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Finally, wells were washed thrice with PBS and C4bp binding was detected colorimetrically by using tetramethylbenzidine substrate (Thermo Fisher Scientific) and reading at 450 nm using the HIDEX Sense plate reader

MTT viability assays were performed as described elsewhere (43)

Therefore, even considering the burdens for medical professionals and medical economy, direct observation of vocal cord movements before surgery is recommended for all surgical cases

(1): Hyperpigmentation over tongue, bilateral hands and feet Diagnosis Clinical neurological examination was suggestive of non-compressive myelopathy