Aeroallergens and pollinosis: Molecular and immunological characteristics of cloned pollen allergens

Affecting almost twenty percent of thepopulation in industrialized countries,allergic diseases such as asthma and seasonalrhinitis result from the development ofimmediate hypersensitivity to otherwiseinnocuous components of the environment. Pollen, the male gametophyte of flowering plantspecies, is one of the most predominant sourcesof environmental allergens, and a significantcause of allergic diseases. This reviewdiscusses the nature of pollen proteins asallergens, their effect on the human immunesystem and their mode of environmentaltransmission, including effects of theinteraction between pollen aeroallergens andair pollution. The influence of cross-reactivepollen allergens to the incidence of foodsensitivities is also discussed. Floweringplant species that possess allergenic pollen,identified with allergens cloned from thesespecies, are also discussed.     

Detection and side-chain specificity of IgE antibodies to flucloxicillin in allergic subjects

Unlike studies on the antigenicity of penicillins in laboratory animals, limited information is available on the allergenicity of penicillins in man, especially with regard to fine structural allergenic differences between the many different penicillins. Inconsistent with the earlier conclusions of others, our studies suggest that side-chain structures to flucloxacillin-reactive IgE antibodies. Quantitative hapten inhibition studies revealed potent inhibition by flucloxacillin and three structurally related penicillins: oxacillin, cloxacillin and dicloxacillin. Analysis of the results showed that the side-chain group of flucloxacillin, 3-2(-chloro-6-fluorophenyl)-5-methyl-4-isoxazolyl, is recognized by some antibodies and that the 5-methyl-3-phenyl-4-isoxazolyl group, with or without halogen substituents, accounts for the reactivity of other antibodies and for the cross-reactions seen with some other penicillins. Since it is the side-chain group that distinguishes the many different types of penicillin, and since IgE antibodies in many of the allergic reactions recognize the side-chain groups, it is becoming clear that specific assays are required for the detection of IgE antibodies to each of the different penicillins.     

Allergens from short ragweed leaf tissue cultures

Summary Short ragweed (Ambrosia elatior L.) leaf cells were grown as callus and suspension cultures. Immunodiffusion tests against anti-pollen crude extract and anti-antigen E sera did not detect antigen E in the tissue cultures. However, two allergenically active fractions were isolated from the tissue cultures by ammonium sulfate precipitation, diethylaminoethyl-cellulose and Sephadex G-100 chromatography. Two fractions isolated (Sephadex G-100, fractions II and III) were electrophoretically homogeneous, carried net charges similar to that of antigen E in polyacrylamide disc electrophoresis at pH 8.9 and had molecular weights of 10,000 to 15,000 and 30,000 to 40,000, respectively. These fractions were highly skin reactive in individuals sensitive to short ragweed pollen. This paper was abstracted from a thesis submitted by A. Shafiee to the University of Minnesota in partial fulfillment of the Doctor of Philosophy degree requirements.     

The diversity of arthropods in homes across the United States as determined by environmental DNA analyses

We spend most of our lives inside homes, surrounded by arthropods that impact our property as pests and our health as disease vectors and producers of sensitizing allergens. Despite their relevance to human health and well‐being, we know relatively little about the arthropods that exist in our homes and the factors structuring their diversity. As previous work has been limited in scale by the costs and time associated with collecting arthropods and the subsequent morphological identification, we used a DNA‐based method for investigating the arthropod diversity in homes via high‐throughput marker gene sequencing of home dust. Settled dust samples were collected by citizen scientists from both inside and outside more than 700 homes across the United States, yielding the first continental‐scale estimates of arthropod diversity associated with our residences. We were able to document food webs and previously unknown geographic distributions of diverse arthropods – from allergen producers to invasive species and nuisance pests. Home characteristics, including the presence of basements, home occupants and surrounding land use, were more useful than climate parameters in predicting arthropod diversity in homes. These noninvasive, scalable tools and resultant findings not only provide the first continental‐scale maps of household arthropod diversity, but our analyses also provide valuable baseline information on arthropod allergen exposures and the distributions of invasive pests inside homes.     

Tubers from potato lines expressing a tomato Kunitz protease inhibitor are substantially equivalent to parental and transgenic controls

Recombinant protease inhibitors represent useful tools for the development of insect‐resistant transgenic crops, but questions have been raised in recent years about the impact of these proteins on endogenous proteases and chemical composition of derived food products. In this study, we performed a detailed compositional analysis of tubers from potato lines expressing the broad‐spectrum inhibitor of Ser and Asp proteases, tomato cathepsin D inhibitor (SlCDI), to detect possible unintended effects on tuber composition. A compositional analysis of key nutrients and toxic chemicals was carried out with tubers of SlCDI‐expressing and control (comparator) lines, followed by a two‐dimensional gel electrophoresis (2‐DE) proteomic profiling of total and allergenic proteins to detect eventual effects at the proteome level. No significant differences were observed among control and SlCDI‐expressing lines for most chemicals assayed, in line with the very low abundance of SlCDI in tubers. Likewise, proteins detected after 2‐DE showed no quantitative variation among the lines, except for a few proteins in some control and test lines, independent of slcdi transgene expression. Components of the patatin storage protein complex and Kunitz protease inhibitors immunodetected after 2‐DE showed unaltered deposition patterns in SlCDI‐expressing lines, clearly suggesting a null impact of slcdi on the intrinsic allergenic potential of potato tubers. These data suggest, overall, a null impact of slcdi expression on tuber composition and substantial equivalence between comparator and SlCDI‐expressing tubers despite reported effects on leaf protein catabolism. They also illustrate the usefulness of proteomics as a tool to assess the authenticity of foods derived from novel‐generation transgenic plants.     

Comparison of airborne spore concentrations and fungal allergen content

The exposure to spores causing health effects is usually assessed by determining the concentration of viable spores per cubic meter of air (CFU/m³).Since allergens might also be present in dead spores or smaller particles, the objective of this study was to investigate the correlation between the viable spores of Alternaria and Cladosporium at different indoor and outdoor sites and the corresponding allergen concentration detected with a specially developed ELISA (Enzyme Linked Immunosorbent Assay). In outdoor air, the results show a strong correlation between the different sampling techniques applied for viable spores (Slit-Sampler and Multistage Liquid Impinger) and between the viable spores and the allergen concentrations detected in the liquid samples of the impingers. Indoors, the number of viable spores and the allergen concentration do not correlate and the allergen load is underestimated if colony counting methods are used. This revised version was published online in June 2006 with corrections to the Cover Date.     

Indoor air quality in schools: exposure to fungal allergens

This study examined indoor air quality within schools in Kansas City, Spokane, Santa Fe, and Orlando. Air sampling was undertaken with both Andersen Single Stage Samplers and Burkard Personal Air Samplers. The data show a wide range of indoor exposures ranging from less than 100 colony forming units (CFU/m³) for viable fungi and 100 spores/m³ for total spores in Spokane and Santa Fe to concentrations over 6000 CFU/m³ for viable fungi and 15 000 spores/m³ for total fungi in Orlando and Kansas City, respectively. In the majority of sites the indoor airspora reflected the outdoor taxa withCladosporium the most abundant genus identified; however, several indoor locations had elevated levels ofPenicillium andAspergillus indicating possible sources of indoor contamination. Airborne basidiospores and smut spores were also fairly abundant in the schools and were among the top five taxa identified. The data also indicated that the airborne concentrations vary significantly during the day and between classrooms within each school. Continued studies in schools are needed to fully assess both the exposure levels and the clinical significance to atopic children allergic to these spores.     

Purification and properties of ap-nitrobenzyl esterase fromBacillus subtilis

A procedure for purifying to homogeneity a microbially produced biocatalyst useful for deblocking intermediates in the manufacture of beta-lactam antibiotics is reported. In aqueous solution the purifiedp-nitrobenzyl (PNB) carboxy-esterase was soluble, monomeric (molecular weight: 54 000 by SDS-PAGE or by gel filtration) and exhibited an acidic pl, 4.1. The PNB carboxy-esterase catalyzed rapid ester hydrolysis for simple organic esters such as PNB-acetate, benzyl acetate and -naphthyl acetate and catalyzed deblocking (ester hydrolysis) of beta-lactam antibiotic PNB esters such as cephalexin-PNB and loracarbef-PNB. TheN-terminal amino acid sequence and the amino acid composition are reported. A serine residue is involved in ester hydrolysis: the PNB carboxy esterase was inhibited by phenylmethylsulfonyl fluoride and diethylp-nitrophenyl phosphate; one mole of diisopropyl fluorophosphate titration was required per mole of PNB carboxy-esterase for complete inhibition. When the [³H]-diisopropyl fluorophosphate-treated biocatalyst was digested with Lys C and the resulting peptides separated by HPLC, a single [³H]-labeled peptide was obtained; its amino acid sequence is reported. Inhibition of the PNB carboxy esterase by diethyl pyrocarbonate suggests that a histidinyl residue (or residues) is (are) also involved in the catalytic site of the esterase.Abbreviations used -ME -mercaptoethanol - Cf cefaclor - Cf nucleus-PNB - (6R, 7R) 7-amino-3-chloro-8-oxo-5-thia-1-azabicyclo[4.2.0]-oct-2-ene-2-carboxylic acid, (4-nitrophenyl)methyl ester - Cp cephalexin - Cp-PNB p-nitrobenzyl carboxy-ester of cephalexin - DEPC diethyl, pyrocarbonate - DFP diisopropyl fluorophosphate - DMSO dimethyl sulfoxide - DNP diethylp-nitrophenyl phosphate - EDTA ethylenediaminetetraacetic acid - EGTA ethylene, glycol-bis(aminoethyl ether) - N,N,NN tetracetic acid - Lc loracarbef - Lc-PNB p-nitrobenzyl carboxy-ester of loracarbef - Lc nucleus-PNB - (6R, 7S) 7-amino-3-chloro-8-oxo-1-azabicyclo[4.2.0]-oct-2-ene-2-carboxylic acid, (4-nitrophenyl)methyl ester - Lys C an endoproteinase specifically cleaving at C terminal lysine residues - MW_r relative molecular weight - PAGE polyacrylamide gel electrophoresis - PMSF phenylmethylsulfonylfluoride - PNB p-nitrobenzyl - PNBCE p-nitrobenzyl carboxy-esterase - SDS sodium dodecyl sulfate     

Optimization of electrophoresis for the identification of low molecular mass allergens in hazelnuts

Conventional electrophoresis techniques used to identify food allergens are insufficient to separate low molecular mass proteins and peptides. In this paper we performed three different methods which provided an extended resolving power for small proteins. Applying the improved techniques, we were able to separate hazelnut proteins into distinct bands below 10 kDa.