Impact of Food Matrices on Digestibility of Allergens and Poorly Allergenic Homologs.

Background: Protease resistance is considered a risk factor for allergenicity of proteins, although the correlation is low. It is nonetheless a part of the weight-of-evidence approach, proposed by Codex, for assessing the allergenicity risk of novel food proteins. Susceptibility of proteins to pepsin is commonly tested with purified protein in solution. Objective: Food proteins are rarely consumed in purified form. Our aim was to evaluate the impact of experimental and endogenous food matrices on protease susceptibility of homologous protein pairs with different degrees of allergenicity. Methods: Porcine and shrimp tropomyosin (ST) were subjected to sequential exposure to amylase, pepsin, and pancreatin in their respective endogenous matrix (pork tenderloin/boiled shrimp) and in three different experimental matrices (dessert mousse [DM], soy milk [SM], and chocolate bar [CB]). Digestion was monitored by immunoblotting using tropomyosin-specific antibodies. Recombinant peach and strawberry lipid transfer protein were biotinylated, spiked into both peach and strawberry fruit pulp, and subjected to the same sequential digestion protocol. Digestion was monitored by immunoblotting using streptavidin for detection. Results: Chocolate bar, and to a lesser extent SM, had a clear protective effect against pepsin digestion of porcine tropomyosin (PT) and to a lesser extent of ST. Increased resistance was associated with increased protein content. Spiking experiments with bovine serum albumin (BSA) confirmed the protective effect of a protein-rich matrix. The two tropomyosins were both highly resistant to pepsin in their protein-rich and lean native food matrix. Pancreatin digestion remained rapid and complete, independent of the matrix. The fat-rich environment did not transfer protection against pepsin digestion. Spiking of recombinant peach and strawberry lipid transfer proteins into peach and strawberry pulp did not reveal any differential protective effect that could explain differences in allergenicity of both fruits. Conclusions: Protein-rich food matrices delay pepsin digestion by saturating the protease. This effect is most apparent for proteins that are highly pepsin susceptible in solution. The inclusion of food matrices does not help in understanding why some proteins are strong primary sensitizers while homologs are very poor allergens. Although for induction of symptoms in food allergic patients (elicitation), a protein-rich food matrix that may contribute to increased risk, our results indicate that the inclusion of food matrices in the weight-of-evidence approach for estimating the potential risks of novel proteins to become allergens (sensitization), is most likely of very limited value.

The N-terminal 303 amino acids of the human cytomegalovirus envelope glycoprotein B (UL55) and the exon 4 region of the major immediate early protein 1 (UL123) induce a cytotoxic T-cell response.

We reported earlier that an adenovirus (Ad) recombinant expressing the full-length human cytomegalovirus (HCMV) glycoprotein B (gB) gene induces gB-specific cytotoxic T lymphocyte (CTL) responses in CBA (H-2k) mice (Berencsi et al., J. Gen. Virol. 74, 257-2512, 1993). Here we show that mice immunized with Ad recombinant viruses expressing truncated forms of the gB gene containing the first 700 (Ad-700), 465 (Ad-465) or 303 (Ad-303) amino acids of gB or an Ad construct containing exon 4 (E4) of the HCMV immediate early 1 (IEI) gene (Ad-IEI (E4)) demonstrate HCMV-specific CTL responses. These data suggest the importance of the first 303 amino acids of the gB polypeptide and the IEI E4 product in designing a vaccine to induce anti-HCMV CTL responses.

Regulatory oversight of biochemical pesticides by the U.S. Environmental Protection Agency: health effects considerations.

The Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) enables the Office of Pesticide Programs (OPP) of the U.S. Environmental Protection Agency to ensure that pesticide use in commerce will not result in unreasonable adverse effects to humans and the environment. Currently, two classes of pesticides are recognized: conventional chemical pesticides and biological pesticides. The latter group is divided into biochemical and microbial pesticides. The recent resurgence of biochemical pesticides as effective pest control agents has increased the number of applications for experimental use permits and for product registration. The fundamental information and data necessary to evaluate such products by the Health Effects Division (HED) of OPP are discussed, as well as the criteria for the classification of a pesticide as a biochemical versus a conventional chemical pesticide. In accordance with the Agency's effort to encourage the development of pesticides less toxic to humans and the environment, the scientific basis for providing future regulatory relief and reduced data requirements for biochemical pesticides is discussed.

DEPRESSED METABOLIC ACTIVITY OF VESICULAR-ARBUSCULAR MYCORRHIZAL FUNGI AFTER FUNGICIDE APPLICATIONS.

Application to soil of either benomyl or captan significantly decreased the growth of vesicular-arbuscular mycorrhizal (VAM) onion (Allium cepa L. cv. Hyper) plants 4 weeks after treatment but non-VAM plants were not affected. Fungal colonization of the onion roots, as indicated by non-vital staining with chlorozole black E, was depressed 2 weeks after fungicide application. However, decreases in metabolically active VAM fungal tissue, revealed by a succinate dehydrogenase assay, could be detected as soon as 3 d after fungicide treatment. There was little difference between the fungicides in their effect on the VAM fungi used. The usefulness of the succinate dehydrogenase assay in predicting effects of fungicide is discussed.