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Genomic DNA was extracted from whole blood samples, utilizing the salting out method.
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Subjects were required to submit overnight fasting blood samples utilized for the different metabolic parameters.
For blood samples, we utilized the tool Cn.
The collected blood samples were utilized immediately for the preparation of thin blood smears and were then kept at −20°C until DNA extraction.
While in those with regular menstrual cycles, blood samples were obtained during the follicular phase of the cycle, except for samples utilized for the assay of progesterone that performed in the day 21 of the cycle (luteal phase).
Their blood samples were utilized for ICC as confocal control specimens but not were used for FCM.
IGRAs differ from each other mainly with respect to the technique of IFN-γ detection (enzyme linked immunospot; ELISPOT vs. enzyme linked immunosorbent assay; ELISA) and the samples utilized (peripheral blood mononuclear cells vs. whole blood).
Therefore, this analysis utilized 40 blood samples collected in the morning prior to the start of a work-shift (pre-shift) and 38 blood samples were collected in the afternoon after all work activities had been completed (post-shift).
Another limitation is that we measured the biomarkers such as PS-PLA1 and glycerol LPLs in the blood samples taken from the arterial sheaths, while the venous blood samples are ordinarily utilized for the similar clinical studies.
This study utilized a low number of blood samples, and the sampling was mainly focused around calving and limited to only one herd.
Only endogenous 25(OH)D2 and/or 25(OH)D3 contained in human blood samples should be utilized for this purpose.
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