 16.1 80.0 145.0
abilities within the first two years of life (Polanska et al., 2016b). Language 101.7 12.7 74.0 144.0
On average, for adults, Se intake of 70 μg/day (2014), Cu intake of Motor 111.5 14.5 73.0 154.0
1.3 mg/day (2015) and Zn intake of 7.5 mg/day (2014) is re-
commended by the European Food Safety Authority, with increasing Zn-zinc, Cu-copper, Se-selenium.
requirements among pregnant and lactating women (European Food
Safety Authority). The nationally representative dietary survey data weeks of single pregnancy, no assisted conception, no pregnancy
from eight European countries, including Poland, has highlighted that complications and no chronic diseases.
there is a risk of low intakes of essential trace elements in specific po- Based on the study protocol, the women were interviewed once in
pulations and age groups (Mensink et al., 2013). That analysis has in- each trimester of pregnancy to collect and update socio-demographic
dicated that 1.1% of women in Poland had Cu intake below the lower data, medical and reproductive history as well as information about
reference nutrient intake (LRNI - intake value below which it is unlikely environmental, lifestyle and occupational exposure. During each visit
that normal health can be maintained over longer periods) and 13.1% and after delivery, biological samples (including saliva, urine, blood,
below the estimated average requirement (EAR – the intake adequate hair and cord blood) were collected.
for 50% of the population). These estimates for Zn were: 4.6% and A child's exposure, health status and psychomotor development
16.6%, respectively. Mean Se contents in the daily food rations col- were evaluated at one year of age and the assessment was repeated
lected from various public canteens and a group of students ranged when the child reached the age of 24 months (Polanska et al., 2011).
from 20 to 59 µg/day (Jablonska et al., 2013). The current analysis was restricted to 539 children who have been
The existing studies evaluating influence of Zn, Cu and Se levels on examined for their neuropsychological development. Among them 303
children's development have produced conflicting results, mainly be- children had both assessments (at one and two years of age), 198 were
cause of the lack of a valid indicator and assessment of these micro- examined only at around the 12th month of age and 38 only at the 24th
nutrients. Therefore, the aim of this study was to evaluate the impact of month of age (Table 1). The following factors were responsible for
the micronutrients: Cu, Zn, Se, measured in the blood collected in losses in the follow-up: refusal, child health problems, unknown address
pregnancy and in the cord blood, on psychomotor development of or telephone number and unknown reasons (Polanska et al., 2016a).
children enrolled in the prospective Polish Mother and Child Cohort The study was approved by the Ethical Committee of the Nofer
study - REPRO_PL. Institute of Occupational Medicine, Lodz, Poland and a written consent
was obtained from all the study subjects.
2. Methods
2.2. Micronutrients assessment
2.1. Study design and population
Blood samples were collected from each woman during the first
The analyses were based on the mother-child pairs from REPRO_PL (8th−12th week of pregnancy), second (20th–24th week of preg-
cohort - a multicenter prospective cohort established in 2007 that ex- nancy), third (30th–34th week of pregnancy) trimester of pregnancy, at
amines the relationship between environmental (including heavy me- delivery and from the cord, using a venoject system free from trace
tals, phthalates, polycyclic aromatic hydrocarbons) as well as lifestyle/ elements with lithium heparin as an anticoagulant. After centrifugation,
psychosocial determinants (including smoking, alcohol consumption, the plasma was collected and frozen at −20 °C until the analysis.
stress, family functioning, BMI, physical activity, microelements and Plasma Zn and Cu concentrations were analyzed by means of the flame
vitamins) and multiple aspects of development and health of a child atomic absorption spectrometry (FAAS) (Agarwal and Henkon, 1985).
(Polanska et al., 2009, 2011, 2016a). The detailed description of the This method had been validated using the reference material (lyophi-
cohort methodology has been published previously (Polanska et al., lized human reference serum samples of Seronorm from Nycomed
2009, 2011, 2016a). Briefly, the women were invited to participate in Pharma AS, Oslo, Norway) and through participation in the inter-
the cohort if they fulfilled the following inclusion criteria: up to 12 laboratory comparison trials. Measurements of plasma Se concentration

584
K. Polanska et al.
were performed using the graphite furnace atomic absorption spectro-
metry (GFAAS) on a Unicam Solar 989 QZ apparatus with Zeeman ef-
fect background corrector, in accordance with the modified method of
Nève et al. (Neve and Molle, 1986; Neve et al., 1987). Accuracy of the
method for Se determination was verified using the internal quality
control of the certified reference material BCR-637 (IRMM, Belgium),
where reference value and measured concentration were: 81.0 µg/l (in
the range 74–88 µg/l) and 82.5 ± 0.7 µg/l, respectively, and external
quality control of the German External Quality Assessment Scheme (G-
EQUAS) for analyses in biological materials.
2.3. Child psychomotor assessment
The Bayley Scales of Infant and Toddler Development (Bayley 3rd
edition) was used to assess children's neuropsychological development
(including cognitive, language and motor functions) at one and two
years of age. Details regarding child psychomotor assessment have been
published elsewhere (Polanska et al., 2011, 2016b).
2.4. Covariates
The following covariates were evaluated: parental age and educa-
tion; marital status; socio-economic status (SES); child gender; major
pregnancy complications which appeared after inclusion into the study;
type of delivery; gestational age and birth outcomes; breastfeeding;
number of siblings; day care attendance; maternal cigarette smoking
and alcohol consumption during pregnancy as well as child environ-
mental tobacco smoke (ETS) exposure within the first two years of live.
Details regarding socio-demographic and birth outcome data collection
as well as prenatal/postnatal ETS exposure have been described else-
where (Polanska et al., 2016b; Stragierowicz et al., 2013). In the cur-
rent analysis, the women whose cotinine level in saliva was equal to or
higher than 10 ng/ml were defined as active smokers (SRNT
Subcommittee on Biochemical Verification, 2002).
2.5. Statistical analysis
Levels of microelements in each trimester of pregnancy, at delivery
and in cord blood and cognitive, language and motor developmental
scores at 1 and 2 years of age are summarized by means, standard
deviation, and minimum and maximum values in the overall group of
children.
Correlation between microelement levels and psychomotor devel-
opmental scores (cognitive, language and motor) was examined by the
Pearson linear correlation coefficient. The multivariate linear regres-
sion was performed to assess the effect of Zn, Cu and Se levels measured
during the 1st trimester of pregnancy and in cord blood, on child
cognitive, language and motor scores at 1 and 2 years of age, and on the
developmental score variation between 1 and 2 years of age. Following
the study protocol, data on micronutrients was theoretically available
for each visit; however, as a result of organizational (i.e., the lack of
samples or usage of the samples for other purposes) and financial rea-
sons (in that case randomly selected samples were evaluated), not all
the samples were analyzed for micronutrient levels. In particular, the
number of samples collected during the 2nd and the 3rd trimesters of
pregnancy for which micronutrients were available was much lower,
thus, micronutrient levels measured at these time points were not in-
cluded in the multivariate analyses. The multivariate model included
confounders significant at 0.1 level. As for education, a mother and a
father's data were highly correlated (r > 0.5; p < 0.05), so only ma-
ternal age and education were selected and included in the model. The
final multivariate models were performed on 239 subjects at 12 months
assessment and 168 subjects at 24 months assessment, for which there
were both Zn, Cu and Se in the 1st trimester of pregnancy and in the
cord blood, and the confounding variables that were known to affect
child's psychomotor development: examiner, mother age, mother
585
Environmental Research 158 (2017) 583–589
education, child gender and maternal smoking status during pregnancy
based on the cotinine level. The analysis focusing on the effect of mi-
cronutrient levels during pregnancy on the change in cognitive, lan-
guage and motor scores between 1 and 2 years of age applied to all the
children undergoing both evaluations. An examiner assessing devel-
opment at one year of age and an examiner performing assessment at
two years of age were included as covariates in the model. In addition,
the study population was divided in quartiles according to the micro-
nutrient levels. Low (less than 1st quartile), medium (between 1st and
3rd quartile) and high (more than 3rd quartile) groups were compared
in order to evaluate the effect of micronutrients on a child's develop-
ment (result of the analysis is presented in the Supplementary mate-
rials).
The variance inflation factor (VIF) was computed for any variable in
each model to verify the presence of multicollinearity among ex-
planatory variables. Regression coefficients are reported together with
their standard errors. STATA Statistical Software (Release 8.1) was used
for the statistical analyses.
3. Results
3.1. Parental and child characteristics
Parental and child characteristics are summarized in the
Supplementary materials (Tables S1, S2). A high percentage of the
women were married (75.0%), employed (84.8%) and had a university
degree (62.8%). Alcohol consumption during pregnancy was indicated
by 8.5% of the women and, based on the cotinine level in saliva, 15.1%
were classified as smokers. About 7% of the children attended day care
at one and 23% at two years of age. ETS exposure after birth was noted
for 35.8% of the children.
The mean composite scores for cognitive, language and motor de-
velopment were on an average or high average level (Table 1). A po-
sitive correlation was observed for each subscale of the Bayley test
results for one year assessment (p≤0.001) and between cognitive and
language as well as motor and language functions (p≤0.001) for two-
year assessment (Table 3). The correlation was weaker for comparisons
performed between one and two years of age.
3.2. Micronutrient concentrations during pregnancy
The micronutrient concentrations in the blood collected in each trime-
ster of pregnancy, at delivery and in the cord blood are presented in Table 1.
Mean Zn concentration was higher in cord blood (1.08 ± 0.3 mg/l) and in
the 1st trimester of pregnancy (0.91 ± 0.27 mg/l) than in the other mea-
surement periods (p < 0.001). Mean plasma Cu concentration was slightly
lower in the 1st trimester of pregnancy (1.98 ± 0.57 mg/l) and remained
fairly constant during the two other periods of pregnancy and in maternal
blood collected at delivery (2.36 ± 0.59 mg/l in the 2nd trimester,
2.55 ± 0.53 mg/l in the 3rd trimester and 2.39 ± 0.65 mg/l at delivery).
Cu concentration was significantly lower in the cord blood than in preg-
nancy period (0.59 ± 0.28 mg/l) (p < 0.001). Mean Se concentrations de-
creased throughout pregnancy and at delivery (from 48.35 ± 10.54 μg/l in
the 1st trimester to 38.44 ± 11.79 μg/l at delivery) (p < 0.001). Correla-
tions between different microelements as well as their concentrations in
different time periods are presented in Table 2.
3.3. Association between micronutrient concentrations and child
psychomotor development
Table 4 presents the association between maternal micronutrient
levels and child psychomotor development with adjustment for con-
founders. There were no statistically significant associations between
Cu levels in the blood collected during the 1st trimester of pregnancy
and in the cord blood and any of the analyzed domains of child de-
velopment. A positive association was observed between Se level in the
K. Polanska et al. Environmental Research 158 (2017) 583–589

Table 2
Pearson correlation (1st line=coefficient, 2nd line=sample size) between the micronutrient levels: zinc, copper, selenium in each trimester of pregnancy, at delivery and in the cord
blood.

Zn (A) Zn (B) Zn (C) Zn (D) Zn (E) Se (A) Se (B) Se (C) Se (D) Se (E) Cu (A) Cu (B) Cu (C) Cu (D) Cu (E)

Zn(A) 1
392
Zn(B) 0.31** 1
301 304
Zn(C) 0.46** 0.49** 1
296 288 299
Zn(D) 0.16* 0.25** 0.29** 1
283 253 251 284
Zn(E) 0.21** 0.21** 0.26** 0.29** 1
289 258 256 279 290
Se(A) 0.12* 0.04 0.12* 0.12** 0.08 1
390 301 296 283 289 410
Se(B) −0.12 0.21* 0.13 0.18* −0.02 0.35** 1
151 150 144 132 135 151 151
Se(C) −0.20* 0.17 −0.005 0.10 −0.04 0.38** 0.50** 1
130 125 129 119 120 130 125 130
Se(D) 0.23** 0.23** 0.35** 0.20** 0.03 0.32** 0.27* 0.47** 1
306 254 252 278 279 310 135 121 310
Se(E) 0.14* 0.11 0.11 0.06 0.17* 0.14* 0.08 0.15 0.35** 1
308 253 251 272 280 311 133 118 302 311
Cu(A) 0.01 −0.005 0.002 0.04 −0.07 0.04 0.09 0.05 0.06 0.08 1
302 301 296 283 390 390 151 130 306 308 392
Cu(B) 0.07 0.10 0.23** 0.19* 0.12* 0.12* 0.10 0.13 0.15* −0.04 0.34** 1
304 303 291 255 261 304 150 126 256 256 304 307
Cu(C) 0.04 0.02 0.07 0.04 −0.001 0.02 −0.004 0.13 −0.04 0.07 0.17* 0.27** 1
296 288 299 251 256 296 144 129 252 251 296 291 299
Cu(D) 0.003 −0.02 −0.10 0.05 0.01 0.008 0.01 0.03 −0.05 0.03 0.18* 0.19* 0.44* 1
287 256 255 282 284 287 133 119 277 277 287 259 255 288
Cu(E) 0.20** 0.27** 0.30** 0.26** 0.24** 0.17* −0.03 −0.04 0.14* 0.11 −0.05 0.07 0.14* 0.15* 1
289 258 256 279 290 289 135 120 279 280 289 261 256 284 290

A: blood collected between the 8th–12th weeks of pregnancy, B: blood collected between the 20th–24th weeks of pregnancy, C: blood collected between the 30th–34th weeks of
pregnancy, D: blood collected from mothers at delivery, E: cord blood; Zn-zinc, Cu-copper, Se-selenium.
Correlation coefficients significantly different from 0 are reported in bold: *, **=p < 0.05 or p < 0.01, respectively.

1st trimester of pregnancy and child language and motor skills
(β=0.18, p=0.03 and β=0.25; p=0.005, respectively) at one year of
age. Interestingly, motor scores among one-year-old children decreased
along with increasing Zn levels in the 1st trimester of pregnancy and in
the cord blood (β=−12.07, p=0.003 and β=−6.51, p=0.03, re-
spectively). A similar pattern was observed for the association between
Zn level in the 1st trimester of pregnancy and language abilities at one
year of age (β=−7.37, p=0.05). The analysis with the study popula-
tion divided in quartiles according to micronutrient levels indicated
similar results to these with micronutrients as continuous variables
(opposite impact of Zn and Se on child motor abilities at the age of 1
year) (Table S4). The analysis focusing on the impact of micronutrient
levels on child psychomotor development at the age of 2 years did not
give any significant results (with and without child developmental
scores at 1 year of age as additional covariates) (Tables S3 and 4).
An additional analysis regarding micronutrient levels during
pregnancy and the change in psychomotor development scores between
1 and 2 years of age is presented in Table 5. None of the results were
statistically significant (> 0.05).
4. Discussion
Our prospective cohort study indicated a positive association be-
tween Se level in the 1st trimester of pregnancy and child language and
motor skills at one year of age. In addition, motor and language scores
decreased along with increasing Zn levels and there were no statisti-
cally significant associations between Cu levels and any aspects of child
development.
Micronutrient concentrations are different in pregnant than in non-
pregnant women. Plasma Zn concentration begins to decline in early
pregnancy and continues to decline until delivery, when it is about 35%
below its level in non-pregnant women (Wasowicz et al., 1993; King,

Table 3
Pearson correlation between the Bayley test results for one- and two-year assessments.

Psychomotor abilities Assessment at the age of 1 Assessment at the age of 2

Cognitive Language Motor Cognitive Language Motor

Assessments at the age of 1 Cognitive 1

Language 0,25** 1
Motor 0,31** 0,43** 1
Assessments at the age of 2 Cognitive 0,13* −0,03 0,12* 1
Language 0,04 0,15* 0,08 0,49** 1
Motor −0,04 0,22** 0,16* 0,06 0,35** 1

For correlations within the assessment at one year N=501; for correlations between the assessment at one and two years N=303; for correlations within the assessment at two years
N=341.
* p < 0.05.
** p≤0.001.

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K. Polanska et al. Environmental Research 158 (2017) 583–589

Table 4
Micronutrient levels during pregnancy and child psychomotor development–the multivariate model.

Microelements 1-year old children β (95% CI) (p) 2-year old children β (95% CI) (p)

Cognitive Language Motor Cognitive Language Motor

Zn (1st trimester) (mg/l) −1.74 −7.37 −12.07 1.36 3.71 −0.60

(−8.33;4.84) (−14.73;−0.01) (−20.12;−4.02) (−9.46;12.19) (−6.14;13.56) (−10.61;9.42)
(0.60) (0.05) (0.003) (0.80) (0.46) (0.91)
Zn (cord blood) (mg/l) 0.11 −3.77 −6.51 1.70 −0.19 −5.68
(−4.79;5.01) (−9.25;1.71) (−12.50;−0.52) (−6.72;10.12) (−7.85;7.48) (−13.47;2.11)
(0.97) (0.18) (0.03) (0.69) (0.96) (0.15)
Cu (1st trimester) (mg/l) −0.14 1.38 0.32 −1.15 2.40 −1.47
(−2.45;2.17) (−1.21;3.96) (−2.5–3.15) (−4.75;2.45) (−0.88;5.68) (−4.80;1.86)
(0.91) (0.30) (0.82) (0.53) (0.15) (0.39)
Cu (cord blood) (mg/l) −1.03 1.90 5.06 1.70 0.69 0.78
(−6.03;98) (−3.70;7.49) (−1.05;11.18) (−6.22;9.63) (−6.52;7.90) (−6.55;8.11)
(0.69) (0.50) (0.10) (0.67) (0.85) (0.83)
Se (1st trimester) (µg/l) 0.11 0.18 0.25 0.22 0.07 0.15
(−0.03;0.25) (0.02;0.34) (0.08;0.42) (−0.02;0.47) (−0.15;0.29) (−0.07;0.38)
(0.13) (0.03) (0.005) (0.07) (0.52) (0.18)
Se (cord blood) (µg/l) 0.005 0.10 0.17 −0.06 0.09 0.07
(−0.16;0.17) (−0.08;0.29) (−0.03;0.37) (−0.32;0.19) (−0.14;0.32) (−0.16;0.31)
(0.95) (0.28) (0.10) (0.62) (0.44) (0.55)

Adjusted for: examiner, maternal age, maternal education, child gender and maternal smoking status during pregnancy based on the cotinine level.
For the assessment at the age of 1year N=239.
For the assessment at the age of 2 years N=168.
Data is reported as regression coefficients (β), 95% Confidence Intervals (95% CI), p values (p).

Table 5 earlier study performed in Poland, cord plasma Zn level was 0.8 mg/l
Micronutrient levels during pregnancy and the change in psychomotor developmental (Wasowicz et al., 1993). The other study in Poland (although not cov-
scores between 1 and 2 years of age–the multivariate model.
ering pregnant women) has indicated the mean intake (analytical data)
Microelements Change of a child's developmental score between 1 and 2 of Zn of 8.9 mg/day (5.3 mg/day calculated from 24-h diet recalls),
years of age β (95% CI) (p) which is comparable to the mean Zn intake in Spain (7.6 mg/day) and
lower than that in Germany (10.9 mg/day) (Jablonska et al., 2013;
Cognitive Language Motor Flynn et al., 2009). Similar results have been also obtained for 18−60
Zn (1st trimester) −2.00 6.77 0.70
year-old females in a nationally representative dietary survey data from
(mg/l) (9.51;13.51) (−3.82;17.36) (−9.50;10.91) eight European countries (Mensink et al., 2013).
(0.73) (0.21) (0.89) Based on the existing data, intake of Se varies hugely worldwide
Zn (cord blood) 0.85 1.68 −1.95 from low to even toxic concentrations with the mean values of 40 μg/
(mg/l) (−8.60;10.29) (−6.84;10.20) (−10.17;6.28)
day in Europe, and 93 μg/day in the United States (Rayman, 2012). The
(0.86) (0.70) (0.64)
Cu (1st trimester) −1.99 3.26 −0.57 detailed discussion regarding Se levels observed in our cohort has been
(mg/l) (−5.90;1.91) (−0.26;6.77) (−3.94;2.80) published previously (Polanska et al., 2016b). It is worth noting that in
(0.32) (0.07) (0.74) Poland plasma Se level was 48 μg/l in the studies performed between
Cu (cord blood) 2.65 −0.93 −1.53 1981 and 1983 and above 30 μg/l in that performed between 1997 and
(mg/l) (−6.00;11.31) (−8.79;6.93) (−9.18;6.12)
(0.55) (0.82) (0.69)
1999 and between 2007 and 2009 (REPRO_PL study) (Zachara et al.,
Se (1st trimester) 0.11 0.07 0.12 1986; Jablonska et al., 2013; Wasowicz et al., 2003; Polanska et al.,
(µg/l) (−0.16;0.39) (−0.18;0.32) (−0.12;0.35) 2016b).
(0.42) (0.58) (0.33) In our study, mean plasma Cu concentration increased throughout
Se (cord blood) −0.05 0.07 −0.01
the pregnancy period (from 1.98 mg/l in the 1st trimester of pregnancy
(µg/l) (−0.33;0.22) (−0.17;0.32) (−0.25;0.23)
(0.70) (0.56) (0.94) to 2.36 ml/l in the 2nd and 2.55 mg/l in the 3rd trimester of preg-
nancy). This is consistent with the results reported in other studies on
Adjusted for: examiner at first and at second assessment, child gender, maternal age, pregnant women (Pathak, 2004). Studies have found that an increase in
maternal education, maternal smoking status during pregnancy based on the cotinine the Cu level during pregnancy is mainly in a bound form due to an
level and child developmental score (cognitive, language or motor, respectively) at 1 year
increase in the carrier proteins, ceruloplasmin in response to a stimu-
of age (N=149).
lation by maternal estrogens (Pathak et al., 2004; Martín-Lagos et al.,
Data is reported as regression coefficients (β), 95% Confidence Intervals (95% CI), p
values (p). 1998; Kalra et al., 1989; Dokumov, 1968). With regard to the mean Cu
intake in Poland, based on the female population, it was 1.1 mg/day,
2000; Cetin et al., 2010; Yang et al., 2013). Significant decrease of Zn which was similar to that observed in France (1.2 mg/day), The Neth-
concentration was also observed in our study (from 0.91 mg/l in the 1st erlands (1.1 mg/day), the UK (1.1 mg/day) and lower than that re-
trimester of pregnancy to 0.77 mg/l at delivery). This decline in Zn ported in Germany (2.2 mg/day) (Mensink et al., 2013; Flynn et al.,
levels has been attributed to hemodilution and/or different Zn affinity 2009). Jablonska et al. have indicated the mean intake of Cu to be
to plasma proteins as a consequence of hormonal changes during 1.4 mg/l (based on the analytical data) and 0.8 mg/day (based on the
pregnancy and it can result from active transportation of Zn from the data calculated from 24-h diet recalls) (Jablonska et al., 2013).
mother to the fetus (Wasowicz et al., 1993; King, 2000). The median Our findings support the existing evidence indicating a significant
level of cord plasma Zn (1.08 mg/l) in our study was similar to that role of Se in the brain and behavior development. We observed a po-
reported in Arctic Canada (1.1 mg/l) and higher than that reported in sitive association between Se level in the 1st trimester of pregnancy and
China (0.73 mg/l) (Butler Walker et al., 2006; Yang et al., 2013). In an child language and motor skills at one year of age. It is important that
the data concerning the effect of Se levels (also in cord blood) on child

587
K. Polanska et al.
psychomotor development at the age of two years are in the same di-
rection, although not statistically significant. In the study by Skröder
et al., similarly to our observations, an increase in the maternal Se
concentration has been associated with improvement in children's
language and psychomotor development (Skröder et al., 2014). What is
interesting is that in the study by Yang et al. low (< 100 µg/l) and high
(≥100 µg/l) levels of cord serum Se had negative effects on neonatal
neuropsychological development (Yang et al., 2013). However, in our
assessment, the maximum Se level in cord plasma was 56 μg/l, which is
much lower than the Se level observed by Yang et al. More details on
the impact of Se on child psychomotor development can be found in our
previous publication (Polanska et al., 2016b).
The existing studies evaluating the impact of Zn supplementation
during pregnancy as well as Zn level in the maternal and/or cord blood
on child development are not consistent. As an example, two studies
have found no effect of Zn supplementation during pregnancy on child
psychomotor development (Tamura et al., 2003; Caulfield et al., 2010),
while in two other studies, a negative effect of Zn supplementation on
developmental outcomes has been observed (Hamadani et al., 2002;
Wehby and Murray, 2008). In the study by Yang and coworkers the
cord serum Zn level ≥ 0.794 mg/l had an adverse effect on neonatal
neurobehavioral development (Yang et al., 2013). This is in agreement
with what we observed in our study – decreasing motor and language
scores along with increasing Zn levels. It is important to be aware of the
fact that in the present study, in about 88% of the cord blood samples
Zn level was higher than the threshold determined by Yang et al. (Yang
et al., 2013). To explain the negative effects of Zn on psychomotor
scores at one year, it is worth mentioning that Zn may have both
neuroprotective and neurotoxic properties. Specific mechanisms un-
derlying Zn neurotoxicity have been described by linking excess Zn to
oxidative stress and an increased rate of cell death (Morris and
Levenson, 2012; Yang et al., 2013). Furthermore, given the pivotal role
of Zn in glutamate neurotransmission, it can be expected that even
relatively slight changes in vesicular Zn levels during early neurode-
velopment may influence cortical/hippocampal synaptogenesis and
possibly later behavioural functions (Nakashima and Dyck, 2009).
We have not found statistically significant associations between Cu
levels and any aspects of child psychomotor development; however,
some studies in this field indicate that such associations exist, and that
balance between Cu and iron assures proper child neurocognitive and
neurobehavioral development (Cetin et al., 2010; Gambling et al.,
2011, 2008; Georgieff, 2007; Leung et al., 2011; Uriu-Adams et al.,
2010; Beard et al., 2003; Penland and Prohaska, 2004; Gybina and
Prohaska, 2006; Beard, 2008; Prohaska and Gybina, 2005).
An advantage of the current analysis is the assessment of micro-
nutrient status based on measurement of Zn, Cu and Se in the blood
over pregnancy period and not just on the questionnaire data regarding
diet and supplements. What is important is the fact that our analysis
evaluates the impact of all the three micronutrients (included in the
multivariate model) on child psychomotor development. Additionally,
in comparison to the existing studies, the current analysis takes ad-
vantage on inclusion of a variety of potential confounding factors and
the use of a well-standardized test evaluating neuropsychological ef-
fects in young children (Bayley 3rd edition).
Nevertheless, limitations of the study are also worth noting. The
main limitation concerns the lack of information on the dietary habits
of the women in the study in order to understand whether specific
dietary styles might be significantly associated with plasma levels of
specific micronutrients and with child psychomotor development. It has
to be pointed that all the nutrients are important for neuronal cell
growth and development (Georgieff, 2007). In our study we focused
only on Zn, Cu and Se and did not take into account general dietary
patterns of the pregnant women. We also did not include the intake of
protein, iron, iodine, folate, vitamins, choline and long-chain poly-
unsaturated fatty acids, which are also crucial for brain development. In
the current analysis correlation between micronutrients status and
588
Environmental Research 158 (2017) 583–589
maternal BMI was low, which indicates that they do not necessarily
reflect the overall nutritional status. It is worth mentioning that in our
cohort more than one-fifth of the women consumed multiple micro-
nutrient capsules (more than 20% of the population with Se and about
30% of the population with Zn as one of their components). However,
this neither influenced micronutrients status nor was associated with
any aspects of child psychomotor development (Polanska et al., 2016b).
The other limitation of the study is related to the fact that we did not
measure/or include internal exposure to neurotoxic elements such as
As, Cd, Pb, Hg. Additionally, although we measured micronutrient le-
vels in different pregnancy periods (for a substantial proportion of
children) the final sample, taking into account availability of the data,
was limited.
5. Conclusion
In conclusion, our study showed that prenatal Zn and Se statuses
were significantly associated with child psychomotor abilities within
the first years of life. It needs to be highlighted that micronutrient levels
in pregnant women depend on dietary sources and are influenced by
hormonal and metabolic changes throughout the pregnancy period. Our
results support the need of assuring a proper nutritional status of
women to prevent detrimental nutritional unbalance (Cetin et al.,
2010) and suggest that micronutrient supplementation during preg-
nancy should be considered with more caution, especially under op-
timal conditions. It would be of interest to explore the effects of preg-
nant women's diet on micronutrient levels and neuropsychological
development of their offspring. The effects which are reported here, and
those of Zn in particular, appear to be transient as they are no more
evident at 2 years of age. However, a follow-up of this population may
be useful to exclude effects on neuropsychological functions at later
ages. Altogether, further epidemiological and preclinical studies are
needed to confirm the association and elucidate the underlying me-
chanisms of these effects.
Acknowledgements
This study was partly funded by the National Science Centre under
the call JPI HDHL Nutrition and Cognitive Function (2015/17/Z/NZ7/
04273) and partly by the National Science Centre, Poland, under the
grant DEC-2014/15/B/NZ7/00998, by FP7 HEALS Grant N° 603946,
the Ministry of Science and Higher Education under grant agreement
no. 3068/7.PR/2014/2 and by CROME Grant N° LIFE12 ENV/GR/
001040.
Appendix A. Supplementary material
Supplementary data associated with this article can be found in the
online version at http://dx.doi.org/10.1016/j.envres.2017.06.037.
References
Agarwal, R.P., Henkon, R.L., 1985. A simple method for simultaneous estimation of zinc
and copper in erythrocytes. Biol. Trace. Elem. Res. 7 (4), 199–208. http://dx.doi.org/
10.1007/BF02989246.
Bayley 3rd edition. Retrieved from 〈http://www.pearsonclinical.com/childhood/
products/100000123/bayley-scales-of-infant-and-toddler-development-third-
edition-bayley-iii.html〉.
Beard, J., Erikson, K.M., Jones, B.C., 2003. Neonatal iron deficiency results in irreversible
changes in dopamine function in rats. J. Nutr. 133 (4), 1174–1179.
Beard, J.L., 2008. Why iron deficiency is important in infant development. J. Nutr. 138
(12), 2534–2536.
Boroujeni, S.T., Naghdi, N., Shahbazi, M., Farrokhi, A., Bagherzadeh, F., Kazemnejad, A.,
Javadian, M., 2009. The effect of severe zinc deficiency and zinc supplement on
spatial learning and memory. Biol. Trace Elem. Res. 130 (1), 48–61. http://dx.doi.
org/10.1007/s12011-008-8312-7.
Butler Walker, J., Houseman, J., Seddon, L., McMullen, E., Tofflemire, K., Mills, C.,
Corriveau, A., Weber, J.P., LeBlanc, A., Walker, M., Donaldson, S.G., Van Oostdam,
J., 2006. Maternal and umbilical cord blood levels of mercury, lead, cadmium, and
essential trace elements in Arctic Canada. Environ. Res. 100 (3), 295–318.
K. Polanska et al.
Caulfield, L.E., Putnick, D.L., Zavaleta, N., Lazarte, F., Albornoz, C., Chen, P., Dipietro,
J.A., Bornstein, M.H., 2010. Maternal gestational zinc supplementation does not in-
fluence multiple aspects of child development at 54 mo of age in Peru. Am. J. Clin.
Nutr. 92 (1), 130–136. http://dx.doi.org/10.3945/ajcn.2010.29407.
Cetin, I., Berti, C., Calabrese, S., 2010. Role of micronutrients in the periconceptional
period. Hum. Reprod. Update 16 (1), 80–95. http://dx.doi.org/10.1093/humupd/
dmp025.
European Food Safety Authority. Retrieved from (〈https://www.efsa.europa.eu/en/
topics/topic/drv〉).
Dobbing, J., 1990. Vulnerable periods in the developing brain. In: Dobbing, J. (Ed.),
Brain, behavior and iron in the infant diet. Springer, London, United Kingdom, pp.
1–25.
Dokumov, S.I., 1968. Serum copper and pregnancy. Am. J. Obstet. Gynecol. 101 (2),
217–222.
Flynn, A., Hirvonen, T., Mensink, G.B.M., Ocke, M.C., Serra-Majem, L., Stos, K., Szponar,
L., Tetens, I., Turrini, A., Fletcher, R., Wildemann, T., 2009. Intake of selected nu-
trients from foods, from fortification and from supplements in various European
countries. Food Nutr. Res. 1. http://dx.doi.org/10.3402/fnr.v53i0.2038.
Frederickson, C.J., Danscher, G., 1990. Zinc-containing neurons in hippocampus and
related CNS structures. Prog. Brain Res. 83, 71–84.
Gambling, L., Andersen, H.S., McArdle, H.J., 2008. Iron and copper, and their interactions
during development. Biochem. Soc. Trans. 36 (6), 1258–1261. http://dx.doi.org/10.
1042/BST0361258.
Gambling, L., Kennedy, C., McArdle, H.J., 2011. Iron and copper in fetal development.
Semin. Cell Dev. Biol. 22 (6), 637–644. http://dx.doi.org/10.1016/j.semcdb.2011.
08.011.
Georgieff, M.K., 2007. Nutrition and the developing brain: nutrient priorities and mea-
surement. Am. J. Clin. Nutr. 85 (2), 614–620.
Georgieff, M.K., Rao, R., 2001. The role of nutrition in cognitive development. In: Nelson,
C.A., Luciana, M. (Eds.), Handbook in developmental cognitive neuroscience. MIT
Press, Cambridge, MA, pp. 491–504.
Golub, M.S., Keen, C.L., Gershwin, M.E., Hendrickx, A.G., 1995. Developmental zinc
deficiency and behavior. J. Nutr. 125 (8), 2263–2271.
Gybina, A.A., Prohaska, J.R., 2006. Variable response of selected cuproproteins in rat
choroid plexus and cerebellum following perinatal copper deficiency. Genes Nutr. 1
(1), 51–59. http://dx.doi.org/10.1007/BF02829936.
Hamadani, J.D., Fuchs, G.J., Osendarp, S.J., Huda, S.N., Grantham-McGregor, S.M., 2002.
Zinc supplementation during pregnancy and effects on mental development and
behaviour of infants: a follow-up study. Lancet 360 (9329), 290–294. http://dx.doi.
org/10.1016/S0140-6736(02)09551-X.
Holmgren, A., Lu, J., 2010. Thioredoxin and thioredoxin reductase: current research with
special reference to human disease. Biochem. Biophys. Res. Commun. 396 (1),
120–124. http://dx.doi.org/10.1016/j.bbrc.2010.03.083.
Jablonska, E., Gromadzinska, J., Klos, A., Bertrandt, J., Skibniewska, K., Darago, A.,
Wasowicz, W., 2013. Selenium, zinc and copper in the Polish diet. J. Food Compos.
Anal. 31 (2), 259–265. http://dx.doi.org/10.1016/j.jfca.2013.05.016.
Kalra, R., Kalra, V.B., Sareen, P.M., Khandelwal, R., 1989. Serum copper and cer-
uloplasmin in pregnancy with anaemia. Indian J. Pathol. Microbiol. 32 (1), 28–32.
King, J.C., 2000. Determinants of maternal zinc status during pregnancy. Am. J. Clin.
Nutr. 71 (5), 1334–1343.
Kong, X., Liu, L., Sheng, X., 1998. Effects of excessive zinc in fodder on brain development
and abilities of learning and memory and their mechanisms in young rats. Zhonghua
Yu Fang. Yi Xue Za Zhi. 32 (4), 225–228.
Kretchmer, N., Beard, J.L., Carlson, S., 1996. The role of nutrition in the development of
normal cognition. Am. J. Clin. Nutr. 63 (6), 997–1001.
Leung, B.M.Y., Wiens, K.P., Kaplan, B.J., 2011. Does prenatal micronutrient supple-
mentation improve children's mental development? A systematic review. BMC
Pregnancy Childbirth 11, 1–12. http://dx.doi.org/10.1186/1471-2393-11-12.
Martín-Lagos, F., Navarro-Alarcón, M., Terrés-Martos, C., López-García de la Serrana, H.,
Pérez-Valero, V., López-Martínez, M.C., 1998. Zinc and copper concentrations in
serum from Spanish women during pregnancy. Biol. Trace Elem. Res. 61 (1), 61–70.
Mensink, G.B., Fletcher, R., Gurinovic, M., Huybrechts, I., Lafay, L., Serra-Majem, L.,
Szponar, L., Tetens, I., Verkaik-Kloosterman, J., Baka, A., Stephen, A.M., 2013.
Mapping low intake of micronutrients across Europe. Br. J. Nutr. 110 (4), 755–773.
http://dx.doi.org/10.1017/S000711451200565X.
Morley, J.E., Gordon, J., Hershman, J.M., 1980. Zinc deficiency, chronic starvation, and
hypothalamic-pituitary-thyroid function. Am. J. Clin. Nutr. 33 (8), 1767–1770.
Morris, D.R., Levenson, C.W., 2012. Ion channels and zinc: mechanisms of neurotoxicity
and neurodegeneration. J. Toxicol. 2012, 785647. http://dx.doi.org/10.1155/2012/
785647.
Nakashima, A.S., Dyck, R.H., 2009. Zinc and cortical plasticity. Brain Res. Rev. 59 (2),
347–373. http://dx.doi.org/10.1016/j.brainresrev.2008.10.003.
Neve, J., Chamart, S., Molle, L., 1987. Optimization of direct procedure for the de-
termination of selenium in plasma and erythrocytes using Zeeman effect atomic ab-
sorption spectroscopy. Trace Elem. Anal. Chem. Med. Biol. 4, 349–358.
Neve, J., Molle, L., 1986. Direct determination of selenium in human serum by graphite
furnace atomic absorption spectroscopy. Improvements due to oxygen ashing in
589
Environmental Research 158 (2017) 583–589
graphite tube and Zeeman effect background correction. Acta Pharmacol. Toxicol. 59
(7), 606–609.
Nyaradi, A., Li, J., Hickling, S., Foster, J., Oddy, W.H., 2013. The role of nutrition in
children's neurocognitive development, from pregnancy through childhood. Front
Hum. Neurosci. 7, 1–16. http://dx.doi.org/10.3389/fnhum.2013.00097.
Pathak, P., Kapil, U., Kapoor, S.K., Saxena, R., Kumar, A., Gupta, N., Dwivedi, S.N., Singh,
R., Singh, P., 2004. Prevalence of multiple micronutrient deficiencies amongst
pregnant women in a rural area of Haryana. Indian J. Pediatr. 71 (11), 1007–1014.
Penland, J.G., Prohaska, J.R., 2004. Abnormal motor function persists following recovery
from perinatal copper deficiency in rats. J. Nutr. 134 (8), 1984–1988.
Polanska, K., Hanke, W., Gromadzinska, J., Ligocka, D., Gulczynska, E., Sobala, W.,
Wasowicz, W., 2009. Polish mother and child cohort study- defining the problem, the
aim of the study and methodological assumption. Int. J. Occup. Med. Environ. Health
22 (4), 383–391. http://dx.doi.org/10.2478/v10001-009-0037-0.
Polanska, K., Hanke, W., Jurewicz, J., Sobala, W., Madsen, C., Nafstad, P., Magnus, P.,
2011. Polish mother and child cohort study (REPRO_PL)- methodology of follow-up
of the children. Int. J. Occup. Med. Environ. Health 24 (4), 391–398. http://dx.doi.
org/10.2478/s13382-011-0026-y.
Polanska, K., Hanke, W., Krol, A., Hanke, A., Waszkowska, M., Jacukowicz, A.,
Gromadzinska, J., Wasowicz, W., Jerzynska, J., Stelmach, W., Stelmach, I., 2016a.
Polish Mother and Child Cohort Study (REPRO_PL) - Methodology of the follow-up of
the children at the age of 7. Int. J. Occup. Med. Environ. Health 29 (6), 883–893.
http://dx.doi.org/10.13075/ijomeh.1896.00811.
Polanska, K., Krol, A., Sobala, W., Gromadzinska, J., Brodzka, R., Calamandrei, G.,
Chiarotti, F., Wasowicz, W., Hanke, W., 2016b. Selenium status during pregnancy
and child psychomotor development-Polish Mother and Child Cohort study. Pediatr.
Res. 79 (6), 863–869. http://dx.doi.org/10.1038/pr.2016.32.
Polish Mother and Child Cohort study RERPO_PL. Retrieved from (〈www.repropl.com〉).
Prohaska, J.R., Gybina, A.A., 2005. Rat brain iron concentration is lower following
perinatal copper deficiency. J. Neurochem. 93 (3), 698–705.
Rao, R., Georgieff, M.K., 2000. Early nutrition and brain development. In: Nelson, C.A.
(Ed.), The effects of early adversity on neurobehavioral development. Minnesota
Symposium on Child Psychology 31. Erlbaum Associates, Hillsdale, NJ, pp. 1–30.
Rayman, M.P., 2012. Selenium and human health. Lancet 379 (9822), 1256–1268.
http://dx.doi.org/10.1016/S0140-6736(11)61452-9.
Roman, M., Jitaru, P., Barbante, C., 2014. Selenium biochemistry and its role for human
health. Metallomics 6 (1), 25–54. http://dx.doi.org/10.1039/c3mt00185g.
Shah, D., Sachdev, H.P., 2006. Zinc deficiency in pregnancy and fetal outcome. Nutr. Rev.
64 (1), 15–30.
Skröder, H.M., Hamadani, J.D., Tofail, F., Persson, L.Å., Vahter, M.E., Kippler, M.J., 2014.
Selenium status in pregnancy influences children's cognitive function at 1.5 years of
age. Clin. Nutr. 34 (5), 923–930. http://dx.doi.org/10.1016/j.clnu.2014.09.020.
SRNT Subcommittee on Biochemical Verification, 2002. Biochemical verification of to-
bacco use and cessation. Nicotine Tob. Res. 4 (2), 149–159.
Stragierowicz, J., Mikołajewska, K., Zawadzka-Stolarz, M., Polańska, K., Ligocka, D.,
2013. Estimation of cutoff values of cotinine in urine and saliva for pregnant women
in Poland. Biomed. Res. Int. 2013, 386784. http://dx.doi.org/10.1155/2013/
386784.
Tamura, T., Goldenberg, R.L., Ramey, S.L., Nelson, K.G., Chapman, V.R., 2003. Effect of
zinc supplementation of pregnant women on the mental and psychomotor develop-
ment of their children at 5 y of age. Am. J. Clin. Nutr. 77 (6), 1512–1516.
Thompson, R.A., Nelson, C.A., 2001. Developmental science and the media: early brain
development. Am. Psychol. 56 (1), 5–15.
Turski, M.L., Thiele, D.J., 2009. New roles for copper metabolism in cell proliferation,
signaling, and disease. J. Biol. Chem. 284 (2), 717–721. http://dx.doi.org/10.1074/
jbc.R800055200.
Uriu-Adams, J.Y., Scherr, R.E., Lanoue, L., Keen, C.L., 2010. Influence of copper on early
development: prenatal and postnatal considerations. Biofactors 36 (2), 136–152.
http://dx.doi.org/10.1002/biof.85.
Wasowicz, W., Wolkanin, P., Bednarski, M., Gromadzinska, J., Sklodowska, M.,
Grzybowska, K., 1993. Plasma trace element (Se, Zn, Cu) concentrations in maternal
and umbilical cord blood in Poland. Relation with birth weight, gestational age, and
parity. Biol. Trace Elem. Res. 38 (2), 205–215.
Wasowicz, W., Gromadzinska, J., Rydzynski, K., Tomczak, J., 2003. Selenium status of
low-selenium area residents: Polish experience. Toxicol. Lett. 137 (1-2), 95–101.
Wehby, G.L., Murray, J.C., 2008. The effects of prenatal use of folic acid and other dietary
supplements on early child development. Matern. Child Health J. 12 (2), 180–187.
Yang, X., Yu, X., Fu, H., Li, L., Ren, T., 2013. Different levels of prenatal zinc and selenium
had different effects on neonatal neurobehavioral development. Neurotoxicology 37,
35–39. http://dx.doi.org/10.1016/j.neuro.2013.04.001.
Zachara, B.A., Wasowicz, W., Gromadzinska, J., Sklodowska, M., Krasomski, G., 1986.
Glutathione peroxidase activity, selenium and lipid peroxides concentrations in blood
from a healthy Polish population. Maternal and cord blood. Biol. Trace Elem. Res. 10
(3), 175–187. http://dx.doi.org/10.1007/BF02795616.
Zhu, L., Ji, X.J., Wang, H.D., Pan, H., Chen, M., Lu, T.J., 2012. Zinc neurotoxicity to
hippocampal neurons in vitro induces ubiquitin conjugation that requires p38 acti-
vation. Brain Res. 1438, 1–7. http://dx.doi.org/10.1016/j.brainres.2011.12.031.