A Comparison between Three Different Automated Total 25-hydroxyvitamin D Immunoassay Methods and Ultra Performance Liquid Chromatography.

OBJECTIVE: Vitamin D is a fat soluble vitamin responsible for calcium metabolism and more recently discovered effects. This led to an increase in requests for vitamin D test by clinicians. New automated assays have been introduced for 25-hydroxyvitamin D measurement. METHODS: Results from these new method have to be related to a Standard method to obtain best results for practical usage. In our study, one hundred venous blood samples were analyzed for 25-OH vitamin D on three immunological methods in our lab and correlated with ultra-performance liquid chromatography (UPLC) method as a reference method. RESULTS: Statistically analysis of results obtained for correlations between the 3 methods against the reference UPLC was done by Spearman's Correlation. It showed positive correlation in all methods with significant p value < 0.001. Differences and biases between methods were evaluated using a Bland-Altman plot and Cohen's Kappa agreement. Best agreement was found in Cobas 6000 followed by the Access2 then comes Architect.  Conclusions: All immunoassays can be used in routine 25(OH) D measurements, still some methods are better than others. A clinical laboratory must at least be aware of its method to avoid misinterpretation of results.

Ezrin expression is altered in mice lymphatic metastatic hepatocellular carcinoma and subcellular fractions upon Annexin 7 modulation in-vitro.

Ezrin and Annexin seven (A7) have been suggested to be involved in several roles in cancers metastasis. However, the role of Ezrin and the effect of A7 on Ezrin expression in lymphatic metastatic hepatocellular carcinoma (LNM-HCC) have not been extensively explored yet. This study reports expression of Ezrin in high lymphatic metastasis (Hca-F >70%) and low metastatic metastasis (Hca-P <30%) HCC cell lines, and the effect of A7 on Ezrin expression. Real-Time PCR, Western blot, Subcellular fractionation, Immunocytochemistry and Immunofluorescence were used to investigate Ezrin expression in addition to migration and invasion behaviors of A7 up-regulated Hca-F cells, A7 down-regulated Hca-P and in their respective negative control (NC) cells. Ezrin expression was higher in high LNM-HCC than low LNM-HCC (p=0.0046). Cell fractionation analysis reveals that Ezrin was highly present in the cytoplasm, nucleus and cytoskeleton of NC-Hca-F cells. However, Ezrin was highly observed in the cell membrane, nucleus and cytoskeleton of NC-Hca-P cells. A7 up-regulation in Hca-F suppressed Ezrin expression (p=0.0248), but increase the migration and invasion, whereas Ezrin was mainly located in the cytoplasm and nucleus fractions. Down-regulation of A7 in Hca-P cells, enhanced Ezrin expression (p<0.0001) in the cytoplasm and nucleus fractions, and suppressed migration and invasion. In conclusion, Ezrin may play a role in LNM-HCC and might be inversely associated with A7 expression. The subcellular localization of Ezrin and A7 was varied according to the metastatic levels. Ezrin may thus be a potential diagnostic and/or prognostic biomarker for HCC.