Development of a monoclonal antibody-based indirect competitive immunosorbent assay for 4(5)-Methylimidazole detection in caramels.

In this study, an indirect competitive enzyme-linked immunoassay (ic-ELISA) based on monoclonal antibody for 4(5)-Methylimidazole (4-MI) detection was described. The artificial antigens were prepared by conjugating bovine serum albumin (BSA) or ovalbumin (OVA) with the hapten of 4-MI. And monoclonal antibody, evaluated by ic-ELISA, was obtained by immunizing BABL/c mice. After optimizing, a standard curve for ic-ELISA detection on 4-MI was obtained with the linear detection range of 0.64-20.48 mg/L. The cross-reactivity (CR) of all the structural analogues of 4-MI was less than 5.62%. The recoveries of 4-MI in caramels detection were ranged from 88.69% to 114.09%, with relative standard deviation (n=3) below 8.07%. The results suggested that the established ic-ELISA is promising for 4-MI commercial detection in caramels.

[Apoptosis of lens epithelial cell induced by curcumin and its mechanism].

OBJECTIVE: To investigate the effects of natural drug curcumin (Cur) on apoptosis of lens epithelial cell (LEC) in vitro and its mechanism. METHODS: The bovine LEC were cultured with Cur, the ultrastructure changes were observed under transmission electron microscope (TEM), the DNA content and mitochondrial transmembrane potential (DeltaPsim) changes were studied by flow cytometry (FCM). RESULTS: The typical morphological changes of LEC apoptosis in Cur group detected by TEM included chromatin condensation and aggregation at the periphery of the nucleons and nuclear fragmentation. The DNA content of LEC in Cur group decreased time-dependently. The DNA content was significantly lower than that of the control group (P < 0.01). The DeltaPsim of LEC in Cur group was decreased, appeared in early stage (8 hours) and reached the maximum after 72 hours. The difference of DeltaPsim of LEC between Cur group and the control group was significant (P < 0.01). CONCLUSIONS: Cur can remarkably induce apoptosis of LEC in vitro. Cur induced LEC apoptosis is caused by decrease of DNA content in LEC nucleus. Collapse of DeltaPsim in cytoplasm induced by Cur results in the irreversible apoptosis process of LEC. This is the early event of LEC apoptosis. LEC apoptosis induced by Cur may pass through two pathways: nuclear pathway and cytoplasmic pathway. The apoptosis of LEC induced by Cur may be the cellular and molecular mechanisms of reducing lens posterior capsular opacification by Cur. Cur may become an effective and low toxic medication for the prevention and treatment of after-cataract.