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On the day of the assay, plates were removed from the incubator and each well was washed twice with 100 l HBSS (Gibco, Life Technologies) and pre-incubated for 30 min at 37 C with 85 l HBSS per well

Elsevier Masson

The culprit might be hormonal imbalance and weight gain

The autoantibodies in this process against the TSH receptor continuously stimulate the thyroid gland toward hyperplasia and production of increased amounts of hormones (Antonelli et al