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Frozen cord samples were cryostat-sectioned (15 µm thick) in a sagittal plane at Kalderon's laboratory; the serially collected sections were thionin stained and examined by light microscopy.
Human post-mortem paraffin-embedded and frozen brain material, and frozen spinal cord samples were obtained from the Netherlands Brain Bank (NBB), Amsterdam.
FFPE brain and spinal cord samples were deparaffinized with xylene and DNA extracted in a manner similar to frozen samples after treatment with 6 mg Proteinase K for 3 days at 55°C.
Additionally, seven MS spinal cord samples were tested.
The spinal cord samples were dissected and stored as described above.
Spinal cord samples were collected for gene expression analysis from a total of 52 animals.
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The mechanism(s) involved in lead's impact on genomic DNA methylation levels in cord samples are unclear.
The amount of total protein in the spinal cord samples was measured with the PCA protein assay (PIERCE, USA).
The difference between serum anti-GAG antibody levels of controls and umbilical cord samples was significant (three-way ANOVA, F = 110.70; DF = 1.77; p < 0.001).
Frozen plasma from 210 cord samples was transferred on dry ice to the Centers for Disease Control and Prevention for laboratory analyses for the PBDEs and PBB-153.
The success of pyrosequencing in the umbilical cord samples was 100%, 97%, 99%, and 91% for Alu, LINE-1, p16, and p53, respectively.
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