Proteomics was used to identify a protein encoded by ORF 3a in a SARS-associated coronavirus (SARS-CoV). Immuno-blotting revealed that interchain disulfide bonds might be formed between this protein and the spike protein. ELISA indicated that sera from SARS patients have significant positive reactions with synthesized peptides derived from the 3a protein. These results are concordant with that of a spike protein-derived peptide. A tendency exists for co-mutation between the 3a protein and the spike protein of SARS-CoV isolates, suggesting that the function of the 3a protein correlates with the spike protein. Taken together, the 3a protein might be tightly correlated to the spike protein in the SARS-CoV functions. The 3a protein may serve as a new clinical marker or drug target for SARS treatment.
To better understand the mechanism underlying the hepatocellular carcinoma (HCC) metastasis and to search potential markers for HCC prognosis, differential proteomic analysis on two well-established HCC cell strains with high and low metastatic potentials, MHCC97-H and MHCC97-L, was conducted using two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/time-of-flight mass spectrometry. Cytokeratin 19 (CK19) was identified and found to be overexpressed in MHCC97-H as compared with MHCC97-L. This result was further confirmed by twodimensional Western blot analysis and immunofluorescence assay. Furthermore, one-dimensional Western blot analysis showed consistently increased CK19 expression in progressively more metastatic cells. Immunohistochemical study on 102 human HCC specimens revealed that more patients in the CK19-positive group had overt intrahepatic metastases (satellite nodules, p < 0.05; vascular tumor emboli, p < 0.001; tumor node metastatis staging, p < 0.001). For a better insight into the mechanisms of HCC metastasis, a high metastatic human HCC cell line, MHCC97, its clonal cells, MHCC97-H and MHCC97-L, with high and low metastatic potentials, and progressively more metastatic cells from lung metastatic lesions were established via repeated in vivo selection (3-5). These cells provide appropriate model systems with similar genetic background for the comparative study on the molecular events involved in HCC metastasis.Among the currently available techniques, proteomics permits the analysis of thousands of modified or unmodified proteins simultaneously and becomes increasingly popular for identifying biomarkers for early detection, classification, and prognosis of tumors, as well as pinpointing targets for improved treatment outcomes (6). We had used this approach to identify differentially expressed proteins between human HCC and normal liver cell line and investigated malignant growthassociated proteins in human hepatoma cells transfected with antisense-epidermal growth factor receptor (7,8). In this study, this technique was combined with immunology methods to study MHCC97-H and MHCC97-L for screening metastasis-associated proteins.Cytokeratins represent important structural components of the epithelial cytoskeleton, and their expression is remarkably tissue specific, suggesting that the type of cytokeratins present in the cells is related to their biological function (9). Recent
Carboxymethylcellulose (CMC) gel was used for encapsulating anionic herbicide 2,4-D for its controlled release. The slower release rate of 2,4-D in water and soil was obtained by adding in the gel formulation some modified bentonites, which were prepared by intercalating inorganic or organic cations in interlayers of Na(+)-saturated bentonite. The higher encapsulation efficiencies of 2,4-D were observed on the bentonite/CMC gel formulations due to the sorption of 2,4-D on modified bentonites. The time taken for 50% of 2,4-D to be released in water, t(50), showed a variation (8.8-19.8 h) for these gel formulations, the largest value corresponding to the formulation incorporated with hydroxy-iron intercalated bentonite that has the highest sorption capacity to 2,4-D. The gel formulations could also control the release of 2,4-D when applied to a thin soil layer and demonstrated potential to reduce leaching of such anionic herbicides in soil.
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