Trial Information

Polish Mother and Child Cohort Study (REPRO_PL).Follow up of the Children.

The Polish Mother and Child Cohort is multicentre prospective study on different exposures.

Prospective cohort study design enables identification of exposures that may influence
pregnancy outcome and chil-dren's health, verification of such exposures by biomarker
measurements and notification of any changes in exposure levels.

The aim of the study is to evaluate the impact of exposure to different environmental
factors during pregnancy and after birth on pregnancy outcome and children's health.
Specific research hypotheses refer to the role of heavy metals, exposure to polycyclic
aromatic hydrocar-bons (PAHs) and environmental tobacco smoke (ETS) in the aetiology of
intrauterine growth retardation (IUGR), preterm delivery (PD) and the risk of respiratory
diseases, allergy and poor mental and physical development. It is also intended to explain
the role of oxidative stress and nutritional status of the pregnant women. The impact of
occupational exposures and stressful situations on pregnancy outcome will be evaluated from
question-naire data.

The results of the study will help to determine levels of child prenatal and postnatal
exposure in several areas of Poland and their im-pact on course and outcome of pregnancy and
children's health.

This protocol concerns the children that are followed-up from birth to the age of 2 years to
deter-mine long term effects of pre- and postnatal environmental exposures.

Exposure of children to ETS. Postnatal children exposure to ETS at 12, and 24 months after
birth is assessed. From the children urine sample and saliva samples from mothers who will
declare smoking abstinence are collected. The saliva and urine cotinine level is analysed
using high performance liquid chromatography coupled with tandem mass spectrometry/positive
electrospray ionization (LC-ESI+MS/MS) and isotope dilution.

Exposure of children to PAH. The HPLC technique is used for the analysis of the level of
1-hydroksypyrene in urine as the biomarker of PAH exposure (12 and 24 months).

Exposure to lead, cadmium, mercury in children. Blood and hair are collected for about 100
children. Total and inorganic mercury is analyzed using cold vapour atomic fluorescence
spectrophotometry. The lead and cadmium concentrations in blood is determined by
graphite-furnance atomic-absorption spectrometry.

Exposure to indoor allergens, endotoxins and glucans. The analyzis is covered approx. 50
family homes of children. The settled dust is collected from floors in leaving rooms and in
children beds (mattresses and blankets). To determine allergens concentrations in dust
samples, the commercially available enzymelinked immunosorbent assey (ELISA) kits ispplied
according to the protocol.To determine endotoxins and (1→3)-β-D-glucans concentrations the
LAL test will be applied. The questionnaire study is performed among parents in order to
obtain the data about demographic factors and home conditions.

Questionnaires conducted with mothers 12, and 24 months after delivery. Mothers of all
children are interviewed. The questionnaires investigate the children's health and
development, the nutrition of the small child and their contacts with other children. The
smoking status of the parents and other potential sources of children's ETS exposure are
identified. Information about proximity to heavy traffic is inquired. Additionally the
information from the closets to his/her place of living air pollution monitoring station is
collected. Exposure to other environmental factors including domestic animals, home dust,
household cleaning agents and indoor used pesticides is evaluated by the questionnaire.

Determination of the prevalence of the upper and lower respiratory tract infections, middle
ear diseases and allergy among the child. The incidence of the upper and lower respiratory
tract infections, middle ear diseases and symptoms of allergy among the child is identified
based on the detailed interview with the mothers in 12 and 24 month after child birth. This
interviews is conducted by paediatricians. For each child upper and lower respiratory tract
infections (tonsillitis, laryngitis, bronchitis and pneumonia), as well as middle ear
diseases and allergy to food and inhalant allergens is identified based on the review of
their medical charts. The duration of each infection and disease, medications taken and
hospitalization is identified. The current health status of the chid and child'
anthropometry (high, weight) is preformed by pediatrician.

Determination of the neurodevelopment of the children. BSID-III is used for assessment of
child neurodevelopment. The tests is scheduled on 12 and/ 24 months of live and is conducted
by psychologist or child development specialist. The test assesses five developmental areas
of the child: cognitive, motor (fine and gross motor), language (receptive, expressive),
social-emotional and adaptive behavior.