Oligosaccharide chains of IgG purified from sera of patients with rheumatoid arthritis (RA) and various cancers were investigated with fluorophore-assisted carbohydrate electrophoresis (FACE). In addition, the relationship between the increased fraction separated by the FACE and the production of anti-agalactosyl IgG antibody was studied. The fluorescence-labeled sugar chains were separated into five fractions (Fr1-5) by PAGE. The Fr4, oligosaccharide lacking galactose, revealed significantly higher levels in RA and cancer (p<0.01), while the Fr1, oligosaccharide with galactose, was significantly decreased in both pathological conditions (p<0.01). Therefore, it suggests that augmentation of the abnormal IgG in sera was not specific to patients with RA. In patients with cancer, the Fr3, oligosaccharide with mono-sialic acid, showed an increased tendency associated with significant increase of the Fr5, oligosaccharide with di-sialic acid (p<0.01), while in patients with RA the Fr3 was found to have a decreased tendency. There was a significant correlation between the contents of Fr4 and anti-agalactosyl antibodies, an autoantibody against galactose free IgG (p<0.01). In patients with either RA or cancer, the Fr4 increasing above apporoximately 30% was associated with abnormally high level of the anti-agalactosyl antibodies. Accordingly, this finding suggests that the increased Fr4 was related to the production of the anti-agalactosyl IgG antibodies.

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The expression level of guard hair fiber (following hair fiber) protein in the Korean native goat showed remarkably seasonally change. The protein components (M.W.: 9kDa) specific for the cashmere fiber were isolated. To determine the N-terminal amino acid sequences on five kinds of major proteins for hair fiber and cashmere fiber, protein components (50 and 42kDa) of the hair fiber which showed seasonal change in the level of protein expression were determined as T-L-G-F-Y-T-A-G-P-A-F-L-L-V---F-Q-K and I-L-N-G-T-R-T-R-P-F-R-F-I-I-P, respectively. And this 42kDa protein component had the homology (61.5%) with integrase. A common protein (17kDa) having sequence of T-G-S-C-C-G-P-T-F between the hair fiber and the cashmere fiber was identified as a keratin protein. N-terminal amino acid sequences of cashmere fiber proteins with 50kDa and 42kDa, were determined as T-L-G-F-Y-T-A-G-P-A-F-L-L-V and I-L-N-G-T-------P-F, respectively, and the sequence of 17kDa keratin protein was determined as T-G-S-C-C-G-P-T-F-S. Also that of 9kDa, which is specific for the cashmere fiber protein was determined as T-G-F-T-(G)-G-G-I-Y-F-P-G---T, which might be the novel protein component.

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To objectively evaluate the relationship between psychiatric disorders and stress, we attempted to measure the levels of 70 kilodalton (kDa) heat shock proteins (HSP70) in peripheral blood mononuclear cells (PBMC) using ELISA. Although the standard curve created from measurements of a commercial HSP70 (from bovine brain; S-HSP70) showed good linearity, measurements of HSP70 in PBMC samples appeared negative when concentrations were calculated using this curve. We determined that addition of the detergent used for PBMC lysis to dilutions of S-HSP70 also resulted in negative calculations. Therefore, when we dissolved the PBMC in distilled water instead of the detergent, HSP70 levels could be correctly determined. To verify the accuracy of HSP70 calculations for the PBMC samples dissolved in distilled water, we compared the ELISA results with those obtained by SDS-polyacrylamide gel electrophoresis and Western blotting for PBMC dissolved in the detergent. Both methods resulted in similar patterns of HSP70 expression.

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