Immunoreactivities of p11 and calcyclin in baker's yeast, wheat germ and lobster.

After effectively eliminating the nonspecific cross-immunoreactivity with the affinity columns of anti-IgG agarose and IgG agarose, the potent immunoreactivities of p11 and calcyclin in wheat germ, lobster tail muscle, and three strains of baker's yeast were analyzed by Western blotting using mouse anti-p11 and rabbit anti-calcyclin. The occurrence of multiple bands may be due to either autolyses and/or the interactions between the p11 (or calcyclin) and other endogenous biological molecules. The results suggest not only a ubiquitous distribution and a universal Ca(2+)-mediating regulatory role of p11 and calcyclin in eukaryotes, but also an evolutionary conservation of these (S-100)-related proteins.

Interaction of polyadenylic acid (5') with histone H1 or protamine.

The interaction between polyadenylic acid (5') (poly [A]) and histone (or protamine) was analyzed by electrophoretic retardation of poly [A]-histone (or protamine) complex in agarose gel. The potency of interaction was protamine > histone H1, arginine-rich histone > other histones. The catalytic subunit of cyclic AMP-dependent protein kinase effectively decreased the electrophoretic retardation of poly [A]-histone H1. The interaction between poly [A] and histone H1 was also detected by the drastically enhanced absorbance around 340 nm. The findings may implicate a regulatory role of histone H1 on mRNAs through its binding on poly [A] tails.

Immunoreactivities of m-calpain, calpastatin, nitric oxide synthase, myelin basic protein and dynamin II in baker's yeast, wheat germ and lobster tail muscle.

Vertebrate m-calpain, calpastatin, constitutive nitric oxide synthase, myelin basic protein, and dynamin I are substrates of protein kinase C (PKC). The presence/absence of similar/related protein in nonvertebrate was investigated by immunological methods, including (1) affinity chromatography on agarose-secondary antibodies and agarose IgG for removal of nonspecific immunoreactivities from crude extracts; (2) omitting beta-mercaptoethanol treatment and boiling prior to SDS-PAGE to increase the immunoreactivity; (3) immunoreactivity comparisons of nonspecific IgG as controls with specific anti-(vertebrate PKC-substrates/related proteins) in Western blots. It was found that (a) m-calpain and dynamin I were absent in baker's yeast, wheat germ and lobster tail muscle, (b) m-calpain, nitric oxide synthase, myelin basic protein and dynamin II were present in all three samples, and (c) calpastatin was present in baker's yeast and lobster tail muscle. The presence and absence of these proteins suggest evolutionary conservation and divergence, respectively, of these PKC substrates.

Predominantly indurated reactions to sensitizers may not cause keratinocytes to express HLA-DR.

Dermal reactions to primary intradermal or appendageal sensitization are compared to predominantly dermal reactions to standard patch tests and to intradermal antigen tests. In contrast to epicutaneous spongiotic contact dermatitis, HLA-DR was only seen on skin appendages and nearby basal keratinocytes in indurated tissue reactions with the exception of the reactions with focal basal cell layer disruption and an indurated patch test performed one week post angry back syndrome. Other intradermal skin tests showed only minimal epidermal HLA-DR expression despite spongiotic epidermal changes. Predominantly dermal hypersensitivity reactions can be induced by intradermal or epicutaneous routes. They can evoke hypersensitivity responses which do not cause most epidermal keratinocytes to express HLA-DR.