SummaryThe IgE-binding proteins in soybeans were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the frac tionated soybean proteins probed with the sera of the patients with atopic dermatitis. About 20% of the patients examined were shown to have specific IgE antibodies to soybean proteins. At least 16 soybean proteins with molecular weights ranging from about 70,000 to 14,000 were rec ognized by the sera of the patients: 10 major IgE-binding components were found in the 7S-globulin fraction, and the others mainly in the 2S-globulin and whey fractions. The IgE antibodies of the patients bound most strongly and frequently to a unique protein with molecular weight of about 30,000 in the 7S-globulin fraction, which appeared to be the major allergen in soybeans and was named as Gly m Bd 30K. The proteins in the 11 1S-globulin fraction were scarcely recognized by the patients' sera and assumed to be less allergenic for the patients with atopic dermatitis.

The soybean allergenic protein, Gly m Bd 30K [Ogawa et al., J. Nutr. Sci. Vitaminol., 37, 555-565 (1991)] which is most strongly and frequently recognized by the IgE antibodies in sera of soybean-sensitive patients with atopic dermatitis, has been characterized. The allergen was isolated from the crude 7S-globulin fraction as an oligomeric form with a molecular weight of more than 3000,000 by gel-filtration chromatography. On two-dimensional gel electrophoresis, the native oligomeric allergen had an isoelectric point of about pH 4.5 and was dissociated into a monomeric form with a molecular weight of about 32,000 by the treatment with sodium dodecyl sulfate and 2-mercaptoethanol. The monomeric allergen had an N-terminal amino acid sequence and amino acid composition identical with those of the soybean seed 34-kDa oil-body-associated protein or the soybean vacuolar protein P34 with close homology to papain-like thiol proteinases [Kalinski et al., J. Biol. Chem., 267, 12068 (1992)]. The identity was further confirmed by the immunological cross-reactivity to the antibodies produced against each of the purified allargen and the 34-kDa oil-body-associated protein. By this observation, Gly m Bd 30K was shown to have about 30% sequence homology with Der pI, a house dust mite allergen that is a thiol proteinase from Dermatophagoides pteronyssius.

SummaryWe investigated the association between nutrient biomarkers and dietary intake estimated using a brief self-administered dietary history questionnaire (BDHQ) for Japanese children and adolescents. Blood samples were collected from 398 subjects (5th graders of elementary school aged 10-11 y, and 2nd graders of secondary schools aged 13-14 y) randomly selected from among students in Shunan City, Japan, who were then required to answer two questionnaires. Spearman correlations were calculated between dietary intake and the corresponding biomarkers (serum carotenoids, tocopherols, and erythrocyte fatty acids). Correlations with ␤ -carotene and ␤ -cryptoxanthin were significant in the 13-and 14-y age group ( r ϭ 0.220-0.333, p Ͻ 0.030) and the 10-and 11-y age subgroup who answered the questionnaire with assistance ( r ϭ 0.295-0.299, respectively, p ϭ 0.006). Consumption of green-yellow vegetables and fruits was significantly correlated with ␤ -carotene and ␤ -cryptoxanthin levels ( r ϭ 0.205-0.341, p Ͻ 0.047). In the 13-and 14-y age group, correlations with eicosapentaenoic and docosahexaenoic acids were between 0.215 and 0.473 ( p Ͻ 0.040). Total seafood intake was significantly correlated with marine n -3 polyunsaturated fatty acids (PUFAs; r ϭ 0.239-0.420, p Ͻ 0.023). In the 10-and 11-y age subgroup who completed the questionnaire with assistance, seafood intake was significantly correlated with marine n -3 PUFAs ( r ϭ 0.239-0.243, p Ͻ 0.032). In conclusion, dietary intake assessed using the BDHQ reflects the corresponding biomarkers for 13-and 14-y-olds; however, when used for elementary school children, caution is necessary in interpreting the results.

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The soy protein-galactomannan conjugate prepared by the Maillard reaction removed the allergenicity of the 34 kDa protein which is frequently recognized by the IgE antibody in the sera of soybean-sensitive patients as a major allergen. Monitoring of polyclonal antibody titers by an indirect enzyme-linked immunosorbent assay and immunoblotting of rabbit sera, monoclonal antibody, and human allergic sera showed that soy protein-galactomannan conjugation was more effective in reducing the allergenicity of the soy protein than transglutaminase treatments and/or chymotrypsin. The soy protein-galactomannan conjugate was highly soluble at all pHs, while untreated soy protein was sparingly soluble at pH 4-6. Heat stability and emulsifying properties were greatly improved by conjugation with galactomannan.

Carotenoids are known to be potent quenchers of singlet molecular oxygen [O2 (1Δg)]. Solar light-induced photooxidative stress causes skin photoaging by accelerating the generation of reactive oxygen species via photodynamic actions in which O2 (1Δg) can be generated by energy transfer from excited sensitizers. Thus, dietary carotenoids seem to participate in the prevention of photooxidative stress by accumulating as antioxidants in the skin. An in vivo study using hairless mice clarified that a O2 (1Δg) oxygenation-specific peroxidation product of cholesterol, cholesterol 5α-hydroperoxide, accumulates in skin lipids due to ultraviolet-A exposure. Matrix metalloproteinase-9, a metalloproteinase family enzyme responsible for the formation of wrinkles and sagging, was enhanced in the skin of ultraviolet-A -irradiated hairless mice. The activation of metalloproteinase-9 and the accumulation of 5α-hydroperoxide, as well as formation of wrinkles and sagging, were lowered in mice fed a β-carotene diet. These results strongly suggest that dietary β-carotene prevents the expression of metalloproteinase-9 (at least in part), by inhibiting the photodynamic action involving the formation of 5α-hydroperoxide in the skin. Intake of β-Carotene therefore appears to be helpful in slowing down ultraviolet-A -induced photoaging in human skin by acting as a O2 (1Δg) quencher.

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The mechanism of immunological benefits induced by carotenoids has not been fully elucidated. Here, we investigated some of the immunity-related properties of beta-carotene and two other carotenoids, beta-cryptoxanthin, and lutein, on the murine macrophages cell line RAW264. beta-Carotene added to the culture medium accumulated in the cells in a time- and dose-dependent manner. The accumulation was positively correlated with cellular lipid peroxidation, demonstrating the pro-oxidative activity of beta-carotene, and also with the synthesis of glutathione, an intracellular antioxidant. Conversely, accumulation of beta-carotene was negatively correlated with the transcription of immune-active molecules, such as IL-1beta, IL-6, and IL-12 p40, in cells stimulated by LPS and INF-gamma. The transcription of the pro-inflammatory cytokines IL-1beta and IL-6 was more sensitive to the accumulation of beta-carotene than was IL-12 p40. The accumulation of beta-cryptoxanthin in cells resulted in effects similar to those of beta-carotene. However, lutein accumulated minimally and did not significantly affect the cells. These results demonstrate that beta-carotene, and beta-cryptoxanthin as well, can accumulate in RAW264 cells and induce changes in intracellular redox status, which in turn regulate the immune function of macrophages.