Andreas L. Lopata

Interaction of nanoparticles with proteins: relation to bio-reactivity of the nanoparticle

Interaction of nanoparticles with proteins is the basis of nanoparticle bio-reactivity. This interaction gives rise to the formation of a dynamic nanoparticle-protein corona. The protein corona may influence cellular uptake, inflammation, accumulation, degradation and clearance of the nanoparticles. Furthermore, the nanoparticle surface can induce conformational changes in adsorbed protein molecules which may affect the overall bio-reactivity of the nanoparticle. In depth understanding of such interactions can be directed towards generating bio-compatible nanomaterials with controlled surface characteristics in a biological environment. The main aim of this review is to summarise current knowledge on factors that influence nanoparticle-protein interactions and their implications on cellular uptake.

Occupational seafood allergy: a review

BACKGROUND Recent years have seen increased levels of production and consumption of seafood, leading to more frequent reporting of allergic reactions in occupational and domestic settings. This review focuses on occupational allergy in the fishing and seafood processing industry. REVIEW Workers involved in either manual or automated processing of crabs, prawns, mussels, fish, and fishmeal production are commonly exposed to various constituents of seafood. Aerosolisation of seafood and cooking fluid during processing are potential occupational situations that could result in sensitisation through inhalation. There is great variability of aerosol exposure within and among various jobs with reported allergen concentrations ranging from 0.001 to 5.061(μg/m3). Occupational dermal exposure occurs as a result of unprotected handling of seafood and its byproducts. Occupational allergies have been reported in workers exposed to arthropods (crustaceans), molluscs, pisces (bony fish) and other agents derived from seafood. The prevalence of occupational asthma ranges from 7% to 36%, and for occupational protein contact dermatitis, from 3% to 11%. These health outcomes are mainly due to high molecular weight proteins in seafood causing an IgE mediated response. Cross reactivity between various species within a major seafood grouping also occurs. Limited evidence from dose-response relations indicate that development of symptoms is related to duration or intensity of exposure. The evidence for atopy as a risk factor for occupational sensitisation and asthma is supportive, whereas evidence for cigarette smoking is limited. Disruption of the intact skin barrier seems to be an important added risk factor for occupational protein contact dermatitis. CONCLUSION The range of allergic disease associated with occupational exposure to crab is well characterised, whereas for other seafood agents the evidence is somewhat limited. There is a need for further epidemiological studies to better characterise this risk. More detailed characterisation of specific protein antigens in aerosols and associated establishment of dose-response relations for acute and chronic exposure to seafood; the respective roles of skin contact and inhalational exposure in allergic sensitisation and cross reactivity; and the contribution of host associated factors in the development of occupational seafood allergies are important areas for future research.

New insights into seafood allergy

Purpose of reviewSeafood plays an important role in human nutrition worldwide, sustained by international trade of a variety of new seafood products. Increased production and consumption have resulted in more frequent reports of adverse reactions, highlighting the need for more specific diagnosis and treatment of seafood allergy. This review discusses recent literature in this field. Recent findingsThe most recent prevalence data from Asia highlight seafood as a significant sensitizer in up to 40% of children and 33% of adults. Furthermore, the demonstration of species-specific sensitization to salt-water and fresh-water prawns and processed prawn extract should improve diagnosis. Studies on humans demonstrated for the first time that biologically active fish allergens can be detected in serum samples as early as 10 min after ingestion. These studies highlight that minute amounts of ingested seafood allergens can quickly trigger allergic symptoms; also, inhaled airborne allergens seem to induce sensitization and reactions. In the past 2 years, over 10 additional seafood allergens have been characterized. Allergen-specific detection assays in food products are available for crustacean tropomyosin; however, many specific mollusk and some fish allergens are not readily identified. SummaryAlthough cross-reactivity between crustacean and mollusks as well as mites is demonstrated, the often poor correlation of IgE reactivity and clinical symptoms calls for more detailed investigations. The recent development of hypoallergenic parvalbumin from carp could form the basis for safer vaccination products for treatment of fish allergy. Molecular characterization of more universal marker allergens for the three major seafood groups will improve current component-resolved clinical diagnosis and have a significant impact on the management of allergic patients, on food labeling and on future immunotherapy for seafood allergy.

EAACI Molecular Allergology User's Guide

The availability of allergen molecules (‘components’) from several protein families has advanced our understanding of immunoglobulin E (IgE)‐mediated responses and enabled ‘component‐resolved diagnosis’ (CRD). The European Academy of Allergy and Clinical Immunology (EAACI) Molecular Allergology Users Guide (MAUG) provides comprehensive information on important allergens and describes the diagnostic options using CRD. Part A of the EAACI MAUG introduces allergen molecules, families, composition of extracts, databases, and diagnostic IgE, skin, and basophil tests. Singleplex and multiplex IgE assays with components improve both sensitivity for low‐abundance allergens and analytical specificity; IgE to individual allergens can yield information on clinical risks and distinguish cross‐reactivity from true primary sensitization. Part B discusses the clinical and molecular aspects of IgE‐mediated allergies to foods (including nuts, seeds, legumes, fruits, vegetables, cereal grains, milk, egg, meat, fish, and shellfish), inhalants (pollen, mold spores, mites, and animal dander), and Hymenoptera venom. Diagnostic algorithms and short case histories provide useful information for the clinical workup of allergic individuals targeted for CRD. Part C covers protein families containing ubiquitous, highly cross‐reactive panallergens from plant (lipid transfer proteins, polcalcins, PR‐10, profilins) and animal sources (lipocalins, parvalbumins, serum albumins, tropomyosins) and explains their diagnostic and clinical utility. Part D lists 100 important allergen molecules. In conclusion, IgE‐mediated reactions and allergic diseases, including allergic rhinoconjunctivitis, asthma, food reactions, and insect sting reactions, are discussed from a novel molecular perspective. The EAACI MAUG documents the rapid progression of molecular allergology from basic research to its integration into clinical practice, a quantum leap in the management of allergic patients.

Shellfish allergy: Shellfish allergy

Seafood plays an important role in human nutrition and health. The growing international trade in seafood species and products has added to the popularity and frequency of consumption of a variety of seafood products across many countries. This increased production and consumption of seafood has been accompanied by more frequent reports of adverse health problems among consumers as well as processors of seafood. Adverse reactions to seafood are often generated by contaminants but can also be mediated by the immune system and cause allergies. These reactions can result from exposure to the seafood itself or various non‐seafood components in the product. Non‐immunological reactions to seafood can be triggered by contaminants such as parasites, bacteria, viruses, marine toxins and biogenic amines. Ingredients added during processing and canning of seafood can also cause adverse reactions. Importantly all these substances are able to trigger symptoms which are similar to true allergic reactions, which are mediated by antibodies produced by the immune system against specific allergens. Allergic reactions to ‘shellfish’, which comprises the groups of crustaceans and molluscs, can generate clinical symptoms ranging from mild urticaria and oral allergy syndrome to life‐threatening anaphylactic reactions. The prevalence of crustacean allergy seems to vary largely between geographical locations, most probably as a result of the availability of seafood. The major shellfish allergen is tropomyosin, although other allergens may play an important part in allergenicity such as arginine kinase and myosin light chain. Current observations regard tropomyosin to be the major allergen responsible for molecular and clinical cross‐reactivity between crustaceans and molluscs, but also to other inhaled invertebrates such as house dust mites and insects. Future research on the molecular structure of tropomyosins with a focus on the immunological and particularly clinical cross‐reactivity will improve diagnosis and management of this potentially life‐threatening allergy and is essential for future immunotherapy.

Occupational Allergy and Asthma Among Salt Water Fish Processing Workers

BACKGROUND Fish processing is a common economic activity in Southern Africa. The aim of this study was to determine the prevalence and host determinants of allergic symptoms, allergic sensitization, bronchial hyper-responsiveness and asthma among workers processing saltwater fish. METHODS A cross-sectional study was conducted on 594 currently employed workers in two processing plants involved in pilchard canning and fishmeal processing. A modified European Community Respiratory Health Survey (ECRHS) questionnaire was used. Skin prick tests (SPT) used extracts of common airborne allergens, fresh fish (pilchard, anchovy, maasbanker, mackerel, red eye) and fishmeal. Spirometry and methacholine challenge tests (MCTs; tidal breathing method) used ATS guidelines. RESULTS Work-related ocular-nasal symptoms (26%) were more common than asthma symptoms (16%). The prevalence of atopy was 36%, while 7% were sensitized to fish species and 26% had NSBH (PC(20) < or = 8 mg/ml or > or =12% increase in FEV(1) post-bronchodilator). The prevalence of probable occupational asthma was 1.8% and fish allergic rhino-conjunctivitis 2.6%. Women were more likely to report work-related asthma symptoms (OR = 1.94) and have NSBH (OR = 3.09), while men were more likely to be sensitized to fish (OR = 2.06) and have airway obstruction (OR = 4.17). Atopy (OR = 3.16) and current smoking (OR = 2.37), but not habitual seafood consumption were associated with sensitization to fish. CONCLUSIONS Based on comparison with previous published studies, the prevalence of occupational asthma to salt water fish is lower than due to shellfish. The gendered distribution of work and exposures in fish processing operations together with atopy and cigarette smoking are important determinants of occupational allergy and asthma.

Biomolecular characterization of allergenic proteins in snow crab (Chionoecetes opilio) and de novo sequencing of the second allergen arginine kinase using tandem mass spectrometry

Snow crab (Chionoecetes opilio) proteins have been recognized as an important source of both food and occupational allergens. While snow crab causes a significant occupational allergy, only one novel allergen has recently been fully characterized. The muscle proteins from snow crab legs were profiled by SDS-PAGE. Several of these proteins were characterized using tandem mass spectrometry. Five proteins were identified; sarcoplasmic Ca-binding (20kDa), arginine kinase (40), troponin (23kDa) and α-actine (42kDa) and smooth endoplasmic reticulum Ca(2+)ATPase (113kDa). Immunoblotting using serum of sixteen allergic patients resulted in strong reactivity with the 40-kDa protein in seven patients (43%). This protein was purified by chromatography and subsequently de novo sequenced using matrix assisted laser desorption ionization and electrospray tandem mass spectrometry. We identified a second important allergen, arginine kinase, in snow crab, designated Chi o 3. Based on identity and homology analysis, using bioinformatics tools, a signature peptide was identified as a chemical surrogate for arginine kinase. The suitability of this signature peptide was tested for analytically representing the arginine kinase, by performing a multi-reaction monitoring tandem mass spectrometry approach on actual air filter samples collected from a simulated crab processing plant.

Impact of heat processing on the detection of the major shellfish allergen tropomyosin in crustaceans and molluscs using specific monoclonal antibodies.

The major heat-stable shellfish allergen, tropomyosin, demonstrates immunological cross-reactivity, making specific differentiation of crustaceans and molluscs for food labelling very difficult. The aim of this study was to evaluate the application of allergen-specific monoclonal antibodies in differential detection of shellfish-derived tropomyosin in 11 crustacean and 7 mollusc species, and to study the impact of heating on its detection. Cross-reactive tropomyosin was detected in all crustacean species, with partial detection in molluscs: mussels, scallops and snails but none in oyster, octopus and squid. Furthermore, we have demonstrated that heating of shellfish has a profound effect on tropomyosin detection. This was evident by the enhanced recognition of multiple tropomyosin variants in the analysed shellfish species. Specific monoclonal antibodies, targetting the N-terminal region of tropomyosin, must therefore be developed to differentiate tropomyosins in crustaceans and molluscs. This can help in correct food labelling practices and thus protection of consumers.

Characteristics of hypersensitivity reactions and identification of a unique 49 kd IgE-binding protein (Hal-m-1) in abalone (Haliotis midae)

BACKGROUND There is a paucity of published data on the clinical presentation and the nature of the allergens involved in hypersensitivity to mollusks. This study reports the clinical and immunologic findings in 38 patients with reported immediate and delayed adverse reactions to abalone (Haliotis midae, Class Gastropoda). METHODS Patients were recruited as part of a South African seafood allergy survey. Allergic symptoms were assessed by a self-administered questionnaire. A total of 38 patients with abalone sensitivity were recruited. Specific IgE responses to abalone and other mollusks were studied by using RAST and inhibition ELISAs. Skin prick tests and lymphocyte proliferation assays were also performed on several of the subjects. Allergenic components of Haliotis midae were identified with Western blotting. RESULTS Twenty five of the 38 patients in the study were first seen with immediate symptoms, and 13 had delayed reactions. Seventeen of the sera tested were RAST positive. Skin prick tests responses with abalone extract were positive in all subjects with positive RAST responses (n = 8) and in 6 of 13 subjects with negative RAST responses. Five of the subjects with positive RAST responses had positive results on Western blotting and demonstrated binding to two major allergens with molecular weights of 38 and 49 kd. The 49 kd IgE-binding protein has been designated as Hal-m-1. CONCLUSIONS Abalone allergens are heat-stable proteins with molecular weights of 38 and 49 kd, later designated as Hal-m-1 according to International Union of Immunological Societies allergen nomenclature regulation. Our studies indicate a clear clinical and immunologic heterogeneity in patients reactive to abalone.

Biological reactivity of zinc oxide nanoparticles with mammalian test systems: an overview

Zinc oxide nanoparticles (ZnO NPs) have useful physicochemical advantages, and are used extensively. This has raised concerns regarding their potential toxicity. ZnO NP attributes that contribute to cytotoxicity and immune reactivity, however, seem to vary across literature considerably. Largely, dissolution and generation of reactive oxygen species appear to be the most commonly reported paradigms. Moreover, ZnO NP size and shape may also contribute toward their overall nano-bio interactions. Analysis is further complicated by factors such as adsorption of proteins on the NP surface, which may influence their bioreactivity. The main aim of this review is to give a systematic overview of the postulates explaining cytotoxic, inflammatory and genotoxic effects of ZnO NPs when exposed to different types of cells in vitro and in vivo.