We conclude that divergent allergenicity of apple strains mainly depends on different expression levels of the major allergen. Introduction of a proline residue in position 111 of Mal d 1 and in position 112 of Bet v 1 led to a drastic reduction of allergenicity of both the pollen and the food allergen, obviously also removing the cross-reactive epitope. Mutants with reduced IgE-reactivity but maintained T-cell reactivity may represent new candidates for a safer specific immunotherapy with reduced side-effects.

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Serum testing of genetically modified soybeans with special emphasis on potential allergenicity of the heterologous protein CP4 EPSPS" (2007). Faculty Publications in Food Science and Technology. 156.

Mal d 1, the major apple allergen shows strong cross-reactivity with IgE specific for the major birch pollen allergen, Bet v 1, and is responsible for birch pollen related food allergy to apple. In contrast to other food and pollen allergens, only a few data on the B-cell epitopes of Mal d 1 are available. To obtain data on the antibody binding epitopes of Mal d 1 and to gain insights to the structures responsible for its B-cell cross-reactivity to Bet v 1, the binding characteristics were studied with 3 monoclonal antibodies (MAbs) raised against Mal d 1 and with patients' IgE directed against Mal d 1 and Bet v 1 by immunoblotting and ELISA. The different binding characteristics of these three MAbs indicated that the MAbs 1D6, 2B2 and 3F8 recognized different epitopes on the major apple allergen. MAb 2B2 cross-reacted with Bet v 1 on immunoblots, whereas 1D6 and 3F8 did not. Since 1D6 and 2B2 (but not 3F8) always competed with IgE from the same apple-allergic patients, 1D6 may be a useful tool for Mal d 1 epitope studies and 2B2 for an epitope that cross-reacts with IgE specific for Bet v 1.

The biological activities of Smilax china L. rhizome (SCR), hot water (SCRW) and 70% ethanol extract (SCRE) were analyzed. The total phenolic contents of SCRW and SCRE were 51.7 and 100.5 mg/g, respectively. The measured flavonoid content of SCRW (61.7 μg/g) was almost double that of SCRE (31.7 μg/g). SCRE (IC50=42.4 μg/mL) exhibited stronger antioxidant activity in the DPPH system than the positive control α-tocopherol (71.3 μg/mL) or butylated hydroxy anisole (53.8 μg/mL) did. SCRE (IC50=50.3 μg/mL) also showed stronger ABTS radical scavenging activity, as did α-tocopherol (67.1 μg/mL). The SOD-like activity and Tyrosinase inhibition activity of SCRW and SCRE showed almost the same pattern. The best SOD-like activity and tyrosinase inhibition activity were measured as 24.9% and 20.3% in SCRW at 1,000 μg/mL, respectively. The cytotoxic effects of the SCR extracts were analyzed via MTT assay on human cancer and normal cells. SCRW and SCRE did not show cytotoxicity up to the concentration of 1,000 µg/mL against the normal human cell line HEK293. Against human breast cancer cells (MCF-7), SCRW inhibited MCF-7 growth (by 27.6%) better than the anticancer drug cyclophosphamide (15.5%) at 1,000 μg/mL. SCRE (1,000 µg/mL) inhibited the growth of human lung cancer cells A549 (37.6%) and human stomach cancer cells AGS (53.6%) more effective than did SCRW (21.0% and 35.4%) or CPA (22.2% and 31.7%). These results suggest the potential use of SCRE and SCRW as an excellent antioxidant and antiproliferative substance, respectively. Key words：anticancer, antioxidant, 1,1-diphenyl-2-picrylhydrazyl, polyphenol, Smilax china L.